Glutamine, Glutamate, and Aspartate Improves Morphology and Energy Production of Small Intestine in Piglets with Different Energy Levels Diets

Glutamine, Glutamate, and Aspartate Improves Morphology and Energy Production of Small Intestine in Piglets with Different Energy Levels Diets | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Glutamine, Glutamate, and Aspartate Improves Morphology and Energy Production of Small Intestine in Piglets with Different Energy Levels Diets Jing Wang, Nan Wang, Ming Qi, Jianjun Li, Bie Tan, Yulong Yin This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-150010/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Background: Weaning-stress may cause reduced energy intake for maintenance of mucosal structure. Glutamine (Gln), glutamate (Glu), and aspartate (Asp) are major energy sources for small intestine. This study investigated whether Gln, Glu, and Asp improve the intestinal morphology via regulating the energy metabolism in weaning-piglet. A total of 198 weaned-piglets were assigned to the following treatments: Control (Basal diet + 1.59% L-Alanine); T1 (Basal diet + 1% L-Gln + 0.5% L-Glu + 0.1% L-Asp); T2 (Low energy diet + 1% L-Gln + 0.5% L-Glu + 0.1% L-Asp). Jejunum and ileum were obtained on d-5 or d-21 post-weaning. Results: Improved growth performance on d-21 post-weaning were observed in T1 treatment. Both T1 and T2 treatments improved small intestinal morphology by increasing villus height, goblet cell number and decreasing crypt depth on d 5 or d-21 post-weaning. Gln, Glu, and Asp could restore the small intestinal energy homeostasis via replenishing the Krebs’ cycle and down-regulate the AMPK pathway. However, when piglets fed by a low energy diet, Gln, Glu, and Asp are not sufficient to maintain the intestinal energy balance on d-5 post-weaning so that the AMPK, beta-oxidation, and mitochondrial biogenesis are activated to meet the high energy demand of enterocytes. Conclusion: These data indicated that Gln, Glu, and Asp could restore the energy homeostasis of intestinal mucosa of piglets. And the mucosal energy metabolism showed different responses to the intervention of Gln, Glu, and Asp in piglets with a low energy diet between d5 and d21 post-weaning. These findings provide new information on the nutritional intervention for the insufficient energy intake in weaning-piglet. Animal Science Biotechnology and Bioengineering Nutrition & Dietetics glutamine glutamate aspartate small intestine energy metabolism Figures Figure 1 Figure 2 Figure 3 Figure 4 Background At weaning, young pigs are subjected to removal from sow and littermates and are transported to a new environment, as well as their diet is abrupt changed from sow’s milk to solid diet, which cause low voluntary food intake and associated poor intestinal barrier structure and function [1, 2]. Undoubtedly, the weaning pigs are not eating enough food and their intestinal mucosa is failing to digest and absorb sufficient nutrients to cover their energy requirement for maintenance [3], which are major limitations to improve the growth of piglet after-weaning. Therefore, improving the energy intake from exogenous nutrition is rationally served as a promising approach to protect piglets from weaning stress. The intestine of piglet has notably high energy demand due to the repaid renewal of epithelium within few days [4, 5]. Hence, the epithelial cells of gastrointestinal (GI) tract require intense anabolic metabolism. It has been demonstrated that GI tract represents approximately 5% of body weight, whereas it is responsible for about 20% of whole body O 2 consumption [6]. Glucose and lipid are major sources for the supply and storage of energy in cells [7]. AMP-activated protein kinase (AMPK) is the master regulator of energy metabolism [6]. AMPK increases adenosine triphosphate (ATP) levels by promoting glucose and lipids breakdown and inhibiting their synthesis and storage [7]. Glycolysis, Krebs’ cycle, and fatty acids beta-oxidation are the main catabolic processes for glucose and lipids [8], respectively. Although all animals obtain their biological energy from the cell-specific oxidation of fatty acids, glucose, and amino acids in diets [9], for gastrointestinal tract, glutamine (Gln), glutamate (Glu), and aspartate (Asp) are the main energy sources to maintain gut integrity and function [10, 11]. Meanwhile, numerous studies have reported that Gln, Glu, and Asp has profound impacts on the intestinal nutrition and health. Gln plays multiple roles in regulating intestinal protein turnover [12], gene expression [13], cell proliferation [14] and immune function [15]. Asp could improve growth performance of weanling piglets [16], and attenuate the intestinal injury induced by Escherichia coli lipopolysaccharide (LPS) [17]. Nevertheless, a typical corn- and soybean meal-based diet cannot provide sufficient amounts of Asp, Glu, and Gln for protein synthesis in post-weaning pigs [18]. In the present study, we hypothesize that Gln, Glu, and Asp could improve the small intestinal structure of weanling piglets via regulating the enterocyte energy metabolism. The effects of supplementation of Gln, Glu, and Asp on the intestinal morphology, energy metabolites, AMPK pathway phosphorylation were determined in weaning piglets on d 5 and d 21 post-weaning. Methods All animals used in this study were humanely managed according to the Chinese Guidelines for Animal Welfare. The experimental protocol was approved by the Animal Care and Use Committee of the Chinese Academy of Sciences (Beijing, China). Animals and experimental design A total of 198 piglets (Duroc ×Landrace × Large Yorkshire) weaned at 21-d of age were assigned to 18 pens based on their body weight (BW). There were 11 piglets per pen and 6 pens per treatment. The treatment groups include: i) Control (Basal diet + 1.59% L-Alanine (Ala); iso-nitrogenous control); ii) T1 (Basal diet + 1% L-Gln + 0.5% L-Glu + 0.1% L-Asp); iii) T2 (Low energy diet + 1% L-Gln + 0.5% L-Glu + 0.1% L-Asp) (Table 1). The basal diets were formulated to meet the nutrient requirements recommended by the National Research Council (2012). A low energy level diet was obtained by removing the soybean oil, glucose, and sucrose. The dose was based on the growth performance of piglets in the preliminary experiment. All piglets were housed in an environmentally well controlled nursery facility with slatted plastic flooring and a mechanical ventilation system, and had free access to drink water. One piglet closed to the average body weight from each pen was slaughtered on day 5 (w5d) and 21 (w21d) post-weaning. After being stunned electrically, piglets were sacrificed and the jejunum and ileum was dissected and rinsed thoroughly with ice-cold physiological saline. The middle segments of the jejunum (2 cm) and ileum (2 cm) were cut and fixed in 2.5% glutaraldehyde or 4% formaldehyde for morphological and immunohistochemical analysis. Samples of the jejunal and ileal mucosa were scraped, immediately snap frozen in liquid nitrogen and stored at -80 °C for further analysis. Intestinal morphology The segments of the jejunum and ileum fixed in 4 % formaldehyde were used to determine morphology using hematoxylin-eosin staining. After dehydration, embedding, sectioning, and staining, images were acquired at various magnifications with computer-assisted microscopy (Micro-metrics TM; Nikon ECLIPSE E200, Tokyo, Japan). Villous height, crypt depth, goblet cell and lymphocyte counts were measured by Image-Pro Plus software, Version 6.0 on images at 200- or 400-fold magnification in five randomly selected fields, respectively [19]. Segments of the jejunum and ileum at 150-fold magnification were also designated for analysis by scanning electron microscopy as described by German [20] and Liu et al. [21]. Briefly, tissue segments were fixed with 2.5 % glutaraldehyde for 2 h at 4 °C, and rinsed 3×10 min in PBS at 4 °C. The tissues were then postfixed in 1 % osmium tetroxide for 12 h at 4 °C, and rinsed 3×10 min in PBS at 4 °C. After samples were mounted onto stubs by means of quick-drying silver paint, the tissues were coated with gold-palladium and examined by a JEOL JSM-6360LV scanning electron microscope at 25 KV. The apparent characteristics of microvillus were observed and described. After samples were mounted onto stubs by means of quick-drying silver paint, the tissues were coated with gold-palladium and examined by a JEOL SU8010 scanning electron microscope at 1.0 KV. The apparent characteristics of microvillus were observed and described. Metabolomic profiling The ileal mucosal metabolomic profile was analyzed by liquid chromatography (LC)- mass spectrometry (MS)/MS. Briefly, 100 mg ileal mucosa was homogenized in 200 µL ddH 2 O, and mixed with 800 µL methanol/acetonitrile (1:1, v/v) and sonicated in ice water-bath for 30 min. After incubation in -20 °C for 1 h, the mixture was centrifuged at 14,000 g for 15 min at 4 °C. The supernatant was dried by speedvac, resuspended in 100 µL methanol/ddH 2 O (1:1, v/v) for LC-MS/MS analysis. An aliquot from each sample was pooled to create quality control (QC) samples that were used to evaluate the internal standards and instrument performance. The ultra-high-performance LC (UPLC; Agilent 1290 Infinity LC, USA) was performed on a 2.1 x 100 mm ACQUITY UPLC BEH Amide column (internal diameter 1.7 μm; Waters, USA). The column was warmed to 45 °C before use. The mobile phase for UPLC analysis consisted of two solutions: (A) 15 mM ammonium acetate/H 2 O and (B) acetonitrile. The MS/MS (5500 QTRAP, AB SCIEX, USA) spectra was set as follows: source temperature 450 °C, ion Source Gas 1: 45, Ion Source Gas 2: 45, Curtain gas: 30. ionSapary Voltage Floating-4500 V. Real-Time Quantitative Reverse Transcriptase PCR Expressions of pyruvate dehydrogenase kinase ( Pdk4 ), phosphoenolpyruvate carboxykinase 1 ( Pck1 ), succinate dehydrogenase 1 ( Sdh1 ), mitochondrial uncoupling protein 2 ( Ucp2 ), peroxisome proliferator-activated receptor alpha 1 ( Ppar α 1 ), peroxisomal acyl-coenzyme A oxidase 1 ( Acox1 ), carnitine palmitoyltransferase 1A ( Cpt1a ), acetyl-CoA carboxylase ( Acc ), Ampk , Sirtuin 1 ( Sirt1 ), peroxisome proliferator-activated receptor gamma coactivator 1-α ( Pgc1α ), mitochondrial transcription factor A ( Tfam ), and nuclear respiratory factor 1 ( Nrf1 ) mRNA in jejunal and ileal mucosa were determined by real-time quantitative reverse transcriptase PCR (real-time qRT-PCR) as described previously [22]. Primers were designed with Primer 5.0 (PREMIER Biosoft International, Palo Alto, CA) according to the gene sequence of the pig to produce an amplification product (Table S1). The comparative threshold cycle (Ct) value method (2 -ΔΔCt ) was employed to quantitate expression levels for target genes relative to those for the β-Actin. Data were expressed as the relative values to those of control piglets. Western blotting Jejunal and ileal mucosa samples were homogenized, and protein concentrations were measured using the bicinchoninic acid assay method with BSA as standard (Beyotime Institute of Biotechnology, Shanghai, China). All samples were adjusted to an equal concentration (70 µg protein). The supernatant fluid (containing tissue proteins) was diluted with 5 × sodium dodecyl sulfate (SDS) sample buffer and heated in boiling water for 5 min and cooled down for Western blot analysis [23]. The following first antibodies were used for protein quantification: AMPKα (1:1,000; Cell Signaling Technology, USA), phosphorylated AMPKα (p-AMPK (Thr175); 1:1,000; Cell Signaling Technology, USA), ACC (1:1,000; Cell Signaling Technology, US), phosphorylated ACC (p-ACC (Ser79); 1:1,000; Cell Signaling Technology, USA), SirT1 (1:1,000; Cell Signaling Technology, USA), PGC1α (1:1,000; Abcam, UK) and β-actin (1:1000; Cell Signaling Technology, USA) as well as secondary antibodies (horseradish peroxidase-conjugated goat anti-rabbit IgG; 1:5000; Boster Biological Technology, Wuhan, China). The images were detected by chemiluminescence (Millipore, Billerica, MA). Each Western blot was subjected to multiple exposures to ensure that the chemiluminescence signals were linear. Western blots were quantified by measuring the intensity of correctly sized bands using software (Alpha Imager 2200 Software; Alpha Innotech Corporation, San Leandro, CA, USA) and protein measurement was normalized to β-actin. Statistical analysis The growth performance was performed by one-way ANOVA using SPSS software 19.0 (SPSS Inc., Chicago, IL). The differences among treatments were evaluated using Tukey’s test. The intestinal morphology, metabolomic profiles, gene and protein expression were analyzed using the general linear models’ procedure, with diet treatments, days after weaning, and the interaction between treatment and day as the main effects. The results in the tables were presented as main effects. When no significant interaction between treatment and day was found, the differences among treatments or days were evaluated using Tukey HSD test (showed in Table 3, 4, 5, 6). When significant interaction between treatment and day was found, a simple main effect analysis would be performed and the differences among 3 treatments across 2 levels of days post-weaning were evaluated using Bonferroni test (showed in Figure 1, 2, 3). The developmental changes of the intestine were not the main outcome, so the differences between d 5 and d 21 post-weaning across 3 levels of treatments were not showed. Differences were declared as significant at P < 0.05. Results Growth performance Initial body weight (BW), final BW, average daily gain (ADG), average daily feed intake (ADFI), and the ratio of average daily feed intake to average daily gain (F / G) were showed in Table 2. The initial BW was similar among treatments ( P = 0.959). On d 5 post-weaning, there were no differences on BW ( P = 0.952), ADG ( P = 0.782), ADFI ( P = 0.127) and F/G ( P = 0.240) among different treatments. On d 21 post-weaning, basal dietary supplementation with Gln, Glu, Asp (T1) improved the final BW and ADG of piglets in comparison to those in control group ( P < 0.05). However, the digestible energy intakes of weanling piglets neither on d 5 ( P = 0.190) nor d 21 ( P = 0.709) post-weaning were not significantly different. Jejunal and ileal morphology In Figure 1A and B, the scanning electron microscopy showed that the villi in the jejunum and ileum on d 5 post-weaning were thin and sparse, and then grew shorter, stouter and denser on d 21 post-weaning. In the jejunum, the villi heights among control, T1, and T2 groups were not showed significantly different on w 5 d, but there was an increase in villi density and villi-fold in T1 and T2 group on w 21 d. In the ileum, the increased villi densities were observed in control and T1 groups in comparison to T2 group on w 5 d, whereas villi in T1 and T2 group were denser and stouter than villi in control group on w 21 d. The main effects of diet treatments and days after weaning on parameters from the intestinal hematoxylin-eosin staining were showed in Table 3. When significant interactions between treatments and days after weaning were obtained, differences among control, T1 and T2 on d 5 or d 21 post-weaning were showed in Figure 1C. In Table 3, the number of goblet cells in the jejunum of T2 group was higher than these in T1 group ( P < 0.05). There were interactions ( P < 0.05) between treatments and days on the jejunal villus height, ileal villus height and crypt depth, which means days after weaning influenced the treatments’ efficacy. In Figure 1C, a low energy diet supplemented with Glu, Gln, and Asp increased ( P < 0.05) the villus height of jejunum on d 21 post-weaning and decreased ( P < 0.05) the crypt depth of ileum on d 5 post-weaning as compared to the control group, whereas it had lower ( P < 0.05) ileal villus height than the control and T1 groups on d 5 post-weaning. A basal diet supplemented with Gln, Glu, and Asp increased the villus height of ileum on d 21 post-weaning compared to the control group. Metabolomic profiling of jejunal mucosa of weanling piglets The jejunal mucosa energy metabolite fingerprints were presented in Figure 2A and Figure 2B. Piglets on d 5 and d 21 postweaning could be readily differentiated by the heat map of their jejunal mucosa energy metabolites (Figure 2A). Piglets on d 5 post-weaning showed more abundances of dihydroxyacetone phosphate (DHAP), isocitrate, aconitate, citrate, guanosine 5 monophosphate (GMP), fructose 6 phosphate (F-6-P), and glucose-6-phosphate (G-6-P) in jejunal mucosa, while piglets on d 21 post-weaning showed more abundances of fumarate, malate, flavin mononucleotide (FMN), cyclic- adenosine 5 monophosphate (c-AMP), AMP, nicotinamide adenine dinucleotide phosphate hydrogen (NADPH), nicotinamide adenine dinucleotide phosphate (NADP + ), nicotinamide adenine dinucleotide (NAD + ), adenosine 5 diphosphate (ADP), and thiamine pyrophosphate(TPP) in jejunal mucosa. The first two components of a principal component analysis (PCA) showed that the jejunal mucosal energy metabolites of control and T1 treatments on d 5 post-weaning were comparable, while T2 treatment on d 5 post-weaning was more dispersive and different from control and T1 treatments. On d 21 post-weaning, energy metabolites in T1 and T2 treatments were comparable, which were different from these in the control group. The main effects of diet treatments and days after weaning on metabolites’ levels of jejunal mucosa were showed in Table S2. When significant interactions between treatments and days after weaning were found, differences among control, T1 and T2 on d 5 or d 21 post-weaning were showed in Figure 2C. In Table S2, supplementation with Gln, Glu, and Asp in both basal and low energy diets increased ( P < 0.05) the succinate level but decreased ( P < 0.05) the phosphoenolpyruvate level in jejunal mucosa of weanling piglets. Days post-weaning had influences on the diet treatments’ efficacy ( P interaction < 0.05). As shown in Figure 2C, a basal diet supplemented with Gln, Glu, and Asp (T1) increased ( P < 0.05) the pyruvate, α-ketoglutarate (AKG), isocitrate, and fructose 1,6 phosphate (F-1,6-P) levels but decreased ( P < 0.05) the NADPH level on d 5 post-weaning compared to control group. A low energy diet supplemented with Gln, Glu, and Asp (T2) increased ( P < 0.05) the pyruvate, AKG, isocitrate and GMP levels compared to the control group and increased ( P < 0.05) DHAP, NADPH, and AMP levels as compared to control and T1 groups but reduced ( P < 0.05) 3-phosphoglycerate, F-6-P, and G-6-P levels on d 5 post-weaning. On d 21 post-weaning, T1 and T2 treatments showed higher ( P < 0.05) oxaloacetate level than the control group. Expression of gene involved in energy metabolism of jejunal and ileal mucosa of piglets The gene expressions of several key enzymes of energy metabolism were presented in Table S3 and Figure 2D. The main effect in Table S3 showed T1 treatment increased ( P < 0.05) the ileal Ucp2 mRNA level compared to the control group but decreased ( P < 0.05) the ileal Cpt1a mRNA level compared to control and T2 groups. T2 treatment increased ( P < 0.05) the jejunal Acox1 and ileal Cpt1a mRNA levels in comparison with the control group. Days post-weaning had influences on the diet treatments’ efficacy ( P interaction < 0.05; Figure 2D). On d 5 post-weaning, a low energy diet supplemented with Gln, Glu, and Asp had lower ( P < 0.05) Pdk4 and Pck1 mRNA abundances than control or T1 groups. On d 21 post-weaning, a basal diet supplemented with Gln, Glu, and Asp enhanced ( P < 0.05) the Sdh1 mRNA abundance in comparison to these in the control group. A low energy diet supplemented with Gln, Glu, and Asp increased ( P < 0.05) the Pck1 and Sdh1 mRNA abundances as compared to T1 or control groups, respectively. mRNA and protein abundances of AMP-activated protein kinase signaling pathway in jejunal and ileal mucosa of piglets The relative mRNA and protein abundances of AMPK pathway in the jejunal and ileal mucosa were presented in Table S4, S5 and Figure 3, respectively. The main effects on AMPK pathway gene expressions in Table S4 showed a basal diet supplemented with Gln, Glu and Asp (T1) lowered ( P < 0.05) jejunal Pgc1α , Tfam , and Nrf1 and ileal Sirt1 , Pgc1α mRNA levels compared to control and T2 group, as well as ileal Tfam mRNA level compared to the T2 group. The main effects on AMPK pathway protein abundances in Table S5 showed T1 treatment enhanced ( P 0.05) but had influenced the treatments’ efficacy on AMPK pathway protein abundances ( P interaction < 0.05). As shown in Figure 3, a basal diet supplemented with Gln, Glu and Asp (T1) enhanced ( P < 0.05) the jejunal AMPKα protein abundance compared to the control group but reduced ( P < 0.05) the ileal p-AMPKα and PGC1α protein abundances as compared to control and T2 groups, respectively, on d 5 post-weaning. A low energy diet supplemented with Gln, Glu and Asp (T2) reduced ( P < 0.05) the ileal AMPKα protein abundance compared to control and T1 groups, while increased ( P < 0.05) jejunal and ileal SirT1 and PGC1α, as well as jejunal ACC and p-ACC protein abundances compared to control or T1 groups on d 5 post-weaning. On d 21 post-weaning, T1 and T2 treatments lowered ( P < 0.05) the jejunal ACC, ileal AMPKα and PGC1α, as well as jejunal and ileal p-AMPKα, p-ACC, and SirT1 protein abundances as compared to the control groups. Meanwhile, T1 treatment showed lower ( P < 0.05) jejunal and ileal PGC1α and ileal SirT1 protein abundances than these in the T2 group. Discussion The mammalian intestine plays a key role in nutrients intake and has a high rate of energy expenditure because the digestion and absorption processes are highly dependent on energy [24, 25].Weaning results in a growth check caused by a period of low feed intake, which also reduces the energy intake of intestine of piglets [26, 27]. Successful adaptation to these changes requires profound morphological and energy metabolic adjustments of the gastrointestinal tract [28]. Gln, Glu and Asp were used to help the faster restitution of mucosal barrier after stress injury, and to improve the intestinal energy metabolism of piglets with different energy levels diets in this study. The most serious damages in piglets during weaning-stress occurs on d 2 to d 5 post-weaning [25, 29], and is usually associated with a dramatic reduction of feed intake resulting in marked changes in the growth pattern on body weight [28]. In the present study, supplementation with 1 % Gln, 0.5 % Glu, and 0.1 % Asp in a normal energy level diet enhanced the BW and ADG on d 21 post-weaning. The beneficial effect of Gln, Glu and Asp on improving the growth-suppression induced by weaning stress in our study was consistent with previous researches, which have shown that Glu and Asp exert positive effects on growth performance of animals in healthy and oxidative-stress conditions [30, 31] [32, 33]. Although low energy diet did not significantly increase the ADFI of weanling piglets, the digestive energy intake in T2 group was comparable with these in control and T1 groups. Piglets in low energy diet treatment increased their satiety through increasing food intake. In addition to poor growth performance, the growth-check induced by weaning-stress is accompanied by impaired mucosal integrity, villous atrophy, an increased rate of cell loss, or a decreased rate of crypt-cell production [34, 35]. Supplementation with Gln, Glu, and Asp in normal energy diet increased ileal villus height on d 21 post-weaning. The major reasons for villus atrophy are deficient enteral nutrition and reduced energy intake independent of diet type [36, 37]. We found that a low energy diet supplemented with Gln, Glu, and Asp could alleviate impaired intestinal structure by raising jejunal villus height and goblet cells number or reducing ileal crypt depth on either d 5 or d 21 post-weaning. The critical roles of these three amino acids in intestine have been reported in many well-designed studies, for example, Gln could increase the villus length of jejunum [38], and Asp could alleviate the intestinal barrier damage [32, 39, 40]. The results in our current study indicated that Gln, Glu, and Asp could improve the intestinal barrier structure of weanling piglets fed either a low energy diet or a normal energy diet, which led us to further explore the mechanism of these three amino acids regulating energy homeostasis in the small intestine. ATP production and utilization are very active in pig enterocytes. The relative abundances of intermediate metabolites involved in the energy metabolism of the small intestine of piglets on d 5 and d 21 post-weaning were not comparable, and the regulation of Gln, Glu, and Asp on energy metabolites varied between d 5 and d 21 post-weaning. It may indicate that the energy metabolism in the small intestine changes during the weaning period [41], and the pattern of metabolism has different responses to amino acids. It is well established that dietary Gln, Glu, and Asp plus arterial Gln provide up to 80 % of ATP to the small intestine mucosa in mammals [11, 42, 43]. After these three amino acids are absorbed into enterocytes, they could be metabolized and oxidized via the Krebs’ cycle to water and CO2 yielding ATP [44]. The oxidative steps of Gln and Glu in pig enterocytes are similar. Gln firstly enter the mitochondrial in order to be degraded to Glu and ammonia by the mitochondrial glutaminase [45]. Glu could be transaminated to produce AKG, which is the key molecular of the Krebs’ cycle [46]. Asp replenished the Krebs cycle by being converted to oxaloacetate [44]. In the present study, supplementation with Gln, Glu, and Asp either in a regular energy diet or a low energy diet could replenish the jejunal Krebs’ cycle, manifested as increased pyruvate, oxaloacetate, isocitrate, AKG, and succinate levels on d 5 or d 21 post-weaning. Meanwhile, supplementation with Gln, Glu and Asp in either regular energy diet or low energy diet enhanced Sdh1 mRNA abundance on d 21 post-weaning, whereas low energy diet supplemented with Gln, Glu, and Asp declined Pdk4 mRNA abundance on d 5 post-weaning. SDH1 is part of Krebs’ cycle and oxidizes succinate to fumarate [47]. The higher Sdh1 expression was consistent with the higher succinate level in the jejunal mucosa. Sdh1 is responsible to promote the oxidation of succinate and prevents the accumulation of succinate in the mitochondria [48].PDK4 is a regulator of pyruvate dehydrogenase (PDH), as it could inactivate PDH by phosphorylation [49]. PDH represents a cornerstone in cellular energy metabolism, linking glycolysis and Krebs’ cycle and lipid metabolism [50]. In this study, PDH activity was failed to detected, whereas the decreased Pdk4 gene expression might suggest the influx of acetyl-CoA from glycolysis into the Krebs’ cycle increased in response to T2 treatment on d 5 post-weaning. Indeed, a low energy diet addition with Gln, Glu, and Asp (T2) showed lower G-6-P, F-6-P, F-1,6-P, and 3-phosphoglycerate, but higher DHAP in jejunal mucosa on d 5 post-weaning, whereas the T2 did not affect these intermediate metabolites on d 21 post-weaning. Nevertheless, a normal energy diet addition with Gln, Glu, and Asp had no effect on the levels of glycolytic metabolites on neither d 5 nor d 21 post-weaning, except for the increased F-1,6-P levels on d 5 post-weaning. Our results suggested that dietary supplementation with Gln, Glu, and Asp in either a normal energy diet or a low energy diet can replenish the Krebs’ cycle in enterocyte by converting to the intermediate metabolites. However, a low energy diet supplementation with Glu, Gln, and Asp seemed to promote glycolytic influx on d 5 instead of d 21 post-weaning. Unlike the low diet treatment, a normal diet addition with Gln, Glu, and Asp did not significantly affect the glycolysis during the weaning-period. Fatty acids oxidation also plays critical role in energy homeostasis [51]. UCP2 is involved in the regulation of mitochondrial substrate oxidation, which could be stimulated by fatty acid and glutamine [52, 53]. Several studies suggested that the role of UCP2 would be to promote oxidation of glutamine and fatty acids rather than that of the pyruvate derived from glucose [54, 55]. Indeed, a normal energy diet supplementation with Gln, Glu, and Asp raised the Ucp2 mRNA abundance but decreased Cpt1α in ileal mucosa. Although a low energy diet supplementation with Gln, Glu, and Asp did not affect the Ucp2 gene expression, it upregulated the jejunal mucosal Acox1 and ileal mucosal Cpt1α mRNA abundances. CPT1 and Acox1 play key roles in the fatty acid beta-oxidation in either mitochondria or peroxisomes, respectively. CPT1 is the key enzyme in the carnitine-dependent transport across the mitochondrial inner membrane [56], while Acox1 is the first enzyme of the fatty acid beta-oxidation in peroxisomes, which catalyzes the desaturation of acyl-CoAs to 2-trans-enoyl-CoAs [57]. Based on these results, it may be suggested that a low energy diet addition with Gln, Glu, and Asp activated the fatty acids beta-oxidation. Besides, supplementation with Gln, Glu, and Asp in a low energy diet increased jejunal mucosal NADPH, GMP and AMP levels, as well as the Pck1 gene expression on d 5 post-weaning. NADPH is mainly produced through pentose cycle, which is an alternative way of glucose use [58]. Because the current study did not detect other enzymes or intermediate metabolites involved in the pentose cycle, it is not clear whether a low energy diet supplementation with Gln, Glu, and Asp regulates the NADPH levels via the pentose cycle. Moreover, the decreased Pck1 gene expression, which is a main control point of the regulation of gluconeogenesis [59], may implicate the reduced gluconeogenesis in the T2 group on d 5 post-weaning. The higher AMP and GMP levels activates the AMPK signaling pathway, which is a key sensor of energy balance responding to low energy status and upregulating catabolic pathways and downregulating anabolic pathways [60]. Thus, it is reasonable to speculate that a low diet addition with Gln, Glu, and Asp activates the AMPK signaling pathway. A low energy diet supplementation with Gln, Glu, and Asp (T2) up-regulated the AMPK and ACC phosphorylation, as well as the SirT1 and PGC1α protein abundance in the small intestine on d 5 post-weaning, whereas it down-regulated the AMPK pathway phosphorylation levels on d 21 post-weaning. Under conditions of low energy, activated AMPK phosphorylates downstream substrates, increasing ATP levels by promoting glucose and lipids breakdown and inhibiting their synthesis and storage [7, 61]. This is consistent with the observation from energy metabolites and these enzymes’ expression mentioned above. AMPK could acutely regulate glycolysis and reduce gluconeogenesis, and controls overall lipids metabolism through direct phosphorylation of ACC and simultaneously promoting fatty acid oxidation by relieving the suppression of CPT1 [7]. Another interesting finding is that the PGC1 α gene and protein abundances were enhanced by a low energy diet supplementation with Gln, Glu, and Asp (T2). PGC1 α provides a direct link between external physiological stimuli and the regulation of mitochondrial biofunction [62]. PGC1 α regulates mitochondrial biogenesis via regulation of Nrf1 [63], which in turn activates TFAM that involved in the mDNA replication [6]. In the current study, the increased Nrf1 and Tfam gene expression in T2 treatment suggested that a low energy diet addition with Gln, Glu, and Asp stimulated the mitochondrial biogenesis in response to the acute energy crises. Unlike the low energy diet treatment, supplementation with the amino acids in a normal diet did not affect the AMPK system on d 5 post-weaning while down-regulated the phosphorylated AMPK pathway levels on d 21-post weaning. Previous studies have demonstrated that Gln could regulate enterocyte growth under normal energy conditions through activating mTOR pathway but inhibiting AMPK pathway [64, 65]. Days 2 to 5 after weaning is the most severe stage during the weaning-period [22]. Piglets on d 2 to 5 post-weaning requires more energy to rebuild the impaired intestinal mucosal barrier [66]. It is possible that Gln, Glu, and Asp could restore the small intestinal energy homeostasis via replenishing Krebs’ cycle under a normal energy diet condition, whereas it cannot provide sufficient energy sources to piglets fed with a low energy diet so that the AMPK pathway and mitochondrial biogenesis are activated by low energy conditions. While on d 21 post-weaning, the remodeling of the intestinal mucosa has been achieved, and the digestive system has adapted to utilize solid feed components to absorb and transport more energy sources [67, 68], Gln, Glu, and Asp could balance the intestinal energy homeostasis in piglets fed with a low energy diet. Conclusions The present result shows that supplementation of Gln, Glu, and Asp in a normal energy diet can promote the growth performance of weaning-piglet on d 21 post-weaning. Supplementation with Gln, Glu, and Asp in either a normal energy diet or a low energy diet improved the small intestinal mucosal barrier structure by raising jejunal villus height and goblet cells number or reducing ileal crypt depth on d 5 or 21 post-weaning. In general, Gln, Glu, and Asp could restore the small intestinal energy homeostasis via replenishing the Krebs’ cycle and down-regulate the AMPK pathway. However, when piglets under specific conditions, such as energy crisis and severe weaning-stress challenge, Gln, Glu, and Asp are not sufficient to maintain the intestinal energy balance so that the AMPK, fatty acids beta-oxidation, and mitochondrial biogenesis pathways are activated to meet the high energy demand of enterocytes of piglets. Our results aid in providing new information about the nutritional intervention of Gln, Glu and Asp for the insufficient energy intake in weaning-piglet. Abbreviations Gln: glutamine; Glu: glutamate; Asp: aspartate; AMPK: AMP-activated protein kinase; GI: gastrointestinal tract; ATP: adenosine triphosphate; LPS: lipopolysaccharide; BW: body weight; LC-MS: liquid chromatography - mass spectrometry; QC: quality control; Pdk4: pyruvate dehydrogenase kinase; Pck1: phosphoenolpyruvate carboxykinase 1; Sdh1: succinate dehydrogenase 1; Ucp2: mitochondrial uncoupling protein 2; Pparα1: peroxisome proliferator-activated receptor alpha 1; Acox1: peroxisomal acyl-coenzyme A oxidase 1; Cpt1b: carnitine palmitoyltransferase 1A; Acc: acetyl-CoA carboxylase; Sirt1: Sirtuin 1; Pgc1α: peroxisome proliferator-activated receptor gamma coactivator 1-α; Tfam: mitochondrial transcription factor A; Nrf1: nuclear respiratory factor 1; qRT-PCR: quantitative reverse transcriptase PCR; ADG: average daily gain; ADFI: average daily feed intake; DHAP: dihydroxyacetone phosphate; GMP: guanosine 5 monophosphate; F-6-P: fructose 6 phosphate; G-6-P: glucose-6-phosphate; FMN: flavin mononucleotide; c-AMP: cyclic- adenosine 5 monophosphate; NADPH: nicotinamide adenine dinucleotide phosphate hydrogen; NAD + : nicotinamide adenine dinucleotide; NADP + : nicotinamide adenine dinucleotide phosphate; ADP: adenosine 5 diphosphate; TPP: thiamine pyrophosphate; PCA: principal component analysis. Declarations Availability of data and materials The datasets generated or analyzed during this study are presented in this manuscript and available to readers. Ethics approval All animals used in this study were humanely managed according to the Chinese Guidelines for Animal Welfare. The experimental protocol was approved by the Animal Care and Use Committee of the Chinese Academy of Sciences (Beijing, China). Conflicts of interest There are no conflicts of interest to declare. Consent for publication Not applicable. Author contributions Bie Tan: Conceptualization, Methodology, Supervision. Yulong Yin: Conceptualization. Jing Wang: Investigation, Data curation, Formal analysis, Writing-Original draft preparation. Nan Wang: Investigation, Data curation. Ming Qi: Investigation, Data curation. Jianjun Li: Investigation, Data curation, Software. Funding This project was supported by the National Natural Science Foundation of China (No. 31672433, 31330075, 31501964, 31560640), Key Programs of frontier scientific research of the Chinese Academy of Sciences (QYZDY-SSW-SMC008), the Earmarked Fund for China Agriculture Research System (CARS-35), Youth Innovation Team Project of ISA, CAS(2017QNCXTD_TBE) and Postdoctoral Research Foundation of China (BX20180096). Acknowledgments We thank Changsha Lvye Biotechnology Limited Company Academician Expert Workstation, Guangdong Wangda Group Academician Workstation for Clean Feed Technology Research and Development in Swine for providing technical assistance. References [1] Pluske JR, Hampson DJ, Williams IH. Factors influencing the structure and function of the small intestine in the weaned pig: A review. Livestock production science, 1997, 51: 215-236 [2] Xiao K, Song ZH, Jiao LF , et al. 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Ingredient composition of diets 1 Item Control diet T 1 diet T 2 diet Ingredient (%) Corn 23.93 24.00 24.40 Extruded corn 35.00 35.00 35.00 Soybean 8.00 8.00 11.80 Fermented soybean 9.00 9.00 4.00 Extruded soybean 0.00 0.00 2.80 Whey powder 6.00 6.00 6.00 Fish meal 4.00 4.00 4.00 Plasma protein powder 2.00 2.00 2.00 Soybean oil 1.00 1.00 0.00 Glucose 3.00 3.00 0.00 Sucrose 2.00 2.00 0.00 L-lysine (98 %) 0.40 0.40 0.40 DL- methionine 0.11 0.11 0.11 L- threonine 0.12 0.12 0.12 L-alanine 1.59 0.00 0.00 L-glutamine 0.00 1.00 1.00 L-glutamate 0.00 0.50 0.50 L-aspartate 0.00 0.10 0.10 Carrier 0.90 0.82 0.82 Organic acid calcium 0.60 0.60 0.60 CaHPO 4 1.00 1.00 1.00 Choline chloride (50 %) 0.01 0.01 0.01 Antioxidant 0.05 0.05 0.05 Mineral premix 2 0.15 0.15 0.15 Vitamin premix 3 0.04 0.04 0.04 ZnO 0.40 0.40 0.40 Acidifier 0.70 0.70 0.70 Total 100 100 100 Nutrient composition * Digestible energy (K cal / Kg) 3445.60 3444.56 3227.00 Crude protein 19.57 19.53 19.53 Calcium 0.47 0.47 0.47 Total phosphorus 0.40 0.40 0.40 Total lysine 1.14 1.14 1.11 1 T 1 = Basal diet containing 1 % L-glutamine, 0.5 % L-glutamate, and 0.1 % L-aspartate; T 2 = energy deficiency diet containing 1 % L-glutamine, 0.5 % L-glutamate, and 0.1 % L-aspartate; the premix did not contain additional copper, zinc, antibiotics, or probiotics. 2 Mineral premix provided for 1 kg completed diet: Zn (ZnO), 50 mg; Cu (CuSO4), 20 mg; Mn (MnO), 55 mg; Fe (FeSO4), 100 mg; I (KI), 1 mg; Co (CoSO4), 2 mg; Se (Na2SeO3), 0.3 mg. 3 Vitamin premix provided for 1 kg completed diet: vitamin A, 8,255 IU; vitamin D3, 2,000 IU; vitamin E, 40 IU; vitamin B1, 2 mg; vitamin B2, 4 mg; pantothenic acid, 15 mg; vitamin B6, 10 mg; vitamin B12, 0.05 mg; nicotinic acid, 30 mg; folic acid, 2 mg; vitamin K3, 1.5 mg; biotin, 0.2 mg; choline chloride, 800 mg; and vitamin C, 100 mg. * Calculated. Table 2. Growth performance of weaning piglets 1,2 . Items Treatments SEM P -value Control T 1 T 2 Days 5 post-weaning Initial weight (kg) 7.09 7.05 7.09 0.070 0.959 BW on w5d (kg) 6.89 6.91 6.94 0.063 0.952 ADG (g/pig per day) -51.29 -34.17 -38.83 9.767 0.782 ADFI (g/pig per day) 69.14 89.20 93.87 5.140 0.127 F/G -1.35 -2.61 -2.42 0.893 0.240 Digestible energy intake (K cal/pig) 238.22 307.27 302.91 16.61 0.190 Days 21 post-weaning BW on w21d(kg) 11.58 b 13.17 a 12.07 ab 0.278 0.047 ADG (g/pig per day) 224.57 b 306.00 a 248.88 ab 13.801 0.035 ADFI (g/pig per day) 477.42 526.72 524.54 25.139 0.691 F/G 2.13 1.72 2.11 0.136 0.405 Digestible energy intake (K cal/pig) 1644.99 1814.33 1692.69 83.900 0.709 1 Control = basal diets containing 1.59 % L-alanine (iso-nitrogen); T 1 = basal diets containing 1 % glutamine, 0.5 % glutamate, 0.1 % aspartate; T 2 = energy deficiency diets containing 1 % glutamine, 0.5 % glutamate, 0.1 % aspartate; w 5 d = days 5 post-weaning at d 21 of age, w 21 d = days 21 post-weaning at d 21 of age; BW = body weight; ADG = average daily gain; ADFI = average daily feed intake; F / G = gain: feed ratio. 2 a-c Values with different letters within the same row are different ( P < 0.05); n = 6. Table 3. Jejunal and ileal morphology of weanling piglets 1, 2, 3 . Items Diet treatments Days post-weaning SEM P -value Control T 1 T 2 5 21 Diet Day Diet Day Diet × Day Jejunum Villus height (μm) 375.31 393.77 392.16 337.64 436.52 10.44 8.52 0.394 <0.001 0.049 Crypt depth (μm) 119.98 117.11 118.58 112.95 124.16 5.15 4.20 0.925 0.069 0.284 VCR 3.15 3.43 3.36 3.02 3.61 0.17 0.14 0.483 0.006 0.815 Goblet cells 10.68 ab 10.23 b 11.17 a 10.42 10.97 0.25 0.20 0.044 0.069 0.503 Lymphocyte cells 38.98 38.7 39.62 39.73 38.47 0.58 0.47 0.528 0.069 0.626 Ileum Villus height (μm) 358.17 390.65 347.49 322.49 408.39 10.21 8.34 0.015 <0.001 0.020 Crypt depth (μm) 118.32 117.98 106.56 101.34 127.23 3.48 2.84 0.037 <0.001 0.046 VCR 3.05 3.37 3.26 3.22 3.23 0.13 0.10 0.203 0.936 0.634 Goblet cells 10.53 10.82 10.72 10.53 10.84 0.32 0.26 0.816 0.403 0.455 Lymphocyte cells 40.12 40.67 39.65 40.03 40.26 0.38 0.31 0.189 0.619 0.483 1 Control = basal diets containing 1.59 % L-alanine (iso-nitrogen); T 1 = basal diets containing 1 % glutamine, 0.5 % glutamate, 0.1 % aspartate; T 2 = energy deficiency diets containing 1 % glutamine, 0.5 % glutamate, 0.1 % aspartate; w 5 d = days 5 post-weaning at d 21 of age, w 21 d = days 21 post-weaning at d 21 of age; VCR = the ratio of villus height to crypt depth. 2 The unit of goblet cells and lymphocyte is cell number per area. 3 The results were presented as main effects, and the differences among treatments or days were evaluated using Tukey HSD test. a-b Values with different letters within the same row are different ( P < 0.05); n = 6. Table S1. Primers used for real-time quantitative reverse transcription-PCR 1 . Gene Accession No. Primers β -Actin XM_003124280.3 F: 5’-GGATGCAGAAGGAGATCACG-3’ R: 5’-ATCTGCTGGAAGGTGGACAG-3’ Pdk4 NM_001159306.1 F: 5’-CTACAGCACCAACACCTGTGA-3’ R: 5’-CATCCGTTCCATATCCTGGCA-3’ Pck1 XM_005673043.3 F: 5’-TCTGGTGTACGAGGCTCTCA-3’ R: 5’-TTGCCGAAGTTGTAGCCGAA-3’ Sdh1 NM_001179714.1 F: 5’-CCGCTGGTCTCTTCCTATGG-3’ R: 5’-TGGCCCTAAACACAGGGACT-3’ Ucp2 NM_214289.1 F: 5’-CTCACCCAATGTCGCTCGTA-3’ R: 5’-ATCTCGTCTTGACCACGTCC-3’ Ppar α 1 NM_001044526.1 F: 5’-GCAGATCTCAAGTCTCTCGCC-3’ R: 5’-CCTTGTTCTGGATGCCGTTG-3’ Acox1 NM_001101028.1 F: 5’-TGTCAACTTCACCCCAACCTG -3’ R: 5’-AGGTAGGACACCATGCCACA -3’ Cpt1 α NM_001129805.1 F: 5’-GCATTTGTCCCATCTTTCGT -3’ R: 5’-GCACTGGTCCTTCTGGGATA -3’ Ampk XM_021076522.1 F: 5’-CTGGATTATGAGTGGAAGGT-3’ R: 5’-AAGTCAACAGGAGAAGAGTC-3’ Acc NM_001114269.1 F: 5’-GCCGAAACATCTCTGGGATA -3’ R: 5’-CTCCAGGACAGCACAGATCA -3’ Sirt1 NM_001145750.2 F: 5’-AACATGAAGAGGTGTGGGTG -3’ R: 5’-GAACAGGTTGCGGGAATC -3’ Pgc1 α XM_005665970.3 F: 5’-GTGAGACAGAGCCAGGTA-3’ R: 5’-TTGATGATGAAGGTGAAGGT-3’ Tfam NM_001130211.1 F: 5’-CCACCCTGAGTGGTTTTCCA-3’ R: 5’-TGCCAGTCTGCCCTATAAGC-3’ Nrf1 XM_021078993.1 F: 5’-CATGGCACTCAACAGCGAAG-3’ R: 5’-ACATGCTCACAGGGATCTGG-3’ 1 Pdk4: pyruvate dehydrogenase kinase, Pck1: phosphoenolpyruvate carboxykinase 1, Sdh1: succinate dehydrogenase 1, Ucp2: mitochondrial uncoupling protein 2, Pparα1: peroxisome proliferator-activated receptor alpha 1, Acox1: peroxisomal acyl-coenzyme A oxidase 1, Cpt1α: carnitine palmitoyltransferase 1A, Ampk, AMP-activated protein kinase, Acc: acetyl-CoA carboxylase, Sirt1: Sirtuin 1, Pgc1 α : peroxisome proliferator-activated receptor gamma coactivator 1-α, Nrf1: nuclear respiratory factor 1, Tfam: mitochondrial transcription factor A. F: forward; R: reverse. Table S2. The relative density of jejunal mucosal metabolites of weanling piglets 1, 2 . Items Diet treatments Days post-weaning SEM P -value Control T 1 T 2 5 21 Diet Day Diet Day Diet × Day AKG 1.00 1.22 1.09 1.21 0.99 0.03 0.02 <0.001 <0.001 <0.001 Oxaloacetate 1.00 1.08 1.15 0.96 1.20 0.07 0.06 0.307 0.005 0.047 Succinate 1.00 b 1.49 a 1.33 a 1.31 1.24 0.10 0.08 0.005 0.572 0.091 Phosphoenolpyruvate 1.00 a 0.55 b 0.40 b 0.68 0.62 0.11 0.09 0.001 0.575 0.299 Malate 1.00 0.99 0.97 0.97 1.01 0.02 0.01 0.353 0.053 0.367 Lactate 1.00 1.00 1.04 0.95 1.08 0.04 0.04 0.720 0.016 0.199 Aconitate 1.00 2.40 1.73 1.01 2.41 0.9 0.74 0.556 0.190 0.544 GMP 1.00 1.05 1.23 1.39 0.79 0.18 0.14 0.633 0.006 0.044 NAD + 1.00 0.85 0.82 1.02 0.76 0.1 0.08 0.388 0.028 0.279 NADH 1.00 1.53 1.97 2.00 1.00 0.45 0.37 0.326 0.062 0.100 NADPH 1.00 0.71 1.45 1.09 1.02 0.06 0.05 <0.001 0.324 <0.001 NADP + 1.00 1.03 1.03 0.88 1.15 0.12 0.10 0.981 0.059 0.295 ADP 1.00 1.27 0.94 1.09 1.05 0.11 0.09 0.080 0.770 0.970 AMP 1.00 1.09 1.54 1.46 0.96 0.11 0.09 0.003 <0.001 <0.001 Cyclic-AMP 1.00 1.03 1.01 1.05 0.98 0.05 0.04 0.897 0.159 0.546 Isocitrate 1.00 1.34 1.33 1.45 1.00 0.08 0.07 0.011 <0.001 0.011 Citrate 1.00 1.13 1.07 1.03 1.10 0.16 0.13 0.831 0.714 0.508 Pyruvate 1.00 1.20 1.17 1.24 1.01 0.06 0.05 0.071 0.003 0.046 Fumarate 1.00 1.14 1.08 1.02 1.13 0.05 0.04 0.122 0.062 0.183 ATP 1.00 1.00 1.08 1.12 0.94 0.10 0.10 0.809 0.133 0.157 GDP 1.00 0.86 0.99 0.98 0.92 0.07 0.06 0.325 0.461 0.436 GTP 1.00 0.79 0.85 0.96 0.81 0.11 0.09 0.416 0.259 0.640 TPP 1.00 0.96 0.84 0.95 0.92 0.08 0.07 0.362 0.752 0.847 FMN 1.00 1.06 1.06 1.01 1.07 0.07 0.05 0.779 0.447 0.702 F-1,6-P 1.00 3.06 1.05 2.48 0.93 0.60 0.49 0.032 0.032 0.029 F-6-P 1.00 1.02 1.07 0.85 1.21 0.10 0.08 0.870 0.004 0.017 3-phosphoglycerate 1.00 0.87 0.69 0.83 0.87 0.07 0.06 0.013 0.605 0.001 G-6-P 1.00 0.99 1.11 0.87 1.19 0.13 0.11 0.749 0.041 0.009 DHAP 1.00 0.81 1.53 1.36 0.86 0.09 0.08 <0.001 <0.001 <0.001 1 Control, basal diets containing 1.59 % L-alanine (iso-nitrogen); T1, basal diets containing 1 % glutamine, 0.5 % glutamate, 0.1 % aspartate; T2, energy deficiency diets containing 1 % glutamine, 0.5 % glutamate, 0.1 % aspartate. G-6-P: glucose-6-phosphate, F-6-P: fructose 6 phosphate, F-1,6-P: fructose 1,6 phosphate, DHAP: dihydroxyacetone phosphate, AKG: α-ketoglutarate, ATP: adenosine 5 triphosphate, ADP: adenosine 5 diphosphate, AMP: adenosine 5 monophosphate, GTP: guanosine 5 triphosphate, GDP: guanosine 5 diphosphate, GMP: guanosine 5 monophosphate, NAD + : nicotinamide adenine dinucleotide + , NADH: nicotinamide adenine dinucleotide hydrogen, NADP + : nicotinamide adenine dinucleotide phosphate, NADPH: nicotinamide adenine dinucleotide phosphate hydrogen, TPP: thiamine pyrophosphate, FMN: flavin mononucleotide. 2 The results were presented as main effects, and the differences among treatments or days were evaluated using Tukey HSD test. a-b Values with different letters within the same row are different ( P < 0.05); n = 6. Table S3. Relative expressions of gene involved in the energy metabolism 1,2 . Items Diet treatments Days post-weaning SEM P -value Control T 1 T 2 5 21 Diet Day Diet Day Diet × Day Jejunal mucosa Pdk4 0.99 1.31 0.79 0.95 1.11 0.15 0.13 0.089 0.402 0.009 Pck1 1.00 0.76 0.97 0.77 1.05 0.28 0.24 0.822 0.399 0.038 Sdh1 1.09 2.03 2.01 0.88 2.54 0.33 0.27 0.085 <0.001 0.007 Ucp2 1.00 1.02 1.92 1.54 1.08 0.54 0.43 0.393 0.466 0.407 Pparα1 0.91 1.25 1.91 1.95 0.76 0.68 0.56 0.571 0.147 0.430 Acox1 0.99 b 1.43 ab 2.82 a 2.43 1.07 0.64 0.53 0.012 0.078 0.127 Cpt1α 1.01 1.04 1.15 1.17 0.97 0.29 0.24 0.934 0.569 0.322 Ileal mucosa Pdk4 1.00 0.80 1.11 1.07 0.88 0.26 0.21 0.684 0.524 0.807 Pck1 1.01 0.62 0.20 0.67 0.54 0.26 0.21 0.098 0.659 0.748 Sdh1 1.00 0.94 1.36 0.92 1.28 0.19 0.15 0.251 0.115 0.451 Ucp2 1.00 b 3.23 a 1.68 ab 2.64 1.30 0.31 0.25 <0.001 0.001 0.052 Pparα1 0.99 1.02 1.25 1.07 1.10 0.21 0.17 0.640 0.928 0.98 Acox1 1.00 1.30 1.38 1.14 1.31 0.21 0.17 0.415 0.502 0.889 Cpt1α 1.00 b 0.81 c 1.39 a 1.13 1.01 0.05 0.04 <0.001 0.069 0.063 1 Control, basal diets containing 1.59 % L-alanine (iso-nitrogen); T1, basal diets containing 1 % glutamine, 0.5 % glutamate, 0.1 % aspartate; T2, energy deficiency diets containing 1 % glutamine, 0.5 % glutamate, 0.1 % aspartate. Pdk4: pyruvate dehydrogenase kinase, Pck1: phosphoenolpyruvate carboxykinase 1, Sdh1: succinate dehydrogenase 1, Ucp2: mitochondrial uncoupling protein 2, Pparα1: peroxisome proliferator-activated receptor alpha 1, Acox1: peroxisomal acyl-coenzyme A oxidase 1, Cpt1α: carnitine palmitoyltransferase 1A, ACC: acetyl-CoA carboxylase, SirT1: Sirtuin 1, PGC1 α : peroxisome proliferator-activated receptor gamma coactivator 1-α, Nrf1: nuclear respiratory factor 1, TFAM: mitochondrial transcription factor A. 2 The results were presented as main effects, and the differences among treatments or days were evaluated using Tukey HSD test. a-b Values with different letters within the same row are different ( P < 0.05); n = 6. Table S4. Relative gene expressions of AMPK signaling pathway of jejunal and ileal mucosa of weanling piglets 1,2 . Items Diet treatments Days post-weaning SEM P -value Control T 1 T 2 5 21 Diet Day Diet Day Diet × Day Jejunal mucosa Ampk 1.00 0.87 1.05 0.64 1.39 0.30 0.31 0.893 0.048 0.364 Acc 1.00 0.99 1.01 0.58 1.41 0.32 0.26 0.999 0.032 0.281 Sirt1 1.04 0.99 1.59 1.45 0.96 0.39 0.32 0.493 0.282 0.449 Pgc1α 0.99 a 0.37 b 0.86 a 0.55 0.89 0.14 0.11 0.013 0.038 0.105 Tfam 1.00 a 0.49 b 0.78 a 0.69 0.83 0.09 0.08 0.003 0.195 0.327 Nrf1 1.00 a 0.52 b 0.70 a 0.65 0.84 0.07 0.05 <0.001 0.016 0.189 Ileal mucosa Ampk 1.00 0.96 1.09 0.87 1.40 0.21 0.15 0.295 0.028 0.194 Acc 1.00 0.93 1.18 0.76 1.32 0.15 0.12 0.476 0.003 0.088 Sirt1 1.00 a 0.64 b 0.86 a 0.95 0.72 0.06 0.05 0.001 0.003 0.081 Pgc1α 1.00 a 0.33 b 0.74 a 0.85 0.53 0.13 0.11 0.004 0.042 0.067 Tfam 1.00 ab 0.86 b 1.22 a 1.04 1.01 0.08 0.07 0.017 0.745 0.854 Nrf1 0.99 1.02 1.12 1.06 1.03 0.09 0.07 0.548 0.763 0.617 1 Control, basal diets containing 1.59% L-alanine (iso-nitrogen); T1, basal diets containing 1% glutamine, 0.5% glutamate, 0.1% aspartate; T2, energy deficiency diets containing 1% glutamine, 0.5% glutamate, 0.1% aspartate. AMPK: AMP-activated protein kinase, ACC: acetyl-CoA carboxylase, SirT1: Sirtuin 1, PGC1 α : peroxisome proliferator-activated receptor gamma coactivator 1-α, Nrf1: nuclear respiratory factor 1, TFAM: mitochondrial transcription factor A. 2 The results were presented as main effects, and the differences among treatments or days were evaluated using Tukey HSD test. a-b Values with different letters within the same row are different ( P < 0.05); n = 6. Table S5. Relative protein abundances of AMPK signaling pathway of jejunal and ileal mucosa of weanling piglets 1,2 . Items Diet treatments Days post-weaning SEM P -value Control T 1 T 2 5 21 Diet Day Diet Day Diet × Day Jejunal mucosa AMPK 0.99 1.09 1.15 1.31 0.85 0.05 0.04 0.084 <0.001 <0.001 P-AMPK 0.99 0.67 0.87 0.95 0.74 0.05 0.04 0.001 0.002 <0.001 ACC 0.99 0.67 0.86 1.11 0.57 0.05 0.04 0.001 <0.001 <0.001 P-ACC 0.99 0.80 1.25 1.38 0.65 0.09 0.07 0.013 <0.001 0.001 SirT1 0.99 0.67 1.08 1.21 0.63 0.05 0.04 <0.001 <0.001 <0.001 PGC1a 0.99 0.88 1.12 1.36 0.64 0.09 0.07 0.172 <0.001 <0.001 Ileal mucosa AMPK 0.99 0.81 0.55 0.99 0.58 0.05 0.04 <0.001 <0.001 <0.001 P-AMPK 0.99 0.48 0.90 0.88 0.70 0.06 0.05 <0.001 0.030 0.040 ACC 0.99 b 1.34 a 1.19 ab 1.25 1.09 0.05 0.04 0.002 0.023 0.222 P-ACC 0.99 0.64 0.72 1.01 0.56 0.07 0.05 0.007 <0.001 0.004 SirT1 0.99 0.52 1.05 1.08 0.63 0.05 0.04 <0.001 <0.001 <0.001 PGC1a 0.99 0.45 1.04 1.07 0.59 0.05 0.04 <0.001 <0.001 <0.001 1 Control, basal diets containing 1.59% L-alanine (iso-nitrogen); T1, basal diets containing 1% glutamine, 0.5% glutamate, 0.1% aspartate; T2, energy deficiency diets containing 1% glutamine, 0.5% glutamate, 0.1% aspartate. AMPK: AMP-activated protein kinase, ACC: acetyl-CoA carboxylase, SirT1: Sirtuin 1, PGC1 α : peroxisome proliferator-activated receptor gamma coactivator 1-α, Nrf1: nuclear respiratory factor 1, TFAM: mitochondrial transcription factor A. 2 The results were presented as main effects, and the differences among treatments or days were evaluated using Tukey HSD test. a-b Values with different letters within the same row are different ( P < 0.05); n = 6. Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-150010","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research","associatedPublications":[],"authors":[{"id":8454162,"identity":"441064d1-3016-45b0-ae0a-a7064eb48477","order_by":0,"name":"Jing Wang","email":"","orcid":"https://orcid.org/0000-0001-8299-0359","institution":"Hunan Normal University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Jing","middleName":"","lastName":"Wang","suffix":""},{"id":8454163,"identity":"18ce542c-8cc3-4ba0-8d64-a2c02ac6a1a5","order_by":1,"name":"Nan Wang","email":"","orcid":"","institution":"Hunan Agricultural University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Nan","middleName":"","lastName":"Wang","suffix":""},{"id":8454164,"identity":"a9d9d04b-8e89-4aa1-bfe3-25338fb6030d","order_by":2,"name":"Ming Qi","email":"","orcid":"","institution":"Institute of Subtropical Agriculture Chinese Academy of Sciences","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Ming","middleName":"","lastName":"Qi","suffix":""},{"id":8454165,"identity":"46b3c20f-bd15-444c-bc76-a902d0a4800a","order_by":3,"name":"Jianjun Li","email":"","orcid":"","institution":"Institute of Subtropical Agriculture Chinese Academy of Sciences","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Jianjun","middleName":"","lastName":"Li","suffix":""},{"id":8454166,"identity":"f89daaa2-c47e-478e-b323-23ecdc5ff417","order_by":4,"name":"Bie Tan","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA0klEQVRIiWNgGAWjYBACA2YwZZMA5TMTpYWxgYEhjRQtDGAth0nQYs7OfPzBzx3n8wxuNz+TYKiwTmxgP3sArxbLZrbExt4zt4sN7hwzk2A4k57YwJOXgFeLwWEewwbettuJG24kmEkwth1ObJDgMSCghf9j49+2c0At6d8kGP8RpYWHsZm37QBQSw7QlgYitAD9Yjhbti05ceaNnGKLhGPpxm08Ofi1mPMffvDxbZtdYt+N9I03PtRYy/azn8GvBRUkADEbCepHwSgYBaNgFOAAAAVcR7NVAfENAAAAAElFTkSuQmCC","orcid":"https://orcid.org/0000-0003-0138-1530","institution":"Hunan Agricultural University","correspondingAuthor":true,"submittingAuthor":false,"prefix":"","firstName":"Bie","middleName":"","lastName":"Tan","suffix":""},{"id":8454167,"identity":"77a2204b-0d4b-494c-a143-096c397bba26","order_by":5,"name":"Yulong Yin","email":"","orcid":"","institution":"Hunan Agricultural University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Yulong","middleName":"","lastName":"Yin","suffix":""}],"badges":[],"createdAt":"2021-01-18 13:31:31","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-150010/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-150010/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":5132467,"identity":"13f201a0-89f0-4e0f-8ce4-d345de38c0fd","added_by":"auto","created_at":"2021-01-20 18:40:19","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":295300,"visible":true,"origin":"","legend":"Small intestinal morphology of weaning piglets. The representative images of scanning electron microscopy in the jejunum (A) and ileum (B) of piglets on days 5 (w 5 d), 21 (w 21 d) post-weaning at 21 d of age (magnification × 150, scale bar = 300 μm) (n = 6). (C) Histogram of the jejunal and ileal villus height and crypt depth detected by hematoxylin-eosin staining. Data are presented as the simple main effect analysis and expressed as means ± SEM, n=6. a-b Values with different lowercase letters are different (P \u003c 0.05). Control: basal diets containing 1.59 % L-alanine (iso-nitrogen); T 1: basal diet containing 1 % L-glutamine, 0.5 % L-glutamate, and 0.1 % L-aspartate; T 2: energy deficiency diet containing 1 % L-glutamine, 0.5 % L-glutamate, and 0.1 % L-aspartate.","description":"","filename":"Fig1.png","url":"https://assets-eu.researchsquare.com/files/rs-150010/v1/c306a3f33e2e37f876c115c2.png"},{"id":5132268,"identity":"d8c8c64d-bb1f-4a81-a74e-89ef20c1951a","added_by":"auto","created_at":"2021-01-20 18:34:19","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":345172,"visible":true,"origin":"","legend":"Regulation of jejunal mucosal energy metabolism by glutamine, glutamate, and aspartate in weaning piglets. (A) Heat map of the metabolites of jejunal mucosa in weaning piglets on days 5 (w 5 d), 21 (w 21 d) post-weaning. (B) Principal components analysis of jejunal mucosal metabolites in weaning piglets on days 5 (w 5 d), 21 (w 21 d) post-weaning. (C) Representative variation of energy metabolites of jejunal mucosa in weaning piglets on days 5 (w 5 d), 21 (w 21 d) post-weaning. (D) Relative mRNA expressions of pyruvate dehydrogenase kinase (Pdk4), phosphoenolpyruvate carboxykinase 1 (Pck1), and succinate dehydrogenase 1 (Sdh1) in jejunal mucosa of weaning piglets on days 5 (w 5 d), 21 (w 21 d) post-weaning. Data are presented as the simple main effect analysis and expressed as means ± SEM, n=6. a-c Values with different lowercase letters are different (P \u003c 0.05). Control: basal diets containing 1.59 % L-alanine (iso-nitrogen); T 1: basal diet containing 1 % L-glutamine, 0.5 % L-glutamate, and 0.1 % L-aspartate; T 2: energy deficiency diet containing 1 % L-glutamine, 0.5 % L-glutamate, and 0.1 % L-aspartate.","description":"","filename":"Figure2energymetabolites.png","url":"https://assets-eu.researchsquare.com/files/rs-150010/v1/e2f8f6fefbe7947ddd015c54.png"},{"id":5132397,"identity":"bdee6a34-e970-4392-abea-f1918c904bcb","added_by":"auto","created_at":"2021-01-20 18:37:19","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":403021,"visible":true,"origin":"","legend":"Relative protein abundances of AMP-activated protein kinase (AMPK) signaling pathway in jejunal (A) and ileal (B) mucosa of weaning piglets on days 5 (w 5 d), 21 (w 21 d) post-weaning. Data are presented as the simple main effect analysis and expressed as means ± SEM, n=6. a-c Values with different lowercase letters are different (P \u003c 0.05). Control: basal diets containing 1.59 % L-alanine (iso-nitrogen); T 1: basal diet containing 1 % L-glutamine, 0.5 % L-glutamate, and 0.1 % L-aspartate; T 2: energy deficiency diet containing 1 % L-glutamine, 0.5 % L-glutamate, and 0.1 % L-aspartate. ACC: acetyl-CoA carboxylase, SirT1: Sirtuin 1, PGC1α: peroxisome proliferator-activated receptor gamma coactivator 1-α.","description":"","filename":"Figure3AMPK.png","url":"https://assets-eu.researchsquare.com/files/rs-150010/v1/5e42957ec4385a3f0ed330a8.png"},{"id":5132399,"identity":"945524dc-6423-4c37-a8d2-f3fb7443b649","added_by":"auto","created_at":"2021-01-20 18:37:19","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":440205,"visible":true,"origin":"","legend":"Schematic representation of main changes of the energy metabolites and enzymes involved in the glycolysis, Krebs’ cycle, and AMP-activated protein kinase (AMPK) (dashed rectangular) pathways identified in jejunal mucosa of weaning piglets. The filled rectangular and italics represent the metabolite and enzyme are measured in current study. The solid and dashed arrows (↑, increase; ↓, decrease) in green color represent the significant changes in the T1 treatment on d 5 and 21 post-weaning, respectively. The solid and dashed arrows (↑, increase; ↓, decrease) in red color represent the significant changes in the T2 treatment on d 5 and 21 post-weaning, respectively. The solid arrows (↑, increase; ↓, decrease) in blue color represent the main effect of three treatments (Control, T1, T2) when there is no significant interaction between treatment and days. Control: basal diets containing 1.59 % L-alanine (iso-nitrogen); T 1: basal diet containing 1 % L-glutamine, 0.5 % L-glutamate, and 0.1 % L-aspartate; T 2: energy deficiency diet containing 1 % L-glutamine, 0.5 % L-glutamate, and 0.1 % L-aspartate. G-6-P: glucose-6-phosphate, F-6-P: fructose 6 phosphate, F-1,6-P: fructose 1,6 phosphate, DHAP: dihydroxyacetone phosphate, AKG: α-ketoglutarate, AMP: adenosine 5 monophosphate, GMP: guanosine 5 monophosphate, PDH: pyruvate dehydrogenase, PDK4: pyruvate dehydrogenase kinase, SDH: succinate dehydrogenase 1, PEPCK: phosphoenolpyruvate carboxykinase, CPTI: carnitine palmitoyltransferase 1, CPTII: carnitine palmitoyltransferase 2, ACC: acetyl-CoA carboxylase, SirT1: Sirtuin 1, PGC1α: peroxisome proliferator-activated receptor gamma coactivator 1-α, Nrf1: nuclear respiratory factor 1, TFAM: mitochondrial transcription factor A.","description":"","filename":"Figure4SchematicrepresentationAMPK.png","url":"https://assets-eu.researchsquare.com/files/rs-150010/v1/0cdf06f54c4df2bbc80881a7.png"},{"id":13650021,"identity":"59765184-b6de-4e82-9c51-3992d014c4b3","added_by":"auto","created_at":"2021-09-17 09:39:19","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1812512,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-150010/v1/3622f670-8c76-4989-99f5-9389a83cb5a2.pdf"}],"financialInterests":"","formattedTitle":"Glutamine, Glutamate, and Aspartate Improves Morphology and Energy Production of Small Intestine in Piglets with Different Energy Levels Diets","fulltext":[{"header":"Background","content":"\u003cp\u003eAt weaning, young pigs are subjected to removal from sow and littermates and are transported to a new environment, as well as their diet is abrupt changed from sow\u0026rsquo;s milk to solid diet, which cause low voluntary food intake and associated poor intestinal barrier structure and function [1, 2]. Undoubtedly, the weaning pigs are not eating enough food and their intestinal mucosa is failing to digest and absorb sufficient nutrients to cover their energy requirement for maintenance [3], which are major limitations to improve the growth of piglet after-weaning. Therefore, improving the energy intake from exogenous nutrition is rationally served as a promising approach to protect piglets from weaning stress.\u003c/p\u003e\n\u003cp\u003eThe intestine of piglet has notably high energy demand due to the repaid renewal of epithelium within few days [4, 5]. Hence, the epithelial cells of gastrointestinal (GI) tract require intense anabolic metabolism. It has been demonstrated that GI tract represents approximately 5% of body weight, whereas it is responsible for about 20% of whole body O\u003csub\u003e2\u003c/sub\u003e consumption [6]. Glucose and lipid are major sources for the supply and storage of energy in cells [7]. AMP-activated protein kinase (AMPK) is the master regulator of energy metabolism [6]. AMPK increases adenosine triphosphate (ATP) levels by promoting glucose and lipids breakdown and inhibiting their synthesis and storage [7]. Glycolysis, Krebs\u0026rsquo; cycle, and fatty acids beta-oxidation are the main catabolic processes for glucose and lipids [8], respectively. Although all animals obtain their biological energy from the cell-specific oxidation of fatty acids, glucose, and amino acids in diets [9], for gastrointestinal tract, glutamine (Gln), glutamate (Glu), and aspartate (Asp) are the main energy sources to maintain gut integrity and function [10, 11]. Meanwhile, numerous studies have reported that Gln, Glu, and Asp has profound impacts on the intestinal nutrition and health. Gln plays multiple roles in regulating intestinal protein turnover [12], gene expression [13], cell proliferation [14] and immune function [15]. Asp could improve growth performance of weanling piglets [16], and attenuate the intestinal injury induced by \u003cem\u003eEscherichia coli\u003c/em\u003e lipopolysaccharide (LPS) [17]. Nevertheless, a typical corn- and soybean meal-based diet cannot provide sufficient amounts of Asp, Glu, and Gln for protein synthesis in post-weaning pigs [18].\u003c/p\u003e\n\u003cp\u003eIn the present study, we hypothesize that Gln, Glu, and Asp could improve the small intestinal structure of weanling piglets via regulating the enterocyte energy metabolism. The effects of supplementation of Gln, Glu, and Asp on the intestinal morphology, energy metabolites, AMPK pathway phosphorylation were determined in weaning piglets on d 5 and d 21 post-weaning.\u003c/p\u003e"},{"header":"Methods","content":"\u003cp\u003eAll animals used in this study were humanely managed according to the Chinese Guidelines for Animal Welfare. The experimental protocol was approved by the Animal Care and Use Committee of the Chinese Academy of Sciences (Beijing, China).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003eAnimals and experimental design\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eA total of 198 piglets (Duroc \u0026times;Landrace \u0026times; Large Yorkshire) weaned at 21-d of age were assigned to 18 pens based on their body weight (BW). There were 11 piglets per pen and 6 pens per treatment. The treatment groups include: i) Control (Basal diet + 1.59% L-Alanine (Ala); iso-nitrogenous control); ii) T1 (Basal diet + 1% L-Gln + 0.5% L-Glu + 0.1% L-Asp); iii) T2 (Low energy diet + 1% L-Gln + 0.5% L-Glu + 0.1% L-Asp) (Table 1). The basal diets were formulated to meet the nutrient requirements recommended by the National Research Council (2012). A low energy level diet was obtained by removing the soybean oil, glucose, and sucrose. The dose was based on the growth performance of piglets in the preliminary experiment. All piglets were housed in an environmentally well controlled nursery facility with slatted plastic flooring and a mechanical ventilation system, and had free access to drink water.\u003c/p\u003e\n\u003cp\u003eOne piglet closed to the average body weight from each pen was slaughtered on day 5 (w5d) and 21 (w21d) post-weaning. After being stunned electrically, piglets were sacrificed and the jejunum and ileum was dissected and rinsed thoroughly with ice-cold physiological saline. The middle segments of the jejunum (2 cm) and ileum (2 cm) were cut and fixed in 2.5% glutaraldehyde or 4% formaldehyde for morphological and immunohistochemical analysis. Samples of the jejunal and ileal mucosa were scraped, immediately snap frozen in liquid nitrogen and stored at -80 \u0026deg;C for further analysis.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003eIntestinal morphology\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe segments of the jejunum and ileum fixed in 4 % formaldehyde were used to determine morphology using hematoxylin-eosin staining. After dehydration, embedding, sectioning, and staining, images were acquired at various magnifications with computer-assisted microscopy (Micro-metrics TM; Nikon ECLIPSE E200, Tokyo, Japan). Villous height, crypt depth, goblet cell and lymphocyte counts were measured by Image-Pro Plus software, Version 6.0 on images at 200- or 400-fold magnification in five randomly selected fields, respectively [19].\u003c/p\u003e\n\u003cp\u003eSegments of the jejunum and ileum at 150-fold magnification were also designated for analysis by scanning electron microscopy as described by German [20] and Liu et al. [21]. Briefly, tissue segments were fixed with 2.5 % glutaraldehyde for 2 h at 4 \u0026deg;C, and rinsed 3\u0026times;10 min in PBS at 4 \u0026deg;C. The tissues were then postfixed in 1 % osmium tetroxide for 12 h at 4 \u0026deg;C, and rinsed 3\u0026times;10 min in PBS at 4 \u0026deg;C. After samples were mounted onto stubs by means of quick-drying silver paint, the tissues were coated with gold-palladium and examined by a JEOL JSM-6360LV scanning electron microscope at 25 KV. The apparent characteristics of microvillus were observed and described.\u003c/p\u003e\n\u003cp\u003eAfter samples were mounted onto stubs by means of quick-drying silver paint, the tissues were coated with gold-palladium and examined by a JEOL SU8010 scanning electron microscope at 1.0 KV. The apparent characteristics of microvillus were observed and described.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003eMetabolomic profiling\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe ileal mucosal metabolomic profile was analyzed by liquid chromatography (LC)- mass spectrometry (MS)/MS. Briefly, 100 mg ileal mucosa was homogenized in 200 \u0026micro;L ddH\u003csub\u003e2\u003c/sub\u003eO, and mixed with 800 \u0026micro;L methanol/acetonitrile (1:1, v/v) and sonicated in ice water-bath for 30 min. After incubation in -20 \u0026deg;C for 1 h, the mixture was centrifuged at 14,000 \u003cem\u003eg\u003c/em\u003e for 15 min at 4 \u0026deg;C. The supernatant was dried by speedvac, resuspended in 100 \u0026micro;L methanol/ddH\u003csub\u003e2\u003c/sub\u003eO (1:1, v/v) for LC-MS/MS analysis. An aliquot from each sample was pooled to create quality control (QC) samples that were used to evaluate the internal standards and instrument performance. The ultra-high-performance LC (UPLC; Agilent 1290 Infinity LC, USA) was performed on a 2.1 x 100 mm ACQUITY UPLC BEH Amide column (internal diameter 1.7 \u0026mu;m; Waters, USA). The column was warmed to 45 \u0026deg;C before use. The mobile phase for UPLC analysis consisted of two solutions: (A) 15 mM ammonium acetate/H\u003csub\u003e2\u003c/sub\u003eO and (B) acetonitrile. The MS/MS (5500 QTRAP, AB SCIEX, USA) spectra was set as follows: source temperature 450 \u0026deg;C, ion Source Gas 1: 45, Ion Source Gas 2: 45, Curtain gas: 30. ionSapary Voltage Floating-4500 V.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003eReal-Time Quantitative Reverse Transcriptase PCR\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eExpressions of pyruvate dehydrogenase kinase (\u003cem\u003ePdk4\u003c/em\u003e), phosphoenolpyruvate carboxykinase 1 (\u003cem\u003ePck1\u003c/em\u003e), succinate dehydrogenase 1 (\u003cem\u003eSdh1\u003c/em\u003e), mitochondrial uncoupling protein 2 (\u003cem\u003eUcp2\u003c/em\u003e), peroxisome proliferator-activated receptor alpha 1 (\u003cem\u003ePpar\u003c/em\u003e\u003cem\u003e\u0026alpha;\u003c/em\u003e\u003cem\u003e1\u003c/em\u003e), peroxisomal acyl-coenzyme A oxidase 1 (\u003cem\u003eAcox1\u003c/em\u003e), carnitine palmitoyltransferase 1A (\u003cem\u003eCpt1a\u003c/em\u003e), acetyl-CoA carboxylase (\u003cem\u003eAcc\u003c/em\u003e), \u003cem\u003eAmpk\u003c/em\u003e, Sirtuin 1 (\u003cem\u003eSirt1\u003c/em\u003e), peroxisome proliferator-activated receptor gamma coactivator 1-\u0026alpha; (\u003cem\u003ePgc1\u0026alpha;\u003c/em\u003e), mitochondrial transcription factor A (\u003cem\u003eTfam\u003c/em\u003e), and nuclear respiratory factor 1 (\u003cem\u003eNrf1\u003c/em\u003e) mRNA in jejunal and ileal mucosa were determined by real-time quantitative reverse transcriptase PCR (real-time qRT-PCR) as described previously [22]. Primers were designed with Primer 5.0 (PREMIER Biosoft International, Palo Alto, CA) according to the gene sequence of the pig to produce an amplification product (Table S1). The comparative threshold cycle (Ct) value method (2\u003csup\u003e-\u0026Delta;\u0026Delta;Ct\u003c/sup\u003e) was employed to quantitate expression levels for target genes relative to those for the \u0026beta;-Actin. Data were expressed as the relative values to those of control piglets.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003eWestern blotting\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eJejunal and ileal mucosa samples were homogenized, and protein concentrations were measured using the bicinchoninic acid assay method with BSA as standard (Beyotime Institute of Biotechnology, Shanghai, China). All samples were adjusted to an equal concentration (70 \u0026micro;g protein). The supernatant fluid (containing tissue proteins) was diluted with 5 \u0026times; sodium dodecyl sulfate (SDS) sample buffer and heated in boiling water for 5 min and cooled down for Western blot analysis [23]. The following first antibodies were used for protein quantification: AMPK\u0026alpha; (1:1,000; Cell Signaling Technology, USA), phosphorylated AMPK\u0026alpha; (p-AMPK (Thr175); 1:1,000; Cell Signaling Technology, USA), ACC (1:1,000; Cell Signaling Technology, US), phosphorylated ACC (p-ACC (Ser79); 1:1,000; Cell Signaling Technology, USA), SirT1 (1:1,000; Cell Signaling Technology, USA), PGC1\u0026alpha; (1:1,000; Abcam, UK) and \u0026beta;-actin (1:1000; Cell Signaling Technology, USA) as well as secondary antibodies (horseradish peroxidase-conjugated goat anti-rabbit IgG; 1:5000; Boster Biological Technology, Wuhan, China). The images were detected by chemiluminescence (Millipore, Billerica, MA). Each Western blot was subjected to multiple exposures to ensure that the chemiluminescence signals were linear. Western blots were quantified by measuring the intensity of correctly sized bands using software (Alpha Imager 2200 Software; Alpha Innotech Corporation, San Leandro, CA, USA) and protein measurement was normalized to \u0026beta;-actin.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003eStatistical analysis\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe growth performance was performed by one-way ANOVA using SPSS software 19.0 (SPSS Inc., Chicago, IL). The differences among treatments were evaluated using Tukey\u0026rsquo;s test.\u003c/p\u003e\n\u003cp\u003eThe intestinal morphology, metabolomic profiles, gene and protein expression were analyzed using the general linear models\u0026rsquo; procedure, with diet treatments, days after weaning, and the interaction between treatment and day as the main effects. The results in the tables were presented as main effects. When no significant interaction between treatment and day was found, the differences among treatments or days were evaluated using Tukey HSD test (showed in Table 3, 4, 5, 6). When significant interaction between treatment and day was found, a simple main effect analysis would be performed and the differences among 3 treatments across 2 levels of days post-weaning were evaluated using Bonferroni test (showed in Figure 1, 2, 3). The developmental changes of the intestine were not the main outcome, so the differences between d 5 and d 21 post-weaning across 3 levels of treatments were not showed. Differences were declared as significant at \u003cem\u003eP\u003c/em\u003e \u0026lt; 0.05.\u003c/p\u003e"},{"header":"Results","content":"\u003cp\u003e\u003cstrong\u003e\u003cem\u003eGrowth performance\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eInitial body weight (BW), final BW, average daily gain (ADG), average daily feed intake (ADFI), and the ratio of average daily feed intake to average daily gain (F / G) were showed in Table 2. The initial BW was similar among treatments (\u003cem\u003eP\u003c/em\u003e = 0.959). On d 5 post-weaning, there were no differences on BW (\u003cem\u003eP\u003c/em\u003e = 0.952), ADG (\u003cem\u003eP\u003c/em\u003e = 0.782), ADFI (\u003cem\u003eP\u003c/em\u003e = 0.127) and F/G (\u003cem\u003eP\u003c/em\u003e = 0.240) among different treatments. On d 21 post-weaning, basal dietary supplementation with Gln, Glu, Asp (T1) improved the final BW and ADG of piglets in comparison to those in control group (\u003cem\u003eP\u003c/em\u003e \u0026lt; 0.05). However, the digestible energy intakes of weanling piglets neither on d 5 (\u003cem\u003eP\u003c/em\u003e = 0.190) nor d 21 (\u003cem\u003eP\u003c/em\u003e = 0.709) post-weaning were not significantly different.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003eJejunal and ileal morphology\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eIn Figure 1A and B, the scanning electron microscopy showed that the villi in the jejunum and ileum on d 5 post-weaning were thin and sparse, and then grew shorter, stouter and denser on d 21 post-weaning. In the jejunum, the villi heights among control, T1, and T2 groups were not showed significantly different on w 5 d, but there was an increase in villi density and villi-fold in T1 and T2 group on w 21 d. In the ileum, the increased villi densities were observed in control and T1 groups in comparison to T2 group on w 5 d, whereas villi in T1 and T2 group were denser and stouter than villi in control group on w 21 d.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eThe main effects of diet treatments and days after weaning on parameters from the intestinal hematoxylin-eosin staining were showed in Table 3. When significant interactions between treatments and days after weaning were obtained, differences among control, T1 and T2 on d 5 or d 21 post-weaning were showed in Figure 1C. In Table 3, the number of goblet cells in the jejunum of T2 group was higher than these in T1 group (\u003cem\u003eP\u003c/em\u003e \u0026lt; 0.05). There were interactions (\u003cem\u003eP\u003c/em\u003e \u0026lt; 0.05) between treatments and days on the jejunal villus height, ileal villus height and crypt depth, which means days after weaning influenced the treatments\u0026rsquo; efficacy. In Figure 1C, a low energy diet supplemented with Glu, Gln, and Asp increased (\u003cem\u003eP\u003c/em\u003e \u0026lt; 0.05) the villus height of jejunum on d 21 post-weaning and decreased (\u003cem\u003eP\u003c/em\u003e \u0026lt; 0.05) the crypt depth of ileum on d 5 post-weaning as compared to the control group, whereas it had lower (\u003cem\u003eP\u003c/em\u003e \u0026lt; 0.05) ileal villus height than the control and T1 groups on d 5 post-weaning. A basal diet supplemented with Gln, Glu, and Asp increased the villus height of ileum on d 21 post-weaning compared to the control group.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003eMetabolomic profiling of jejunal mucosa of weanling piglets\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe jejunal mucosa energy metabolite fingerprints were presented in Figure 2A and Figure 2B. Piglets on d 5 and d 21 postweaning could be readily differentiated by the heat map of their jejunal mucosa energy metabolites (Figure 2A). Piglets on d 5 post-weaning showed more abundances of dihydroxyacetone phosphate (DHAP), isocitrate, aconitate, citrate, guanosine 5 monophosphate (GMP), fructose 6 phosphate (F-6-P), and glucose-6-phosphate (G-6-P) in jejunal mucosa, while piglets on d 21 post-weaning showed more abundances of fumarate, malate, flavin mononucleotide (FMN), cyclic- adenosine 5 monophosphate (c-AMP), AMP, nicotinamide adenine dinucleotide phosphate hydrogen (NADPH), nicotinamide adenine dinucleotide phosphate (NADP\u003csup\u003e+\u003c/sup\u003e), nicotinamide adenine dinucleotide (NAD\u003csup\u003e+\u003c/sup\u003e), adenosine 5 diphosphate (ADP), and thiamine pyrophosphate(TPP) in jejunal mucosa. The first two components of a principal component analysis (PCA) showed that the jejunal mucosal energy metabolites of control and T1 treatments on d 5 post-weaning were comparable, while T2 treatment on d 5 post-weaning was more dispersive and different from control and T1 treatments. On d 21 post-weaning, energy metabolites in T1 and T2 treatments were comparable, which were different from these in the control group.\u003c/p\u003e\n\u003cp\u003eThe main effects of diet treatments and days after weaning on metabolites\u0026rsquo; levels of jejunal mucosa were showed in Table S2. When significant interactions between treatments and days after weaning were found, differences among control, T1 and T2 on d 5 or d 21 post-weaning were showed in Figure 2C. In Table S2, supplementation with Gln, Glu, and Asp in both basal and low energy diets increased (\u003cem\u003eP\u003c/em\u003e \u0026lt; 0.05) the succinate level but decreased (\u003cem\u003eP\u003c/em\u003e \u0026lt; 0.05) the phosphoenolpyruvate level in jejunal mucosa of weanling piglets. Days post-weaning had influences on the diet treatments\u0026rsquo; efficacy (\u003cem\u003eP \u003csub\u003einteraction\u003c/sub\u003e\u003c/em\u003e \u0026lt; 0.05). As shown in Figure 2C, a basal diet supplemented with Gln, Glu, and Asp (T1) increased (\u003cem\u003eP\u003c/em\u003e \u0026lt; 0.05) the pyruvate, \u0026alpha;-ketoglutarate (AKG), isocitrate, and fructose 1,6 phosphate (F-1,6-P) levels but decreased (\u003cem\u003eP\u003c/em\u003e \u0026lt; 0.05) the NADPH level on d 5 post-weaning compared to control group. A low energy diet supplemented with Gln, Glu, and Asp (T2) increased (\u003cem\u003eP\u003c/em\u003e \u0026lt; 0.05) the pyruvate, AKG, isocitrate and GMP levels compared to the control group and increased (\u003cem\u003eP\u003c/em\u003e \u0026lt; 0.05) DHAP, NADPH, and AMP levels as compared to control and T1 groups but reduced (\u003cem\u003eP\u003c/em\u003e \u0026lt; 0.05) 3-phosphoglycerate, F-6-P, and G-6-P levels on d 5 post-weaning. On d 21 post-weaning, T1 and T2 treatments showed higher (\u003cem\u003eP\u003c/em\u003e \u0026lt; 0.05) oxaloacetate level than the control group.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003eExpression of gene involved in energy metabolism of jejunal and ileal mucosa of piglets\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe gene expressions of several key enzymes of energy metabolism were presented in Table S3 and Figure 2D. The main effect in Table S3 showed T1 treatment increased (\u003cem\u003eP\u003c/em\u003e \u0026lt; 0.05) the ileal \u003cem\u003eUcp2\u003c/em\u003e mRNA level compared to the control group but decreased (\u003cem\u003eP\u003c/em\u003e \u0026lt; 0.05) the ileal \u003cem\u003eCpt1a\u003c/em\u003e mRNA level compared to control and T2 groups. T2 treatment increased (\u003cem\u003eP\u003c/em\u003e \u0026lt; 0.05) the jejunal \u003cem\u003eAcox1\u003c/em\u003e and ileal \u003cem\u003eCpt1a\u003c/em\u003e mRNA levels in comparison with the control group. Days post-weaning had influences on the diet treatments\u0026rsquo; efficacy (\u003cem\u003eP \u003csub\u003einteraction\u003c/sub\u003e\u003c/em\u003e \u0026lt; 0.05; Figure 2D). On d 5 post-weaning, a low energy diet supplemented with Gln, Glu, and Asp had lower (\u003cem\u003eP\u003c/em\u003e \u0026lt; 0.05) \u003cem\u003ePdk4\u003c/em\u003e and \u003cem\u003ePck1\u003c/em\u003emRNA abundances than control or T1 groups. On d 21 post-weaning, a basal diet supplemented with Gln, Glu, and Asp enhanced (\u003cem\u003eP\u003c/em\u003e \u0026lt; 0.05) the \u003cem\u003eSdh1\u003c/em\u003e mRNA abundance in comparison to these in the control group. A low energy diet supplemented with Gln, Glu, and Asp increased (\u003cem\u003eP\u003c/em\u003e \u0026lt; 0.05) the \u003cem\u003ePck1\u003c/em\u003e and \u003cem\u003eSdh1\u003c/em\u003e mRNA abundances as compared to T1 or control groups, respectively.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003emRNA and protein abundances of AMP-activated protein kinase signaling pathway in jejunal and ileal mucosa of piglets\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe relative mRNA and protein abundances of AMPK pathway in the jejunal and ileal mucosa were presented in Table S4, S5 and Figure 3, respectively. The main effects on AMPK pathway gene expressions in Table S4 showed a basal diet supplemented with Gln, Glu and Asp (T1) lowered (\u003cem\u003eP\u003c/em\u003e \u0026lt; 0.05) jejunal \u003cem\u003ePgc1\u0026alpha;\u003c/em\u003e, \u003cem\u003eTfam\u003c/em\u003e, and \u003cem\u003eNrf1\u003c/em\u003e and ileal \u003cem\u003eSirt1\u003c/em\u003e,\u003cem\u003e Pgc1\u0026alpha;\u003c/em\u003e mRNA levels compared to control and T2 group, as well as ileal \u003cem\u003eTfam\u003c/em\u003e mRNA level compared to the T2 group. The main effects on AMPK pathway protein abundances in Table S5 showed T1 treatment enhanced (\u003cem\u003eP\u003c/em\u003e \u0026lt; 0.05) ileal ACC protein abundance as compared to that in the control group. Days post-weaning did not affect the diet treatments\u0026rsquo; efficacy on the AMPK pathway gene expression (\u003cem\u003eP \u003csub\u003einteraction\u003c/sub\u003e\u003c/em\u003e \u0026gt; 0.05) but had influenced the treatments\u0026rsquo; efficacy on AMPK pathway protein abundances (\u003cem\u003eP \u003csub\u003einteraction\u003c/sub\u003e\u003c/em\u003e \u0026lt; 0.05). As shown in Figure 3, a basal diet supplemented with Gln, Glu and Asp (T1) enhanced (\u003cem\u003eP\u003c/em\u003e \u0026lt; 0.05) the jejunal AMPK\u0026alpha; protein abundance compared to the control group but reduced (\u003cem\u003eP\u003c/em\u003e \u0026lt; 0.05) the ileal p-AMPK\u0026alpha; and PGC1\u0026alpha; protein abundances as compared to control and T2 groups, respectively, on d 5 post-weaning. A low energy diet supplemented with Gln, Glu and Asp (T2) reduced (\u003cem\u003eP\u003c/em\u003e \u0026lt; 0.05) the ileal AMPK\u0026alpha; protein abundance compared to control and T1 groups, while increased (\u003cem\u003eP\u003c/em\u003e \u0026lt; 0.05) jejunal and ileal SirT1 and PGC1\u0026alpha;, as well as jejunal ACC and p-ACC protein abundances compared to control or T1 groups on d 5 post-weaning. On d 21 post-weaning, T1 and T2 treatments lowered (\u003cem\u003eP\u003c/em\u003e \u0026lt; 0.05) the jejunal ACC, ileal AMPK\u0026alpha; and PGC1\u0026alpha;, as well as jejunal and ileal p-AMPK\u0026alpha;, p-ACC, and SirT1 protein abundances as compared to the control groups. Meanwhile, T1 treatment showed lower (\u003cem\u003eP\u003c/em\u003e \u0026lt; 0.05) jejunal and ileal PGC1\u0026alpha; and ileal SirT1 protein abundances than these in the T2 group.\u003c/p\u003e"},{"header":"Discussion","content":"\u003cp\u003eThe mammalian intestine plays a key role in nutrients intake and has a high rate of energy expenditure because the digestion and absorption processes are highly dependent on energy [24, 25].Weaning results in a growth check caused by a period of low feed intake, which also reduces the energy intake of intestine of piglets [26, 27]. Successful adaptation to these changes requires profound morphological and energy metabolic adjustments of the gastrointestinal tract [28]. Gln, Glu and Asp were used to help the faster restitution of mucosal barrier after stress injury, and to improve the intestinal energy metabolism of piglets with different energy levels diets in this study.\u003c/p\u003e\n\u003cp\u003eThe most serious damages in piglets during weaning-stress occurs on d 2 to d 5 post-weaning [25, 29], and is usually associated with a dramatic reduction of feed intake resulting in marked changes in the growth pattern on body weight [28]. In the present study, supplementation with 1 % Gln, 0.5 % Glu, and 0.1 % Asp in a normal energy level diet enhanced the BW and ADG on d 21 post-weaning. The beneficial effect of Gln, Glu and Asp on improving the growth-suppression induced by weaning stress in our study was consistent with previous researches, which have shown that Glu and Asp exert positive effects on growth performance of animals in healthy and oxidative-stress conditions [30, 31] [32, 33]. Although low energy diet did not significantly increase the ADFI of weanling piglets, the digestive energy intake in T2 group was comparable with these in control and T1 groups. Piglets in low energy diet treatment increased their satiety through increasing food intake.\u003c/p\u003e\n\u003cp\u003eIn addition to poor growth performance, the growth-check induced by weaning-stress is accompanied by impaired mucosal integrity, villous atrophy, an increased rate of cell loss, or a decreased rate of crypt-cell production [34, 35]. Supplementation with Gln, Glu, and Asp in normal energy diet increased ileal villus height on d 21 post-weaning. The major reasons for villus atrophy are deficient enteral nutrition and reduced energy intake independent of diet type [36, 37]. We found that a low energy diet supplemented with Gln, Glu, and Asp could alleviate impaired intestinal structure by raising jejunal villus height and goblet cells number or reducing ileal crypt depth on either d 5 or d 21 post-weaning. The critical roles of these three amino acids in intestine have been reported in many well-designed studies, for example, Gln could increase the villus length of jejunum [38], and Asp could alleviate the intestinal barrier damage [32, 39, 40]. The results in our current study indicated that Gln, Glu, and Asp could improve the intestinal barrier structure of weanling piglets fed either a low energy diet or a normal energy diet, which led us to further explore the mechanism of these three amino acids regulating energy homeostasis in the small intestine.\u003c/p\u003e\n\u003cp\u003eATP production and utilization are very active in pig enterocytes. The relative abundances of intermediate metabolites involved in the energy metabolism of the small intestine of piglets on d 5 and d 21 post-weaning were not comparable, and the regulation of Gln, Glu, and Asp on energy metabolites varied between d 5 and d 21 post-weaning. It may indicate that the energy metabolism in the small intestine changes during the weaning period [41], and the pattern of metabolism has different responses to amino acids. It is well established that dietary Gln, Glu, and Asp plus arterial Gln provide up to 80 % of ATP to the small intestine mucosa in mammals [11, 42, 43]. After these three amino acids are absorbed into enterocytes, they could be metabolized and oxidized via the Krebs\u0026rsquo; cycle to water and CO2 yielding ATP [44]. The oxidative steps of Gln and Glu in pig enterocytes are similar. Gln firstly enter the mitochondrial in order to be degraded to Glu and ammonia by the mitochondrial glutaminase [45]. Glu could be transaminated to produce AKG, which is the key molecular of the Krebs\u0026rsquo; cycle [46]. Asp replenished the Krebs cycle by being converted to oxaloacetate [44]. In the present study, supplementation with Gln, Glu, and Asp either in a regular energy diet or a low energy diet could replenish the jejunal Krebs\u0026rsquo; cycle, manifested as increased pyruvate, oxaloacetate, isocitrate, AKG, and succinate levels on d 5 or d 21 post-weaning. Meanwhile, supplementation with Gln, Glu and Asp in either regular energy diet or low energy diet enhanced \u003cem\u003eSdh1\u003c/em\u003e mRNA abundance on d 21 post-weaning, whereas low energy diet supplemented with Gln, Glu, and Asp declined \u003cem\u003ePdk4\u003c/em\u003e mRNA abundance on d 5 post-weaning. SDH1 is part of Krebs\u0026rsquo; cycle and oxidizes succinate to fumarate [47]. The higher Sdh1 expression was consistent with the higher succinate level in the jejunal mucosa. \u003cem\u003eSdh1\u003c/em\u003e is responsible to promote the oxidation of succinate and prevents the accumulation of succinate in the mitochondria [48].PDK4 is a regulator of pyruvate dehydrogenase (PDH), as it could inactivate PDH by phosphorylation [49]. PDH represents a cornerstone in cellular energy metabolism, linking glycolysis and Krebs\u0026rsquo; cycle and lipid metabolism [50]. In this study, PDH activity was failed to detected, whereas the decreased \u003cem\u003ePdk4\u003c/em\u003e gene expression might suggest the influx of acetyl-CoA from glycolysis into the Krebs\u0026rsquo; cycle increased in response to T2 treatment on d 5 post-weaning. Indeed, a low energy diet addition with Gln, Glu, and Asp (T2) showed lower G-6-P, F-6-P, F-1,6-P, and 3-phosphoglycerate, but higher DHAP in jejunal mucosa on d 5 post-weaning, whereas the T2 did not affect these intermediate metabolites on d 21 post-weaning. Nevertheless, a normal energy diet addition with Gln, Glu, and Asp had no effect on the levels of glycolytic metabolites on neither d 5 nor d 21 post-weaning, except for the increased F-1,6-P levels on d 5 post-weaning. Our results suggested that dietary supplementation with Gln, Glu, and Asp in either a normal energy diet or a low energy diet can replenish the Krebs\u0026rsquo; cycle in enterocyte by converting to the intermediate metabolites. However, a low energy diet supplementation with Glu, Gln, and Asp seemed to promote glycolytic influx on d 5 instead of d 21 post-weaning. Unlike the low diet treatment, a normal diet addition with Gln, Glu, and Asp did not significantly affect the glycolysis during the weaning-period.\u003c/p\u003e\n\u003cp\u003eFatty acids oxidation also plays critical role in energy homeostasis [51]. UCP2 is involved in the regulation of mitochondrial substrate oxidation, which could be stimulated by fatty acid and glutamine [52, 53]. Several studies suggested that the role of UCP2 would be to promote oxidation of glutamine and fatty acids rather than that of the pyruvate derived from glucose [54, 55]. Indeed, a normal energy diet supplementation with Gln, Glu, and Asp raised the \u003cem\u003eUcp2\u003c/em\u003e mRNA abundance but decreased \u003cem\u003eCpt1\u0026alpha;\u003c/em\u003e in ileal mucosa. Although a low energy diet supplementation with Gln, Glu, and Asp did not affect the \u003cem\u003eUcp2\u003c/em\u003e gene expression, it upregulated the jejunal mucosal \u003cem\u003eAcox1\u003c/em\u003e and ileal mucosal \u003cem\u003eCpt1\u0026alpha;\u003c/em\u003e mRNA abundances. CPT1 and Acox1 play key roles in the fatty acid beta-oxidation in either mitochondria or peroxisomes, respectively. CPT1 is the key enzyme in the carnitine-dependent transport across the mitochondrial inner membrane [56], while Acox1 is the first enzyme of the fatty acid beta-oxidation in peroxisomes, which catalyzes the desaturation of acyl-CoAs to 2-trans-enoyl-CoAs [57]. Based on these results, it may be suggested that a low energy diet addition with Gln, Glu, and Asp activated the fatty acids beta-oxidation. Besides, supplementation with Gln, Glu, and Asp in a low energy diet increased jejunal mucosal NADPH, GMP and AMP levels, as well as the \u003cem\u003ePck1\u003c/em\u003e gene expression on d 5 post-weaning. NADPH is mainly produced through pentose cycle, which is an alternative way of glucose use [58]. Because the current study did not detect other enzymes or intermediate metabolites involved in the pentose cycle, it is not clear whether a low energy diet supplementation with Gln, Glu, and Asp regulates the NADPH levels via the pentose cycle. Moreover, the decreased \u003cem\u003ePck1\u003c/em\u003e gene expression, which is a main control point of the regulation of gluconeogenesis [59], may implicate the reduced gluconeogenesis in the T2 group on d 5 post-weaning. The higher AMP and GMP levels activates the AMPK signaling pathway, which is a key sensor of energy balance responding to low energy status and upregulating catabolic pathways and downregulating anabolic pathways [60]. Thus, it is reasonable to speculate that a low diet addition with Gln, Glu, and Asp activates the AMPK signaling pathway.\u003c/p\u003e\n\u003cp\u003eA low energy diet supplementation with Gln, Glu, and Asp (T2) up-regulated the AMPK and ACC phosphorylation, as well as the SirT1 and PGC1\u0026alpha; protein abundance in the small intestine on d 5 post-weaning, whereas it down-regulated the AMPK pathway phosphorylation levels on d 21 post-weaning. Under conditions of low energy, activated AMPK phosphorylates downstream substrates, increasing ATP levels by promoting glucose and lipids breakdown and inhibiting their synthesis and storage [7, 61]. This is consistent with the observation from energy metabolites and these enzymes\u0026rsquo; expression mentioned above. AMPK could acutely regulate glycolysis and reduce gluconeogenesis, and controls overall lipids metabolism through direct phosphorylation of ACC and simultaneously promoting fatty acid oxidation by relieving the suppression of CPT1 [7]. Another interesting finding is that the PGC1\u003cem\u003e\u0026alpha;\u003c/em\u003e gene and protein abundances were enhanced by a low energy diet supplementation with Gln, Glu, and Asp (T2). PGC1\u003cem\u003e\u0026alpha;\u003c/em\u003e provides a direct link between external physiological stimuli and the regulation of mitochondrial biofunction [62]. PGC1\u003cem\u003e\u0026alpha;\u003c/em\u003e regulates mitochondrial biogenesis via regulation of Nrf1 [63], which in turn activates TFAM that involved in the mDNA replication [6]. In the current study, the increased \u003cem\u003eNrf1\u003c/em\u003e and \u003cem\u003eTfam\u003c/em\u003e gene expression in T2 treatment suggested that a low energy diet addition with Gln, Glu, and Asp stimulated the mitochondrial biogenesis in response to the acute energy crises. Unlike the low energy diet treatment, supplementation with the amino acids in a normal diet did not affect the AMPK system on d 5 post-weaning while down-regulated the phosphorylated AMPK pathway levels on d 21-post weaning. Previous studies have demonstrated that Gln could regulate enterocyte growth under normal energy conditions through activating mTOR pathway but inhibiting AMPK pathway [64, 65]. Days 2 to 5 after weaning is the most severe stage during the weaning-period [22]. Piglets on d 2 to 5 post-weaning requires more energy to rebuild the impaired intestinal mucosal barrier [66]. It is possible that Gln, Glu, and Asp could restore the small intestinal energy homeostasis via replenishing Krebs\u0026rsquo; cycle under a normal energy diet condition, whereas it cannot provide sufficient energy sources to piglets fed with a low energy diet so that the AMPK pathway and mitochondrial biogenesis are activated by low energy conditions. While on d 21 post-weaning, the remodeling of the intestinal mucosa has been achieved, and the digestive system has adapted to utilize solid feed components to absorb and transport more energy sources [67, 68], Gln, Glu, and Asp could balance the intestinal energy homeostasis in piglets fed with a low energy diet.\u003c/p\u003e"},{"header":"Conclusions","content":"\u003cp\u003eThe present result shows that supplementation of Gln, Glu, and Asp in a normal energy diet can promote the growth performance of weaning-piglet on d 21 post-weaning. Supplementation with Gln, Glu, and Asp in either a normal energy diet or a low energy diet improved the small intestinal mucosal barrier structure by raising jejunal villus height and goblet cells number or reducing ileal crypt depth on d 5 or 21 post-weaning. In general, Gln, Glu, and Asp could restore the small intestinal energy homeostasis via replenishing the Krebs\u0026rsquo; cycle and down-regulate the AMPK pathway. However, when piglets under specific conditions, such as energy crisis and severe weaning-stress challenge, Gln, Glu, and Asp are not sufficient to maintain the intestinal energy balance so that the AMPK, fatty acids beta-oxidation, and mitochondrial biogenesis pathways are activated to meet the high energy demand of enterocytes of piglets. Our results aid in providing new information about the nutritional intervention of Gln, Glu and Asp for the insufficient energy intake in weaning-piglet.\u003c/p\u003e"},{"header":"Abbreviations","content":"\u003cp\u003eGln: glutamine; Glu: glutamate; Asp: aspartate; AMPK: AMP-activated protein kinase; GI: gastrointestinal tract; ATP: adenosine triphosphate; LPS: lipopolysaccharide; BW: body weight; LC-MS: liquid chromatography - mass spectrometry; QC: quality control; Pdk4: pyruvate dehydrogenase kinase; Pck1: phosphoenolpyruvate carboxykinase 1; Sdh1: succinate dehydrogenase 1; Ucp2: mitochondrial uncoupling protein 2; Ppar\u0026alpha;1: peroxisome proliferator-activated receptor alpha 1; Acox1: peroxisomal acyl-coenzyme A oxidase 1; Cpt1b: carnitine palmitoyltransferase 1A; Acc: acetyl-CoA carboxylase; Sirt1: Sirtuin 1; Pgc1\u0026alpha;: peroxisome proliferator-activated receptor gamma coactivator 1-\u0026alpha;; Tfam: mitochondrial transcription factor A; Nrf1: nuclear respiratory factor 1; qRT-PCR: quantitative reverse transcriptase PCR; ADG: average daily gain; ADFI: average daily feed intake; DHAP: dihydroxyacetone phosphate; GMP: guanosine 5 monophosphate; F-6-P: fructose 6 phosphate; G-6-P: glucose-6-phosphate; FMN: flavin mononucleotide; c-AMP: cyclic- adenosine 5 monophosphate; NADPH: nicotinamide adenine dinucleotide phosphate hydrogen; NAD\u003csup\u003e+\u003c/sup\u003e: nicotinamide adenine dinucleotide; NADP\u003csup\u003e+\u003c/sup\u003e: nicotinamide adenine dinucleotide phosphate; ADP: adenosine 5 diphosphate; TPP: thiamine pyrophosphate; PCA: principal component analysis.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAvailability of data and materials\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe datasets generated or analyzed during this study are presented in this manuscript and available to readers.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthics approval\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll animals used in this study were humanely managed according to the Chinese Guidelines for Animal Welfare. The experimental protocol was approved by the Animal Care and Use Committee of the Chinese Academy of Sciences (Beijing, China).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConflicts of interest\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThere are no conflicts of interest to declare.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent for publication\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthor contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eBie Tan:\u003c/strong\u003e Conceptualization, Methodology, Supervision. \u003cstrong\u003eYulong Yin:\u003c/strong\u003e Conceptualization. \u003cstrong\u003eJing Wang:\u003c/strong\u003e Investigation, Data curation, Formal analysis, Writing-Original draft preparation. \u003cstrong\u003eNan Wang: \u003c/strong\u003eInvestigation, Data curation. \u003cstrong\u003eMing Qi:\u003c/strong\u003e Investigation, Data curation. \u003cstrong\u003eJianjun Li:\u003c/strong\u003e Investigation, Data curation, Software.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis project was supported by the National Natural Science Foundation of China (No. 31672433, 31330075, 31501964, 31560640), Key Programs of frontier scientific research of the Chinese Academy of Sciences (QYZDY-SSW-SMC008), the Earmarked Fund for China Agriculture Research System (CARS-35), Youth Innovation Team Project of ISA, CAS(2017QNCXTD_TBE) and Postdoctoral Research Foundation of China (BX20180096).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgments\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eWe thank Changsha Lvye Biotechnology Limited Company Academician Expert Workstation, Guangdong Wangda Group Academician Workstation for Clean Feed Technology Research and Development in Swine for providing technical assistance.\u003c/p\u003e"},{"header":"References","content":"\u003cp\u003e[1] Pluske JR, Hampson DJ, Williams IH. 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Journal of animal science and biotechnology, 2013, 4: 7\u003c/p\u003e\n\u003cp\u003e[45] Wang B, Wu G, Zhou Z\u003cem\u003e, et al.\u003c/em\u003e Glutamine and intestinal barrier function. Amino acids, 2015, 47: 2143-2154\u003c/p\u003e\n\u003cp\u003e[46] Blachier F, Boutry C, Bos C\u003cem\u003e, et al.\u003c/em\u003e Metabolism and functions of l-glutamate in the epithelial cells of the small and large intestines. The American journal of clinical nutrition, 2009, 90: 814S-821S\u003c/p\u003e\n\u003cp\u003e[47] Rutter J, Winge DR, Schiffman JD. Succinate dehydrogenase\u0026ndash;assembly, regulation and role in human disease. Mitochondrion, 2010, 10: 393-401\u003c/p\u003e\n\u003cp\u003e[48] Connors J, Dawe N, Van Limbergen J. The role of succinate in the regulation of intestinal inflammation. Nutrients, 2019, 11: 25\u003c/p\u003e\n\u003cp\u003e[49] Lundsgaard A-M, Fritzen AM, Kiens B. Exercise physiology in men and women. Principles of gender-specific medicine, 2017, 525-542,\u003c/p\u003e\n\u003cp\u003e[50] Lazzarino G, Amorini AM, Signoretti S\u003cem\u003e, et al.\u003c/em\u003e Pyruvate dehydrogenase and tricarboxylic acid cycle enzymes are sensitive targets of traumatic brain injury induced metabolic derangement. International journal of molecular sciences, 2019, 20: 5774\u003c/p\u003e\n\u003cp\u003e[51] Houten SM, Wanders RJ. A general introduction to the biochemistry of mitochondrial fatty acid \u0026beta;-oxidation. Journal of inherited metabolic disease, 2010, 33: 469-477\u003c/p\u003e\n\u003cp\u003e[52] Hurtaud C, Gelly C, Chen Z\u003cem\u003e, et al.\u003c/em\u003e Glutamine stimulates translation of uncoupling protein 2mrna. Cellular and Molecular Life Sciences, 2007, 64: 1853-1860\u003c/p\u003e\n\u003cp\u003e[53] Bouillaud F, Alves-Guerra M-C, Ricquier D. Ucps, at the interface between bioenergetics and metabolism. 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Endocrinology, 2020, 161: bqz046\u003c/p\u003e\n\u003cp\u003e[57] Bouagnon AD, Lin L, Srivastava S\u003cem\u003e, et al.\u003c/em\u003e Intestinal peroxisomal fatty acid \u0026beta;-oxidation regulates neural serotonin signaling through a feedback mechanism. PLoS Biology, 2019, 17: e3000242\u003c/p\u003e\n\u003cp\u003e[58] Werner C, Doenst T, Schwarzer M. Metabolic pathways and cycles. The scientist's guide to cardiac metabolism, 2016, 39-55,\u003c/p\u003e\n\u003cp\u003e[59] Burgess SC, He T, Yan Z\u003cem\u003e, et al.\u003c/em\u003e Cytosolic phosphoenolpyruvate carboxykinase does not solely control the rate of hepatic gluconeogenesis in the intact mouse liver. Cell metabolism, 2007, 5: 313-320\u003c/p\u003e\n\u003cp\u003e[60] Carling D. Ampk hierarchy: A matter of space and time. Cell research, 2019, 29: 425-426\u003c/p\u003e\n\u003cp\u003e[61] Chen L, Wang J, You Q\u003cem\u003e, et al.\u003c/em\u003e Activating ampk to restore tight junction assembly in intestinal epithelium and to attenuate experimental colitis by metformin. Frontiers in pharmacology, 2018, 9: 761\u003c/p\u003e\n\u003cp\u003e[62] Gureev AP, Shaforostova EA, Popov VN. Regulation of mitochondrial biogenesis as a way for active longevity: Interaction between the nrf2 and pgc-1\u0026alpha; signaling pathways. Frontiers in Genetics, 2019, 10: 435\u003c/p\u003e\n\u003cp\u003e[63] Finck BN, Kelly DP. Peroxisome proliferator\u0026ndash;activated receptor \u0026gamma; coactivator-1 (pgc-1) regulatory cascade in cardiac physiology and disease. Circulation, 2007, 115: 2540-2548\u003c/p\u003e\n\u003cp\u003e[64] Yi D, Hou Y, Wang L\u003cem\u003e, et al.\u003c/em\u003e L-glutamine enhances enterocyte growth via activation of the mtor signaling pathway independently of ampk. 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Journal of animal science and biotechnology, 2016, 7: 4\u003c/p\u003e"},{"header":"Tables","content":"\u003cp\u003e\u003cstrong\u003eTable 1. Ingredient composition of diets \u003csup\u003e1\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"1\"\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd width=\"225\"\u003e\n\u003cp\u003eItem\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"113\"\u003e\n\u003cp\u003eControl diet\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"113\"\u003e\n\u003cp\u003eT 1 diet\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"102\"\u003e\n\u003cp\u003eT 2 diet\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd colspan=\"4\" width=\"553\"\u003e\n\u003cp\u003eIngredient (%)\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"225\"\u003e\n\u003cp\u003eCorn\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"113\"\u003e\n\u003cp\u003e23.93\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"113\"\u003e\n\u003cp\u003e24.00\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"102\"\u003e\n\u003cp\u003e24.40\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"225\"\u003e\n\u003cp\u003eExtruded corn\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"113\"\u003e\n\u003cp\u003e35.00\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"113\"\u003e\n\u003cp\u003e35.00\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"102\"\u003e\n\u003cp\u003e35.00\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"225\"\u003e\n\u003cp\u003eSoybean\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"113\"\u003e\n\u003cp\u003e8.00\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"113\"\u003e\n\u003cp\u003e8.00\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"102\"\u003e\n\u003cp\u003e11.80\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"225\"\u003e\n\u003cp\u003eFermented soybean\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"113\"\u003e\n\u003cp\u003e9.00\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"113\"\u003e\n\u003cp\u003e9.00\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"102\"\u003e\n\u003cp\u003e4.00\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"225\"\u003e\n\u003cp\u003eExtruded soybean\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"113\"\u003e\n\u003cp\u003e0.00\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"113\"\u003e\n\u003cp\u003e0.00\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"102\"\u003e\n\u003cp\u003e2.80\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"225\"\u003e\n\u003cp\u003eWhey powder\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"113\"\u003e\n\u003cp\u003e6.00\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"113\"\u003e\n\u003cp\u003e6.00\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"102\"\u003e\n\u003cp\u003e6.00\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"225\"\u003e\n\u003cp\u003eFish meal\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"113\"\u003e\n\u003cp\u003e4.00\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"113\"\u003e\n\u003cp\u003e4.00\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"102\"\u003e\n\u003cp\u003e4.00\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"225\"\u003e\n\u003cp\u003ePlasma protein powder\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"113\"\u003e\n\u003cp\u003e2.00\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"113\"\u003e\n\u003cp\u003e2.00\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"102\"\u003e\n\u003cp\u003e2.00\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"225\"\u003e\n\u003cp\u003eSoybean oil\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"113\"\u003e\n\u003cp\u003e1.00\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"113\"\u003e\n\u003cp\u003e1.00\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"102\"\u003e\n\u003cp\u003e0.00\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"225\"\u003e\n\u003cp\u003eGlucose\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"113\"\u003e\n\u003cp\u003e3.00\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"113\"\u003e\n\u003cp\u003e3.00\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"102\"\u003e\n\u003cp\u003e0.00\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"225\"\u003e\n\u003cp\u003eSucrose\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"113\"\u003e\n\u003cp\u003e2.00\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"113\"\u003e\n\u003cp\u003e2.00\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"102\"\u003e\n\u003cp\u003e0.00\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"225\"\u003e\n\u003cp\u003eL-lysine (98 %)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"113\"\u003e\n\u003cp\u003e0.40\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"113\"\u003e\n\u003cp\u003e0.40\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"102\"\u003e\n\u003cp\u003e0.40\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"225\"\u003e\n\u003cp\u003eDL- methionine\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"113\"\u003e\n\u003cp\u003e0.11\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"113\"\u003e\n\u003cp\u003e0.11\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"102\"\u003e\n\u003cp\u003e0.11\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"225\"\u003e\n\u003cp\u003eL- threonine\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"113\"\u003e\n\u003cp\u003e0.12\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"113\"\u003e\n\u003cp\u003e0.12\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"102\"\u003e\n\u003cp\u003e0.12\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"225\"\u003e\n\u003cp\u003eL-alanine\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"113\"\u003e\n\u003cp\u003e1.59\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"113\"\u003e\n\u003cp\u003e0.00\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"102\"\u003e\n\u003cp\u003e0.00\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"225\"\u003e\n\u003cp\u003eL-glutamine\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"113\"\u003e\n\u003cp\u003e0.00\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"113\"\u003e\n\u003cp\u003e1.00\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"102\"\u003e\n\u003cp\u003e1.00\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"225\"\u003e\n\u003cp\u003eL-glutamate\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"113\"\u003e\n\u003cp\u003e0.00\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"113\"\u003e\n\u003cp\u003e0.50\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"102\"\u003e\n\u003cp\u003e0.50\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"225\"\u003e\n\u003cp\u003eL-aspartate\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"113\"\u003e\n\u003cp\u003e0.00\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"113\"\u003e\n\u003cp\u003e0.10\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"102\"\u003e\n\u003cp\u003e0.10\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"225\"\u003e\n\u003cp\u003eCarrier\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"113\"\u003e\n\u003cp\u003e0.90\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"113\"\u003e\n\u003cp\u003e0.82\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"102\"\u003e\n\u003cp\u003e0.82\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"225\"\u003e\n\u003cp\u003eOrganic acid calcium\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"113\"\u003e\n\u003cp\u003e0.60\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"113\"\u003e\n\u003cp\u003e0.60\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"102\"\u003e\n\u003cp\u003e0.60\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"225\"\u003e\n\u003cp\u003eCaHPO\u003csub\u003e4\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"113\"\u003e\n\u003cp\u003e1.00\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"113\"\u003e\n\u003cp\u003e1.00\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"102\"\u003e\n\u003cp\u003e1.00\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"225\"\u003e\n\u003cp\u003eCholine chloride (50 %)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"113\"\u003e\n\u003cp\u003e0.01\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"113\"\u003e\n\u003cp\u003e0.01\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"102\"\u003e\n\u003cp\u003e0.01\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"225\"\u003e\n\u003cp\u003eAntioxidant\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"113\"\u003e\n\u003cp\u003e0.05\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"113\"\u003e\n\u003cp\u003e0.05\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"102\"\u003e\n\u003cp\u003e0.05\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"225\"\u003e\n\u003cp\u003eMineral premix \u003csup\u003e2\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"113\"\u003e\n\u003cp\u003e0.15\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"113\"\u003e\n\u003cp\u003e0.15\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"102\"\u003e\n\u003cp\u003e0.15\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"225\"\u003e\n\u003cp\u003eVitamin premix \u003csup\u003e3\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"113\"\u003e\n\u003cp\u003e0.04\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"113\"\u003e\n\u003cp\u003e0.04\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"102\"\u003e\n\u003cp\u003e0.04\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"225\"\u003e\n\u003cp\u003eZnO\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"113\"\u003e\n\u003cp\u003e0.40\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"113\"\u003e\n\u003cp\u003e0.40\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"102\"\u003e\n\u003cp\u003e0.40\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"225\"\u003e\n\u003cp\u003eAcidifier\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"113\"\u003e\n\u003cp\u003e0.70\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"113\"\u003e\n\u003cp\u003e0.70\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"102\"\u003e\n\u003cp\u003e0.70\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"225\"\u003e\n\u003cp\u003eTotal\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"113\"\u003e\n\u003cp\u003e100\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"113\"\u003e\n\u003cp\u003e100\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"102\"\u003e\n\u003cp\u003e100\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd colspan=\"4\" width=\"553\"\u003e\n\u003cp\u003eNutrient composition \u003csup\u003e*\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"225\"\u003e\n\u003cp\u003eDigestible energy (K cal / Kg)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"113\"\u003e\n\u003cp\u003e3445.60\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"113\"\u003e\n\u003cp\u003e3444.56\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"102\"\u003e\n\u003cp\u003e3227.00\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"225\"\u003e\n\u003cp\u003eCrude protein\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"113\"\u003e\n\u003cp\u003e19.57\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"113\"\u003e\n\u003cp\u003e19.53\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"102\"\u003e\n\u003cp\u003e19.53\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"225\"\u003e\n\u003cp\u003eCalcium\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"113\"\u003e\n\u003cp\u003e0.47\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"113\"\u003e\n\u003cp\u003e0.47\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"102\"\u003e\n\u003cp\u003e0.47\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"225\"\u003e\n\u003cp\u003eTotal phosphorus\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"113\"\u003e\n\u003cp\u003e0.40\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"113\"\u003e\n\u003cp\u003e0.40\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"102\"\u003e\n\u003cp\u003e0.40\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"225\"\u003e\n\u003cp\u003eTotal lysine\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"113\"\u003e\n\u003cp\u003e1.14\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"113\"\u003e\n\u003cp\u003e1.14\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"102\"\u003e\n\u003cp\u003e1.11\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u003csup\u003e1 \u003c/sup\u003eT 1 = Basal diet containing 1 % L-glutamine, 0.5 % L-glutamate, and 0.1 % L-aspartate; T 2 = energy deficiency diet containing 1 % L-glutamine, 0.5 % L-glutamate, and 0.1 % L-aspartate; the premix did not contain additional copper, zinc, antibiotics, or probiotics.\u003c/p\u003e\n\u003cp\u003e\u003csup\u003e2 \u003c/sup\u003eMineral premix provided for 1 kg completed diet: Zn (ZnO), 50 mg; Cu (CuSO4), 20 mg; Mn (MnO), 55 mg; Fe (FeSO4), 100 mg; I (KI), 1 mg; Co (CoSO4), 2 mg; Se (Na2SeO3), 0.3 mg.\u003c/p\u003e\n\u003cp\u003e\u003csup\u003e3\u003c/sup\u003e Vitamin premix provided for 1 kg completed diet: vitamin A, 8,255 IU; vitamin D3, 2,000 IU; vitamin E, 40 IU; vitamin B1, 2 mg; vitamin B2, 4 mg; pantothenic acid, 15 mg; vitamin B6, 10 mg; vitamin B12, 0.05 mg; nicotinic acid, 30 mg; folic acid, 2 mg; vitamin K3, 1.5 mg; biotin, 0.2 mg; choline chloride, 800 mg; and vitamin C, 100 mg.\u003c/p\u003e\n\u003cp\u003e\u003csup\u003e* \u003c/sup\u003eCalculated.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 2. Growth performance of weaning piglets\u003csup\u003e1,2\u003c/sup\u003e.\u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"1\" width=\"0\"\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd rowspan=\"2\" width=\"151\"\u003e\n\u003cp\u003eItems\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd colspan=\"3\" width=\"265\"\u003e\n\u003cp\u003eTreatments\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd rowspan=\"2\" width=\"66\"\u003e\n\u003cp\u003eSEM\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd rowspan=\"2\" width=\"66\"\u003e\n\u003cp\u003e\u003cem\u003eP\u003c/em\u003e-value\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"85\"\u003e\n\u003cp\u003eControl\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"95\"\u003e\n\u003cp\u003eT 1\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"85\"\u003e\n\u003cp\u003eT 2\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd colspan=\"6\" width=\"548\"\u003e\n\u003cp\u003eDays 5 post-weaning\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"151\"\u003e\n\u003cp\u003eInitial weight (kg)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"85\"\u003e\n\u003cp\u003e7.09\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"95\"\u003e\n\u003cp\u003e7.05\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"85\"\u003e\n\u003cp\u003e7.09\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"66\"\u003e\n\u003cp\u003e0.070\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"66\"\u003e\n\u003cp\u003e0.959\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"151\"\u003e\n\u003cp\u003eBW on w5d (kg)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"85\"\u003e\n\u003cp\u003e6.89\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"95\"\u003e\n\u003cp\u003e6.91\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"85\"\u003e\n\u003cp\u003e6.94\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"66\"\u003e\n\u003cp\u003e0.063\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"66\"\u003e\n\u003cp\u003e0.952\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"151\"\u003e\n\u003cp\u003eADG (g/pig per day)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"85\"\u003e\n\u003cp\u003e-51.29\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"95\"\u003e\n\u003cp\u003e-34.17\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"85\"\u003e\n\u003cp\u003e-38.83\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"66\"\u003e\n\u003cp\u003e9.767\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"66\"\u003e\n\u003cp\u003e0.782\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"151\"\u003e\n\u003cp\u003eADFI (g/pig per day)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"85\"\u003e\n\u003cp\u003e69.14\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"95\"\u003e\n\u003cp\u003e89.20\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"85\"\u003e\n\u003cp\u003e93.87\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"66\"\u003e\n\u003cp\u003e5.140\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"66\"\u003e\n\u003cp\u003e0.127\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"151\"\u003e\n\u003cp\u003eF/G\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"85\"\u003e\n\u003cp\u003e-1.35\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"95\"\u003e\n\u003cp\u003e-2.61\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"85\"\u003e\n\u003cp\u003e-2.42\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"66\"\u003e\n\u003cp\u003e0.893\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"66\"\u003e\n\u003cp\u003e0.240\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"151\"\u003e\n\u003cp\u003eDigestible energy intake (K cal/pig)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"85\"\u003e\n\u003cp\u003e238.22\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"95\"\u003e\n\u003cp\u003e307.27\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"85\"\u003e\n\u003cp\u003e302.91\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"66\"\u003e\n\u003cp\u003e16.61\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"66\"\u003e\n\u003cp\u003e0.190\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd colspan=\"6\" width=\"548\"\u003e\n\u003cp\u003eDays 21 post-weaning\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"151\"\u003e\n\u003cp\u003eBW on w21d(kg)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"85\"\u003e\n\u003cp\u003e11.58\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"95\"\u003e\n\u003cp\u003e13.17\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"85\"\u003e\n\u003cp\u003e12.07\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"66\"\u003e\n\u003cp\u003e0.278\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"66\"\u003e\n\u003cp\u003e0.047\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"151\"\u003e\n\u003cp\u003eADG (g/pig per day)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"85\"\u003e\n\u003cp\u003e224.57\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"95\"\u003e\n\u003cp\u003e306.00\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"85\"\u003e\n\u003cp\u003e248.88\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"66\"\u003e\n\u003cp\u003e13.801\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"66\"\u003e\n\u003cp\u003e0.035\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"151\"\u003e\n\u003cp\u003eADFI (g/pig per day)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"85\"\u003e\n\u003cp\u003e477.42\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"95\"\u003e\n\u003cp\u003e526.72\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"85\"\u003e\n\u003cp\u003e524.54\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"66\"\u003e\n\u003cp\u003e25.139\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"66\"\u003e\n\u003cp\u003e0.691\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"151\"\u003e\n\u003cp\u003eF/G\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"85\"\u003e\n\u003cp\u003e2.13\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"95\"\u003e\n\u003cp\u003e1.72\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"85\"\u003e\n\u003cp\u003e2.11\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"66\"\u003e\n\u003cp\u003e0.136\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"66\"\u003e\n\u003cp\u003e0.405\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"151\"\u003e\n\u003cp\u003eDigestible energy intake (K cal/pig)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"85\"\u003e\n\u003cp\u003e1644.99\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"95\"\u003e\n\u003cp\u003e1814.33\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"85\"\u003e\n\u003cp\u003e1692.69\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"66\"\u003e\n\u003cp\u003e83.900\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"66\"\u003e\n\u003cp\u003e0.709\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u003csup\u003e1\u003c/sup\u003e Control = basal diets containing 1.59 % L-alanine (iso-nitrogen); T 1 = basal diets containing 1 % glutamine, 0.5 % glutamate, 0.1 % aspartate; T 2 = energy deficiency diets containing 1 % glutamine, 0.5 % glutamate, 0.1 % aspartate; w 5 d = days 5 post-weaning at d 21 of age, w 21 d = days 21 post-weaning at d 21 of age; BW = body weight; ADG = average daily gain; ADFI = average daily feed intake; F / G = gain: feed ratio.\u003c/p\u003e\n\u003cp\u003e\u003csup\u003e2\u003c/sup\u003e \u003csup\u003ea-c \u003c/sup\u003eValues with different letters within the same row are different (\u003cem\u003eP\u003c/em\u003e \u0026lt; 0.05); n = 6.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 3. Jejunal and ileal morphology of weanling piglets \u003csup\u003e1, 2, 3\u003c/sup\u003e. \u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"1\" width=\"100%\"\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd rowspan=\"2\" width=\"16%\"\u003e\n\u003cp\u003eItems\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd colspan=\"3\" width=\"25%\"\u003e\n\u003cp\u003eDiet treatments\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd rowspan=\"2\" width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd colspan=\"2\" width=\"16%\"\u003e\n\u003cp\u003eDays post-weaning\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd rowspan=\"2\" width=\"1%\"\u003e\n\u003cp\u003e\u003cem\u003e\u0026nbsp;\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd colspan=\"2\" width=\"13%\"\u003e\n\u003cp\u003e\u003cem\u003eSEM\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd rowspan=\"2\" width=\"1%\"\u003e\n\u003cp\u003e\u003cem\u003e\u0026nbsp;\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd colspan=\"3\" width=\"23%\"\u003e\n\u003cp\u003e\u003cem\u003eP\u003c/em\u003e-value\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003eControl\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003eT 1\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003eT 2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e5\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e21\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"7%\"\u003e\n\u003cp\u003eDiet\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"5%\"\u003e\n\u003cp\u003eDay\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"6%\"\u003e\n\u003cp\u003eDiet\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"6%\"\u003e\n\u003cp\u003eDay\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"9%\"\u003e\n\u003cp\u003eDiet \u0026times; Day\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd colspan=\"14\" width=\"100%\"\u003e\n\u003cp\u003eJejunum\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"16%\"\u003e\n\u003cp\u003eVillus height (\u0026mu;m)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e375.31\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e393.77\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e392.16\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e337.64\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e436.52\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"7%\"\u003e\n\u003cp\u003e10.44\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"5%\"\u003e\n\u003cp\u003e8.52\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"6%\"\u003e\n\u003cp\u003e0.394\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"6%\"\u003e\n\u003cp\u003e\u0026lt;0.001\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"9%\"\u003e\n\u003cp\u003e0.049\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"16%\"\u003e\n\u003cp\u003eCrypt depth (\u0026mu;m)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e119.98\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e117.11\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e118.58\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e112.95\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e124.16\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"7%\"\u003e\n\u003cp\u003e5.15\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"5%\"\u003e\n\u003cp\u003e4.20\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"6%\"\u003e\n\u003cp\u003e0.925\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"6%\"\u003e\n\u003cp\u003e0.069\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"9%\"\u003e\n\u003cp\u003e0.284\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"16%\"\u003e\n\u003cp\u003eVCR\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e3.15\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e3.43\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e3.36\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e3.02\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e3.61\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"7%\"\u003e\n\u003cp\u003e0.17\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"5%\"\u003e\n\u003cp\u003e0.14\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"6%\"\u003e\n\u003cp\u003e0.483\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"6%\"\u003e\n\u003cp\u003e0.006\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"9%\"\u003e\n\u003cp\u003e0.815\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"16%\"\u003e\n\u003cp\u003eGoblet cells\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e10.68\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e10.23\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e11.17\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e10.42\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e10.97\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"7%\"\u003e\n\u003cp\u003e0.25\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"5%\"\u003e\n\u003cp\u003e0.20\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"6%\"\u003e\n\u003cp\u003e0.044\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"6%\"\u003e\n\u003cp\u003e0.069\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"9%\"\u003e\n\u003cp\u003e0.503\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"16%\"\u003e\n\u003cp\u003eLymphocyte cells\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e38.98\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e38.7\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e39.62\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e39.73\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e38.47\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"7%\"\u003e\n\u003cp\u003e0.58\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"5%\"\u003e\n\u003cp\u003e0.47\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"6%\"\u003e\n\u003cp\u003e0.528\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"6%\"\u003e\n\u003cp\u003e0.069\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"9%\"\u003e\n\u003cp\u003e0.626\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd colspan=\"14\" width=\"100%\"\u003e\n\u003cp\u003eIleum\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"16%\"\u003e\n\u003cp\u003eVillus height (\u0026mu;m)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e358.17\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e390.65\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e347.49\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e322.49\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e408.39\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"7%\"\u003e\n\u003cp\u003e10.21\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"5%\"\u003e\n\u003cp\u003e8.34\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"6%\"\u003e\n\u003cp\u003e0.015\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"6%\"\u003e\n\u003cp\u003e\u0026lt;0.001\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"9%\"\u003e\n\u003cp\u003e0.020\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"16%\"\u003e\n\u003cp\u003eCrypt depth (\u0026mu;m)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e118.32\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e117.98\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e106.56\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e101.34\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e127.23\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"7%\"\u003e\n\u003cp\u003e3.48\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"5%\"\u003e\n\u003cp\u003e2.84\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"6%\"\u003e\n\u003cp\u003e0.037\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"6%\"\u003e\n\u003cp\u003e\u0026lt;0.001\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"9%\"\u003e\n\u003cp\u003e0.046\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"16%\"\u003e\n\u003cp\u003eVCR\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e3.05\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e3.37\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e3.26\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e3.22\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e3.23\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"7%\"\u003e\n\u003cp\u003e0.13\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"5%\"\u003e\n\u003cp\u003e0.10\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"6%\"\u003e\n\u003cp\u003e0.203\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"6%\"\u003e\n\u003cp\u003e0.936\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"9%\"\u003e\n\u003cp\u003e0.634\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"16%\"\u003e\n\u003cp\u003eGoblet cells\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e10.53\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e10.82\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e10.72\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e10.53\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e10.84\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"7%\"\u003e\n\u003cp\u003e0.32\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"5%\"\u003e\n\u003cp\u003e0.26\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"6%\"\u003e\n\u003cp\u003e0.816\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"6%\"\u003e\n\u003cp\u003e0.403\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"9%\"\u003e\n\u003cp\u003e0.455\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"16%\"\u003e\n\u003cp\u003eLymphocyte cells\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e40.12\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e40.67\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e39.65\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e40.03\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e40.26\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"7%\"\u003e\n\u003cp\u003e0.38\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"5%\"\u003e\n\u003cp\u003e0.31\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"6%\"\u003e\n\u003cp\u003e0.189\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"6%\"\u003e\n\u003cp\u003e0.619\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"9%\"\u003e\n\u003cp\u003e0.483\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u003csup\u003e1\u003c/sup\u003e Control = basal diets containing 1.59 % L-alanine (iso-nitrogen); T 1 = basal diets containing 1 % glutamine, 0.5 % glutamate, 0.1 % aspartate; T 2 = energy deficiency diets containing 1 % glutamine, 0.5 % glutamate, 0.1 % aspartate; w 5 d = days 5 post-weaning at d 21 of age, w 21 d = days 21 post-weaning at d 21 of age; VCR = the ratio of villus height to crypt depth.\u003c/p\u003e\n\u003cp\u003e\u003csup\u003e2\u003c/sup\u003e The unit of goblet cells and lymphocyte is cell number per area.\u003c/p\u003e\n\u003cp\u003e\u003csup\u003e3\u003c/sup\u003e The results were presented as main effects, and the differences among treatments or days were evaluated using Tukey HSD test. \u003csup\u003ea-b \u003c/sup\u003eValues with different letters within the same row are different (\u003cem\u003eP\u003c/em\u003e \u0026lt; 0.05); n = 6.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable S1. Primers used for real-time quantitative reverse transcription-PCR \u003csup\u003e1\u003c/sup\u003e.\u003c/strong\u003e\u003c/p\u003e\n\u003ctable style=\"width: 67.4875%;\" border=\"1\"\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 13%;\"\u003e\n\u003cp\u003eGene\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 25%;\"\u003e\n\u003cp\u003eAccession No.\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 27.6195%;\"\u003e\n\u003cp\u003ePrimers\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 13%;\" rowspan=\"2\"\u003e\n\u003cp\u003e\u003cem\u003e\u0026beta;\u003c/em\u003e\u003cem\u003e-Actin\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 25%;\" rowspan=\"2\"\u003e\n\u003cp\u003eXM_003124280.3\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 27.6195%;\"\u003e\n\u003cp\u003eF: 5\u0026rsquo;-GGATGCAGAAGGAGATCACG-3\u0026rsquo;\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 27.6195%;\"\u003e\n\u003cp\u003eR: 5\u0026rsquo;-ATCTGCTGGAAGGTGGACAG-3\u0026rsquo;\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 13%;\" rowspan=\"2\"\u003e\n\u003cp\u003e\u003cem\u003ePdk4 \u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 25%;\" rowspan=\"2\"\u003e\n\u003cp\u003eNM_001159306.1\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 27.6195%;\"\u003e\n\u003cp\u003eF: 5\u0026rsquo;-CTACAGCACCAACACCTGTGA-3\u0026rsquo;\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 27.6195%;\"\u003e\n\u003cp\u003eR: 5\u0026rsquo;-CATCCGTTCCATATCCTGGCA-3\u0026rsquo;\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 13%;\" rowspan=\"2\"\u003e\n\u003cp\u003e\u003cem\u003ePck1\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 25%;\" rowspan=\"2\"\u003e\n\u003cp\u003eXM_005673043.3\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 27.6195%;\"\u003e\n\u003cp\u003eF: 5\u0026rsquo;-TCTGGTGTACGAGGCTCTCA-3\u0026rsquo;\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 27.6195%;\"\u003e\n\u003cp\u003eR: 5\u0026rsquo;-TTGCCGAAGTTGTAGCCGAA-3\u0026rsquo;\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 13%;\" rowspan=\"2\"\u003e\n\u003cp\u003e\u003cem\u003eSdh1 \u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 25%;\" rowspan=\"2\"\u003e\n\u003cp\u003eNM_001179714.1\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 27.6195%;\"\u003e\n\u003cp\u003eF: 5\u0026rsquo;-CCGCTGGTCTCTTCCTATGG-3\u0026rsquo;\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 27.6195%;\"\u003e\n\u003cp\u003eR: 5\u0026rsquo;-TGGCCCTAAACACAGGGACT-3\u0026rsquo;\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 13%;\" rowspan=\"2\"\u003e\n\u003cp\u003e\u003cem\u003eUcp2 \u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 25%;\" rowspan=\"2\"\u003e\n\u003cp\u003eNM_214289.1\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 27.6195%;\"\u003e\n\u003cp\u003eF: 5\u0026rsquo;-CTCACCCAATGTCGCTCGTA-3\u0026rsquo;\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 27.6195%;\"\u003e\n\u003cp\u003eR: 5\u0026rsquo;-ATCTCGTCTTGACCACGTCC-3\u0026rsquo;\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 13%;\" rowspan=\"2\"\u003e\n\u003cp\u003e\u003cem\u003ePpar\u003c/em\u003e\u003cem\u003e\u0026alpha; 1\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 25%;\" rowspan=\"2\"\u003e\n\u003cp\u003eNM_001044526.1\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 27.6195%;\"\u003e\n\u003cp\u003eF: 5\u0026rsquo;-GCAGATCTCAAGTCTCTCGCC-3\u0026rsquo;\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 27.6195%;\"\u003e\n\u003cp\u003eR: 5\u0026rsquo;-CCTTGTTCTGGATGCCGTTG-3\u0026rsquo;\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 13%;\" rowspan=\"2\"\u003e\n\u003cp\u003e\u003cem\u003eAcox1\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 25%;\" rowspan=\"2\"\u003e\n\u003cp\u003eNM_001101028.1\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 27.6195%;\"\u003e\n\u003cp\u003eF: 5\u0026rsquo;-TGTCAACTTCACCCCAACCTG -3\u0026rsquo;\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 27.6195%;\"\u003e\n\u003cp\u003eR: 5\u0026rsquo;-AGGTAGGACACCATGCCACA -3\u0026rsquo;\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 13%;\" rowspan=\"2\"\u003e\n\u003cp\u003e\u003cem\u003eCpt1\u003c/em\u003e\u003cem\u003e\u0026alpha;\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 25%;\" rowspan=\"2\"\u003e\n\u003cp\u003eNM_001129805.1\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 27.6195%;\"\u003e\n\u003cp\u003eF: 5\u0026rsquo;-GCATTTGTCCCATCTTTCGT -3\u0026rsquo;\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 27.6195%;\"\u003e\n\u003cp\u003eR: 5\u0026rsquo;-GCACTGGTCCTTCTGGGATA -3\u0026rsquo;\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 13%;\" rowspan=\"2\"\u003e\n\u003cp\u003e\u003cem\u003eAmpk\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 25%;\" rowspan=\"2\"\u003e\n\u003cp\u003eXM_021076522.1\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 27.6195%;\"\u003e\n\u003cp\u003eF: 5\u0026rsquo;-CTGGATTATGAGTGGAAGGT-3\u0026rsquo;\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 27.6195%;\"\u003e\n\u003cp\u003eR: 5\u0026rsquo;-AAGTCAACAGGAGAAGAGTC-3\u0026rsquo;\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 13%;\" rowspan=\"2\"\u003e\n\u003cp\u003e\u003cem\u003eAcc\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 25%;\"\u003e\n\u003cp\u003eNM_001114269.1\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 27.6195%;\"\u003e\n\u003cp\u003eF: 5\u0026rsquo;-GCCGAAACATCTCTGGGATA -3\u0026rsquo;\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 25%;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 27.6195%;\"\u003e\n\u003cp\u003eR: 5\u0026rsquo;-CTCCAGGACAGCACAGATCA -3\u0026rsquo;\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 13%;\" rowspan=\"2\"\u003e\n\u003cp\u003e\u003cem\u003eSirt1\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 25%;\" rowspan=\"2\"\u003e\n\u003cp\u003eNM_001145750.2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 27.6195%;\"\u003e\n\u003cp\u003eF: 5\u0026rsquo;-AACATGAAGAGGTGTGGGTG -3\u0026rsquo;\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 27.6195%;\"\u003e\n\u003cp\u003eR: 5\u0026rsquo;-GAACAGGTTGCGGGAATC -3\u0026rsquo;\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 13%;\" rowspan=\"2\"\u003e\n\u003cp\u003e\u003cem\u003ePgc1\u003c/em\u003e\u003cem\u003e\u0026alpha;\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 25%;\" rowspan=\"2\"\u003e\n\u003cp\u003eXM_005665970.3\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 27.6195%;\"\u003e\n\u003cp\u003eF: 5\u0026rsquo;-GTGAGACAGAGCCAGGTA-3\u0026rsquo;\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 27.6195%;\"\u003e\n\u003cp\u003eR: 5\u0026rsquo;-TTGATGATGAAGGTGAAGGT-3\u0026rsquo;\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 13%;\" rowspan=\"2\"\u003e\n\u003cp\u003e\u003cem\u003eTfam\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 25%;\" rowspan=\"2\"\u003e\n\u003cp\u003eNM_001130211.1\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 27.6195%;\"\u003e\n\u003cp\u003eF: 5\u0026rsquo;-CCACCCTGAGTGGTTTTCCA-3\u0026rsquo;\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 27.6195%;\"\u003e\n\u003cp\u003eR: 5\u0026rsquo;-TGCCAGTCTGCCCTATAAGC-3\u0026rsquo;\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 13%;\" rowspan=\"2\"\u003e\n\u003cp\u003e\u003cem\u003eNrf1\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 25%;\" rowspan=\"2\"\u003e\n\u003cp\u003eXM_021078993.1\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 27.6195%;\"\u003e\n\u003cp\u003eF: 5\u0026rsquo;-CATGGCACTCAACAGCGAAG-3\u0026rsquo;\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 27.6195%;\"\u003e\n\u003cp\u003eR: 5\u0026rsquo;-ACATGCTCACAGGGATCTGG-3\u0026rsquo;\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u003csup\u003e1 \u003c/sup\u003ePdk4: pyruvate dehydrogenase kinase, Pck1: phosphoenolpyruvate carboxykinase 1, Sdh1: succinate dehydrogenase 1, Ucp2: mitochondrial uncoupling protein 2, Ppar\u0026alpha;1: peroxisome proliferator-activated receptor alpha 1, Acox1: peroxisomal acyl-coenzyme A oxidase 1, Cpt1\u0026alpha;: carnitine palmitoyltransferase 1A, Ampk, AMP-activated protein kinase, Acc: acetyl-CoA carboxylase, Sirt1: Sirtuin 1, Pgc1\u003cem\u003e\u0026alpha;\u003c/em\u003e: peroxisome proliferator-activated receptor gamma coactivator 1-\u0026alpha;, Nrf1: nuclear respiratory factor 1, Tfam: mitochondrial transcription factor A. F: forward; R: reverse.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable S2. The relative density of jejunal mucosal metabolites of weanling piglets \u003c/strong\u003e\u003cstrong\u003e\u003csup\u003e1, 2\u003c/sup\u003e\u003c/strong\u003e\u003cstrong\u003e.\u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"1\" width=\"100%\"\u003e\n\u003ctbody\u003e\n\u003ctr style=\"height: 35.4287px;\"\u003e\n\u003ctd style=\"height: 70.4287px; width: 16%;\" rowspan=\"2\"\u003e\n\u003cp\u003eItems\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35.4287px; width: 26.0439%;\" colspan=\"3\"\u003e\n\u003cp\u003eDiet treatments\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 70.4287px; width: 1%;\" rowspan=\"2\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35.4287px; width: 16%;\" colspan=\"2\"\u003e\n\u003cp\u003eDays post-weaning\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 70.4287px; width: 1%;\" rowspan=\"2\"\u003e\n\u003cp\u003e\u003cem\u003e\u0026nbsp;\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35.4287px; width: 12%;\" colspan=\"2\"\u003e\n\u003cp\u003e\u003cem\u003eSEM\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 70.4287px; width: 1%;\" rowspan=\"2\"\u003e\n\u003cp\u003e\u003cem\u003e\u0026nbsp;\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35.4287px; width: 21%;\" colspan=\"3\"\u003e\n\u003cp\u003e\u003cem\u003eP\u003c/em\u003e-value\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr style=\"height: 35px;\"\u003e\n\u003ctd style=\"height: 35px; width: 8%;\"\u003e\n\u003cp\u003eControl\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 8.04388%;\"\u003e\n\u003cp\u003eT 1\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 10%;\"\u003e\n\u003cp\u003eT 2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 8%;\"\u003e\n\u003cp\u003e5\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 8%;\"\u003e\n\u003cp\u003e21\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 7%;\"\u003e\n\u003cp\u003eDiet\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 5%;\"\u003e\n\u003cp\u003eDay\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 6%;\"\u003e\n\u003cp\u003eDiet\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 6%;\"\u003e\n\u003cp\u003eDay\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 9%;\"\u003e\n\u003cp\u003eDiet \u0026times; Day\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr style=\"height: 35px;\"\u003e\n\u003ctd style=\"height: 35px; width: 16%;\"\u003e\n\u003cp\u003eAKG\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 8%;\"\u003e\n\u003cp\u003e1.00\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 8.04388%;\"\u003e\n\u003cp\u003e1.22\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 10%;\"\u003e\n\u003cp\u003e1.09\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 1%;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 8%;\"\u003e\n\u003cp\u003e1.21\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 8%;\"\u003e\n\u003cp\u003e0.99\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 1%;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 7%;\"\u003e\n\u003cp\u003e0.03\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 5%;\"\u003e\n\u003cp\u003e0.02\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 1%;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 6%;\"\u003e\n\u003cp\u003e\u0026lt;0.001\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 6%;\"\u003e\n\u003cp\u003e\u0026lt;0.001\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 9%;\"\u003e\n\u003cp\u003e\u0026lt;0.001\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr style=\"height: 35px;\"\u003e\n\u003ctd style=\"height: 35px; width: 16%;\"\u003e\n\u003cp\u003eOxaloacetate\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 8%;\"\u003e\n\u003cp\u003e1.00\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 8.04388%;\"\u003e\n\u003cp\u003e1.08\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 10%;\"\u003e\n\u003cp\u003e1.15\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 1%;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 8%;\"\u003e\n\u003cp\u003e0.96\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 8%;\"\u003e\n\u003cp\u003e1.20\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 1%;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 7%;\"\u003e\n\u003cp\u003e0.07\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 5%;\"\u003e\n\u003cp\u003e0.06\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 1%;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 6%;\"\u003e\n\u003cp\u003e0.307\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 6%;\"\u003e\n\u003cp\u003e0.005\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 9%;\"\u003e\n\u003cp\u003e0.047\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr style=\"height: 38px;\"\u003e\n\u003ctd style=\"height: 38px; width: 16%;\"\u003e\n\u003cp\u003eSuccinate\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 38px; width: 8%;\"\u003e\n\u003cp\u003e1.00\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 38px; width: 8.04388%;\"\u003e\n\u003cp\u003e1.49\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 38px; width: 10%;\"\u003e\n\u003cp\u003e1.33\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 38px; width: 1%;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 38px; width: 8%;\"\u003e\n\u003cp\u003e1.31\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 38px; width: 8%;\"\u003e\n\u003cp\u003e1.24\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 38px; width: 1%;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 38px; width: 7%;\"\u003e\n\u003cp\u003e0.10\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 38px; width: 5%;\"\u003e\n\u003cp\u003e0.08\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 38px; width: 1%;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 38px; width: 6%;\"\u003e\n\u003cp\u003e0.005\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 38px; width: 6%;\"\u003e\n\u003cp\u003e0.572\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 38px; width: 9%;\"\u003e\n\u003cp\u003e0.091\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr style=\"height: 38px;\"\u003e\n\u003ctd style=\"height: 38px; width: 16%;\"\u003e\n\u003cp\u003ePhosphoenolpyruvate\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 38px; width: 8%;\"\u003e\n\u003cp\u003e1.00\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 38px; width: 8.04388%;\"\u003e\n\u003cp\u003e0.55\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 38px; width: 10%;\"\u003e\n\u003cp\u003e0.40\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 38px; width: 1%;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 38px; width: 8%;\"\u003e\n\u003cp\u003e0.68\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 38px; width: 8%;\"\u003e\n\u003cp\u003e0.62\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 38px; width: 1%;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 38px; width: 7%;\"\u003e\n\u003cp\u003e0.11\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 38px; width: 5%;\"\u003e\n\u003cp\u003e0.09\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 38px; 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width: 1%;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 8%;\"\u003e\n\u003cp\u003e1.05\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 8%;\"\u003e\n\u003cp\u003e0.98\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 1%;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 7%;\"\u003e\n\u003cp\u003e0.05\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 5%;\"\u003e\n\u003cp\u003e0.04\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 1%;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 6%;\"\u003e\n\u003cp\u003e0.897\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 6%;\"\u003e\n\u003cp\u003e0.159\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 9%;\"\u003e\n\u003cp\u003e0.546\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr style=\"height: 35px;\"\u003e\n\u003ctd style=\"height: 35px; width: 16%;\"\u003e\n\u003cp\u003eIsocitrate\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 8%;\"\u003e\n\u003cp\u003e1.00\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 8.04388%;\"\u003e\n\u003cp\u003e1.34\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 10%;\"\u003e\n\u003cp\u003e1.33\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 1%;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 8%;\"\u003e\n\u003cp\u003e1.45\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 8%;\"\u003e\n\u003cp\u003e1.00\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 1%;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 7%;\"\u003e\n\u003cp\u003e0.08\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 5%;\"\u003e\n\u003cp\u003e0.07\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; 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width: 8%;\"\u003e\n\u003cp\u003e1.03\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 8%;\"\u003e\n\u003cp\u003e1.10\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 1%;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 7%;\"\u003e\n\u003cp\u003e0.16\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 5%;\"\u003e\n\u003cp\u003e0.13\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 1%;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 6%;\"\u003e\n\u003cp\u003e0.831\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 6%;\"\u003e\n\u003cp\u003e0.714\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 9%;\"\u003e\n\u003cp\u003e0.508\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr style=\"height: 35px;\"\u003e\n\u003ctd style=\"height: 35px; width: 16%;\"\u003e\n\u003cp\u003ePyruvate\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 8%;\"\u003e\n\u003cp\u003e1.00\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 8.04388%;\"\u003e\n\u003cp\u003e1.20\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 10%;\"\u003e\n\u003cp\u003e1.17\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 1%;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 8%;\"\u003e\n\u003cp\u003e1.24\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 8%;\"\u003e\n\u003cp\u003e1.01\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 1%;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 7%;\"\u003e\n\u003cp\u003e0.06\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 5%;\"\u003e\n\u003cp\u003e0.05\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 1%;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; 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width: 5%;\"\u003e\n\u003cp\u003e0.08\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 1%;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 6%;\"\u003e\n\u003cp\u003e0.870\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 6%;\"\u003e\n\u003cp\u003e0.004\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 9%;\"\u003e\n\u003cp\u003e0.017\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr style=\"height: 35px;\"\u003e\n\u003ctd style=\"height: 35px; width: 16%;\"\u003e\n\u003cp\u003e3-phosphoglycerate\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 8%;\"\u003e\n\u003cp\u003e1.00\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 8.04388%;\"\u003e\n\u003cp\u003e0.87\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 10%;\"\u003e\n\u003cp\u003e0.69\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 1%;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 8%;\"\u003e\n\u003cp\u003e0.83\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 8%;\"\u003e\n\u003cp\u003e0.87\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 1%;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 7%;\"\u003e\n\u003cp\u003e0.07\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 5%;\"\u003e\n\u003cp\u003e0.06\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 1%;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 6%;\"\u003e\n\u003cp\u003e0.013\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 6%;\"\u003e\n\u003cp\u003e0.605\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 9%;\"\u003e\n\u003cp\u003e0.001\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr style=\"height: 35px;\"\u003e\n\u003ctd style=\"height: 35px; width: 16%;\"\u003e\n\u003cp\u003eG-6-P\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 8%;\"\u003e\n\u003cp\u003e1.00\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 8.04388%;\"\u003e\n\u003cp\u003e0.99\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 10%;\"\u003e\n\u003cp\u003e1.11\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 1%;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 8%;\"\u003e\n\u003cp\u003e0.87\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 8%;\"\u003e\n\u003cp\u003e1.19\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 1%;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 7%;\"\u003e\n\u003cp\u003e0.13\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 5%;\"\u003e\n\u003cp\u003e0.11\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 1%;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 6%;\"\u003e\n\u003cp\u003e0.749\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 6%;\"\u003e\n\u003cp\u003e0.041\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 9%;\"\u003e\n\u003cp\u003e0.009\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr style=\"height: 35px;\"\u003e\n\u003ctd style=\"height: 35px; width: 16%;\"\u003e\n\u003cp\u003eDHAP\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 8%;\"\u003e\n\u003cp\u003e1.00\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 8.04388%;\"\u003e\n\u003cp\u003e0.81\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 10%;\"\u003e\n\u003cp\u003e1.53\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 1%;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 8%;\"\u003e\n\u003cp\u003e1.36\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 8%;\"\u003e\n\u003cp\u003e0.86\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 1%;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 7%;\"\u003e\n\u003cp\u003e0.09\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 5%;\"\u003e\n\u003cp\u003e0.08\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 1%;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 6%;\"\u003e\n\u003cp\u003e\u0026lt;0.001\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 6%;\"\u003e\n\u003cp\u003e\u0026lt;0.001\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"height: 35px; width: 9%;\"\u003e\n\u003cp\u003e\u0026lt;0.001\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u003csup\u003e1\u003c/sup\u003e Control, basal diets containing 1.59 % L-alanine (iso-nitrogen); T1, basal diets containing 1 % glutamine, 0.5 % glutamate, 0.1 % aspartate; T2, energy deficiency diets containing 1 % glutamine, 0.5 % glutamate, 0.1 % aspartate. G-6-P: glucose-6-phosphate, F-6-P: fructose 6 phosphate, F-1,6-P: fructose 1,6 phosphate, DHAP: dihydroxyacetone phosphate, AKG: \u0026alpha;-ketoglutarate, ATP: adenosine 5 triphosphate, ADP: adenosine 5 diphosphate, AMP: adenosine 5 monophosphate, GTP: guanosine 5 triphosphate, GDP: guanosine 5 diphosphate, GMP: guanosine 5 monophosphate, NAD\u003csup\u003e+\u003c/sup\u003e: nicotinamide adenine dinucleotide\u003csup\u003e+\u003c/sup\u003e, NADH: nicotinamide adenine dinucleotide hydrogen, NADP\u003csup\u003e+\u003c/sup\u003e: nicotinamide adenine dinucleotide phosphate, NADPH: nicotinamide adenine dinucleotide phosphate hydrogen, TPP: thiamine pyrophosphate, FMN: flavin mononucleotide.\u003c/p\u003e\n\u003cp\u003e\u003csup\u003e2\u003c/sup\u003e The results were presented as main effects, and the differences among treatments or days were evaluated using Tukey HSD test. \u003csup\u003ea-b \u003c/sup\u003eValues with different letters within the same row are different (\u003cem\u003eP\u003c/em\u003e \u0026lt; 0.05); n = 6.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable S3. Relative expressions of gene involved in the energy metabolism\u003csup\u003e1,2\u003c/sup\u003e. \u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"1\" width=\"100%\"\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd rowspan=\"2\" width=\"16%\"\u003e\n\u003cp\u003eItems\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd colspan=\"3\" width=\"25%\"\u003e\n\u003cp\u003eDiet treatments\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd rowspan=\"2\" width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd colspan=\"2\" width=\"16%\"\u003e\n\u003cp\u003eDays post-weaning\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd rowspan=\"2\" width=\"1%\"\u003e\n\u003cp\u003e\u003cem\u003e\u0026nbsp;\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd colspan=\"2\" width=\"13%\"\u003e\n\u003cp\u003e\u003cem\u003eSEM\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd rowspan=\"2\" width=\"1%\"\u003e\n\u003cp\u003e\u003cem\u003e\u0026nbsp;\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd colspan=\"3\" width=\"23%\"\u003e\n\u003cp\u003e\u003cem\u003eP\u003c/em\u003e-value\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003eControl\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003eT 1\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003eT 2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e5\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e21\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"7%\"\u003e\n\u003cp\u003eDiet\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"5%\"\u003e\n\u003cp\u003eDay\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"6%\"\u003e\n\u003cp\u003eDiet\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"6%\"\u003e\n\u003cp\u003eDay\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"9%\"\u003e\n\u003cp\u003eDiet \u0026times; Day\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd colspan=\"14\" width=\"100%\"\u003e\n\u003cp\u003eJejunal mucosa\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"16%\"\u003e\n\u003cp\u003e\u003cem\u003ePdk4\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e0.99\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e1.31\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e0.79\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e0.95\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e1.11\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"7%\"\u003e\n\u003cp\u003e0.15\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"5%\"\u003e\n\u003cp\u003e0.13\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"6%\"\u003e\n\u003cp\u003e0.089\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"6%\"\u003e\n\u003cp\u003e0.402\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"9%\"\u003e\n\u003cp\u003e0.009\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"16%\"\u003e\n\u003cp\u003e\u003cem\u003ePck1\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e1.00\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e0.76\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e0.97\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e0.77\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e1.05\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"7%\"\u003e\n\u003cp\u003e0.28\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"5%\"\u003e\n\u003cp\u003e0.24\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"6%\"\u003e\n\u003cp\u003e0.822\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"6%\"\u003e\n\u003cp\u003e0.399\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"9%\"\u003e\n\u003cp\u003e0.038\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"16%\"\u003e\n\u003cp\u003e\u003cem\u003eSdh1\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e1.09\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e2.03\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e2.01\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e0.88\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e2.54\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"7%\"\u003e\n\u003cp\u003e0.33\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"5%\"\u003e\n\u003cp\u003e0.27\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"6%\"\u003e\n\u003cp\u003e0.085\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"6%\"\u003e\n\u003cp\u003e\u0026lt;0.001\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"9%\"\u003e\n\u003cp\u003e0.007\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"16%\"\u003e\n\u003cp\u003e\u003cem\u003eUcp2\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e1.00\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e1.02\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e1.92\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e1.54\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e1.08\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"7%\"\u003e\n\u003cp\u003e0.54\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"5%\"\u003e\n\u003cp\u003e0.43\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"6%\"\u003e\n\u003cp\u003e0.393\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"6%\"\u003e\n\u003cp\u003e0.466\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"9%\"\u003e\n\u003cp\u003e0.407\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"16%\"\u003e\n\u003cp\u003e\u003cem\u003ePpar\u0026alpha;1\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e0.91\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e1.25\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e1.91\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e1.95\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e0.76\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd 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width=\"8%\"\u003e\n\u003cp\u003e1.10\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"7%\"\u003e\n\u003cp\u003e0.21\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"5%\"\u003e\n\u003cp\u003e0.17\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"6%\"\u003e\n\u003cp\u003e0.640\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"6%\"\u003e\n\u003cp\u003e0.928\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"9%\"\u003e\n\u003cp\u003e0.98\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"16%\"\u003e\n\u003cp\u003e\u003cem\u003eAcox1\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e1.00\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e1.30\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e1.38\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e1.14\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e1.31\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"7%\"\u003e\n\u003cp\u003e0.21\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"5%\"\u003e\n\u003cp\u003e0.17\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"6%\"\u003e\n\u003cp\u003e0.415\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"6%\"\u003e\n\u003cp\u003e0.502\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"9%\"\u003e\n\u003cp\u003e0.889\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"16%\"\u003e\n\u003cp\u003e\u003cem\u003eCpt1\u0026alpha;\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e1.00\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e0.81\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e1.39\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e1.13\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e1.01\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"7%\"\u003e\n\u003cp\u003e0.05\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"5%\"\u003e\n\u003cp\u003e0.04\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"6%\"\u003e\n\u003cp\u003e\u0026lt;0.001\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"6%\"\u003e\n\u003cp\u003e0.069\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"9%\"\u003e\n\u003cp\u003e0.063\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u003csup\u003e1\u003c/sup\u003e Control, basal diets containing 1.59 % L-alanine (iso-nitrogen); T1, basal diets containing 1 % glutamine, 0.5 % glutamate, 0.1 % aspartate; T2, energy deficiency diets containing 1 % glutamine, 0.5 % glutamate, 0.1 % aspartate. Pdk4: pyruvate dehydrogenase kinase, Pck1: phosphoenolpyruvate carboxykinase 1, Sdh1: succinate dehydrogenase 1, Ucp2: mitochondrial uncoupling protein 2, Ppar\u0026alpha;1: peroxisome proliferator-activated receptor alpha 1, Acox1: peroxisomal acyl-coenzyme A oxidase 1, Cpt1\u0026alpha;: carnitine palmitoyltransferase 1A, ACC: acetyl-CoA carboxylase, SirT1: Sirtuin 1, PGC1\u003cem\u003e\u0026alpha;\u003c/em\u003e: peroxisome proliferator-activated receptor gamma coactivator 1-\u0026alpha;, Nrf1: nuclear respiratory factor 1, TFAM: mitochondrial transcription factor A.\u003c/p\u003e\n\u003cp\u003e\u003csup\u003e2\u003c/sup\u003e The results were presented as main effects, and the differences among treatments or days were evaluated using Tukey HSD test. \u003csup\u003ea-b \u003c/sup\u003eValues with different letters within the same row are different (\u003cem\u003eP\u003c/em\u003e \u0026lt; 0.05); n = 6.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable S4. Relative gene expressions of AMPK signaling pathway of jejunal and ileal mucosa of weanling piglets \u003csup\u003e1,2\u003c/sup\u003e.\u0026nbsp; \u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"1\" width=\"100%\"\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd rowspan=\"2\" width=\"16%\"\u003e\n\u003cp\u003eItems\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd colspan=\"3\" width=\"25%\"\u003e\n\u003cp\u003eDiet treatments\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd rowspan=\"2\" width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd colspan=\"2\" width=\"16%\"\u003e\n\u003cp\u003eDays post-weaning\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd rowspan=\"2\" width=\"1%\"\u003e\n\u003cp\u003e\u003cem\u003e\u0026nbsp;\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd colspan=\"2\" width=\"13%\"\u003e\n\u003cp\u003e\u003cem\u003eSEM\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd rowspan=\"2\" width=\"1%\"\u003e\n\u003cp\u003e\u003cem\u003e\u0026nbsp;\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd colspan=\"3\" width=\"23%\"\u003e\n\u003cp\u003e\u003cem\u003eP\u003c/em\u003e-value\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003eControl\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003eT 1\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003eT 2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e5\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e21\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"7%\"\u003e\n\u003cp\u003eDiet\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"5%\"\u003e\n\u003cp\u003eDay\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"6%\"\u003e\n\u003cp\u003eDiet\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"6%\"\u003e\n\u003cp\u003eDay\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"9%\"\u003e\n\u003cp\u003eDiet \u0026times; Day\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd colspan=\"14\" width=\"100%\"\u003e\n\u003cp\u003eJejunal mucosa\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"16%\"\u003e\n\u003cp\u003e\u003cem\u003eAmpk\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e1.00\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e0.87\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e1.05\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e0.64\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e1.39\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"7%\"\u003e\n\u003cp\u003e0.30\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"5%\"\u003e\n\u003cp\u003e0.31\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"6%\"\u003e\n\u003cp\u003e0.893\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"6%\"\u003e\n\u003cp\u003e0.048\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"9%\"\u003e\n\u003cp\u003e0.364\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"16%\"\u003e\n\u003cp\u003e\u003cem\u003eAcc\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e1.00\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e0.99\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e1.01\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e0.58\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e1.41\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"7%\"\u003e\n\u003cp\u003e0.32\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"5%\"\u003e\n\u003cp\u003e0.26\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"6%\"\u003e\n\u003cp\u003e0.999\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"6%\"\u003e\n\u003cp\u003e0.032\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"9%\"\u003e\n\u003cp\u003e0.281\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"16%\"\u003e\n\u003cp\u003e\u003cem\u003eSirt1\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e1.04\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e0.99\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e1.59\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e1.45\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e0.96\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"7%\"\u003e\n\u003cp\u003e0.39\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"5%\"\u003e\n\u003cp\u003e0.32\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"6%\"\u003e\n\u003cp\u003e0.493\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"6%\"\u003e\n\u003cp\u003e0.282\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"9%\"\u003e\n\u003cp\u003e0.449\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"16%\"\u003e\n\u003cp\u003e\u003cem\u003ePgc1\u0026alpha;\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e0.99\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e0.37\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e0.86\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e0.55\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e0.89\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"7%\"\u003e\n\u003cp\u003e0.14\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"5%\"\u003e\n\u003cp\u003e0.11\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"6%\"\u003e\n\u003cp\u003e0.013\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"6%\"\u003e\n\u003cp\u003e0.038\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"9%\"\u003e\n\u003cp\u003e0.105\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"16%\"\u003e\n\u003cp\u003e\u003cem\u003eTfam\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e1.00\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e0.49\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e0.78\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e0.69\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e0.83\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"7%\"\u003e\n\u003cp\u003e0.09\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"5%\"\u003e\n\u003cp\u003e0.08\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"6%\"\u003e\n\u003cp\u003e0.003\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"6%\"\u003e\n\u003cp\u003e0.195\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"9%\"\u003e\n\u003cp\u003e0.327\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"16%\"\u003e\n\u003cp\u003e\u003cem\u003eNrf1\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e1.00\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e0.52\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e0.70\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e0.65\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e0.84\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"7%\"\u003e\n\u003cp\u003e0.07\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"5%\"\u003e\n\u003cp\u003e0.05\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"6%\"\u003e\n\u003cp\u003e\u0026lt;0.001\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"6%\"\u003e\n\u003cp\u003e0.016\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"9%\"\u003e\n\u003cp\u003e0.189\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd colspan=\"14\" width=\"100%\"\u003e\n\u003cp\u003eIleal mucosa\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"16%\"\u003e\n\u003cp\u003e\u003cem\u003eAmpk\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e1.00\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e0.96\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e1.09\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e0.87\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e1.40\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"7%\"\u003e\n\u003cp\u003e0.21\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"5%\"\u003e\n\u003cp\u003e0.15\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"6%\"\u003e\n\u003cp\u003e0.295\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"6%\"\u003e\n\u003cp\u003e0.028\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"9%\"\u003e\n\u003cp\u003e0.194\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"16%\"\u003e\n\u003cp\u003e\u003cem\u003eAcc\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e1.00\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e0.93\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e1.18\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e0.76\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e1.32\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"7%\"\u003e\n\u003cp\u003e0.15\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"5%\"\u003e\n\u003cp\u003e0.12\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"6%\"\u003e\n\u003cp\u003e0.476\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"6%\"\u003e\n\u003cp\u003e0.003\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"9%\"\u003e\n\u003cp\u003e0.088\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"16%\"\u003e\n\u003cp\u003e\u003cem\u003eSirt1\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e1.00\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e0.64\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e0.86\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e0.95\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e0.72\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"7%\"\u003e\n\u003cp\u003e0.06\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"5%\"\u003e\n\u003cp\u003e0.05\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"6%\"\u003e\n\u003cp\u003e0.001\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"6%\"\u003e\n\u003cp\u003e0.003\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"9%\"\u003e\n\u003cp\u003e0.081\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"16%\"\u003e\n\u003cp\u003e\u003cem\u003ePgc1\u0026alpha;\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e1.00\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e0.33\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e0.74\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e0.85\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e0.53\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"7%\"\u003e\n\u003cp\u003e0.13\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"5%\"\u003e\n\u003cp\u003e0.11\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"6%\"\u003e\n\u003cp\u003e0.004\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"6%\"\u003e\n\u003cp\u003e0.042\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"9%\"\u003e\n\u003cp\u003e0.067\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"16%\"\u003e\n\u003cp\u003e\u003cem\u003eTfam\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e1.00\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e0.86\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e1.22\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e1.04\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e1.01\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"7%\"\u003e\n\u003cp\u003e0.08\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"5%\"\u003e\n\u003cp\u003e0.07\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"6%\"\u003e\n\u003cp\u003e0.017\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"6%\"\u003e\n\u003cp\u003e0.745\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"9%\"\u003e\n\u003cp\u003e0.854\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"16%\"\u003e\n\u003cp\u003e\u003cem\u003eNrf1\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e0.99\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e1.02\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e1.12\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e1.06\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e1.03\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"7%\"\u003e\n\u003cp\u003e0.09\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"5%\"\u003e\n\u003cp\u003e0.07\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"6%\"\u003e\n\u003cp\u003e0.548\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"6%\"\u003e\n\u003cp\u003e0.763\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"9%\"\u003e\n\u003cp\u003e0.617\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u003csup\u003e1\u003c/sup\u003e Control, basal diets containing 1.59% L-alanine (iso-nitrogen); T1, basal diets containing 1% glutamine, 0.5% glutamate, 0.1% aspartate; T2, energy deficiency diets containing 1% glutamine, 0.5% glutamate, 0.1% aspartate. AMPK: AMP-activated protein kinase, ACC: acetyl-CoA carboxylase, SirT1: Sirtuin 1, PGC1\u003cem\u003e\u0026alpha;\u003c/em\u003e: peroxisome proliferator-activated receptor gamma coactivator 1-\u0026alpha;, Nrf1: nuclear respiratory factor 1, TFAM: mitochondrial transcription factor A.\u003c/p\u003e\n\u003cp\u003e\u003csup\u003e2\u003c/sup\u003e The results were presented as main effects, and the differences among treatments or days were evaluated using Tukey HSD test. \u003csup\u003ea-b \u003c/sup\u003eValues with different letters within the same row are different (\u003cem\u003eP\u003c/em\u003e \u0026lt; 0.05); n = 6.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable S5. Relative protein abundances of AMPK signaling pathway of jejunal and ileal mucosa of weanling piglets \u003csup\u003e1,2\u003c/sup\u003e. \u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"1\" width=\"100%\"\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd rowspan=\"2\" width=\"16%\"\u003e\n\u003cp\u003eItems\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd colspan=\"3\" width=\"25%\"\u003e\n\u003cp\u003eDiet treatments\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd rowspan=\"2\" width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd colspan=\"2\" width=\"16%\"\u003e\n\u003cp\u003eDays post-weaning\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd rowspan=\"2\" width=\"1%\"\u003e\n\u003cp\u003e\u003cem\u003e\u0026nbsp;\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd colspan=\"2\" width=\"13%\"\u003e\n\u003cp\u003e\u003cem\u003eSEM\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd rowspan=\"2\" width=\"1%\"\u003e\n\u003cp\u003e\u003cem\u003e\u0026nbsp;\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd colspan=\"3\" width=\"23%\"\u003e\n\u003cp\u003e\u003cem\u003eP\u003c/em\u003e-value\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003eControl\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003eT 1\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003eT 2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e5\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e21\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"7%\"\u003e\n\u003cp\u003eDiet\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"5%\"\u003e\n\u003cp\u003eDay\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"6%\"\u003e\n\u003cp\u003eDiet\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"6%\"\u003e\n\u003cp\u003eDay\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"9%\"\u003e\n\u003cp\u003eDiet \u0026times; Day\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd colspan=\"14\" width=\"100%\"\u003e\n\u003cp\u003eJejunal mucosa\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"16%\"\u003e\n\u003cp\u003eAMPK\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e0.99\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e1.09\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e1.15\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e1.31\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e0.85\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"7%\"\u003e\n\u003cp\u003e0.05\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"5%\"\u003e\n\u003cp\u003e0.04\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"6%\"\u003e\n\u003cp\u003e0.084\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"6%\"\u003e\n\u003cp\u003e\u0026lt;0.001\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"9%\"\u003e\n\u003cp\u003e\u0026lt;0.001\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"16%\"\u003e\n\u003cp\u003eP-AMPK\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e0.99\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e0.67\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e0.87\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e0.95\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e0.74\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"7%\"\u003e\n\u003cp\u003e0.05\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"5%\"\u003e\n\u003cp\u003e0.04\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"6%\"\u003e\n\u003cp\u003e0.001\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"6%\"\u003e\n\u003cp\u003e0.002\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"9%\"\u003e\n\u003cp\u003e\u0026lt;0.001\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"16%\"\u003e\n\u003cp\u003eACC\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e0.99\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e0.67\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e0.86\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e1.11\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e0.57\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"7%\"\u003e\n\u003cp\u003e0.05\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"5%\"\u003e\n\u003cp\u003e0.04\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"6%\"\u003e\n\u003cp\u003e0.001\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"6%\"\u003e\n\u003cp\u003e\u0026lt;0.001\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"9%\"\u003e\n\u003cp\u003e\u0026lt;0.001\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"16%\"\u003e\n\u003cp\u003eP-ACC\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e0.99\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e0.80\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e1.25\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e1.38\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e0.65\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"7%\"\u003e\n\u003cp\u003e0.09\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"5%\"\u003e\n\u003cp\u003e0.07\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"6%\"\u003e\n\u003cp\u003e0.013\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"6%\"\u003e\n\u003cp\u003e\u0026lt;0.001\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"9%\"\u003e\n\u003cp\u003e0.001\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"16%\"\u003e\n\u003cp\u003eSirT1\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e0.99\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e0.67\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e1.08\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e1.21\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e0.63\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"7%\"\u003e\n\u003cp\u003e0.05\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"5%\"\u003e\n\u003cp\u003e0.04\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"6%\"\u003e\n\u003cp\u003e\u0026lt;0.001\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"6%\"\u003e\n\u003cp\u003e\u0026lt;0.001\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"9%\"\u003e\n\u003cp\u003e\u0026lt;0.001\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"16%\"\u003e\n\u003cp\u003ePGC1a\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e0.99\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e0.88\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e1.12\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e1.36\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e0.64\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"7%\"\u003e\n\u003cp\u003e0.09\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"5%\"\u003e\n\u003cp\u003e0.07\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"6%\"\u003e\n\u003cp\u003e0.172\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"6%\"\u003e\n\u003cp\u003e\u0026lt;0.001\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"9%\"\u003e\n\u003cp\u003e\u0026lt;0.001\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd colspan=\"14\" width=\"100%\"\u003e\n\u003cp\u003eIleal mucosa\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"16%\"\u003e\n\u003cp\u003eAMPK\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e0.99\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e0.81\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e0.55\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e0.99\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e0.58\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"7%\"\u003e\n\u003cp\u003e0.05\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"5%\"\u003e\n\u003cp\u003e0.04\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"6%\"\u003e\n\u003cp\u003e\u0026lt;0.001\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"6%\"\u003e\n\u003cp\u003e\u0026lt;0.001\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"9%\"\u003e\n\u003cp\u003e\u0026lt;0.001\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"16%\"\u003e\n\u003cp\u003eP-AMPK\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e0.99\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e0.48\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e0.90\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e0.88\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e0.70\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"7%\"\u003e\n\u003cp\u003e0.06\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"5%\"\u003e\n\u003cp\u003e0.05\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"6%\"\u003e\n\u003cp\u003e\u0026lt;0.001\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"6%\"\u003e\n\u003cp\u003e0.030\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"9%\"\u003e\n\u003cp\u003e0.040\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"16%\"\u003e\n\u003cp\u003eACC\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e0.99\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e1.34\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e1.19\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e1.25\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd 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width=\"9%\"\u003e\n\u003cp\u003e\u0026lt;0.001\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"16%\"\u003e\n\u003cp\u003ePGC1a\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e0.99\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e0.45\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e1.04\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e1.07\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"8%\"\u003e\n\u003cp\u003e0.59\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"7%\"\u003e\n\u003cp\u003e0.05\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"5%\"\u003e\n\u003cp\u003e0.04\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"1%\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"6%\"\u003e\n\u003cp\u003e\u0026lt;0.001\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"6%\"\u003e\n\u003cp\u003e\u0026lt;0.001\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"9%\"\u003e\n\u003cp\u003e\u0026lt;0.001\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u003csup\u003e1\u003c/sup\u003e Control, basal diets containing 1.59% L-alanine (iso-nitrogen); T1, basal diets containing 1% glutamine, 0.5% glutamate, 0.1% aspartate; T2, energy deficiency diets containing 1% glutamine, 0.5% glutamate, 0.1% aspartate. AMPK: AMP-activated protein kinase, ACC: acetyl-CoA carboxylase, SirT1: Sirtuin 1, PGC1\u003cem\u003e\u0026alpha;\u003c/em\u003e: peroxisome proliferator-activated receptor gamma coactivator 1-\u0026alpha;, Nrf1: nuclear respiratory factor 1, TFAM: mitochondrial transcription factor A.\u003c/p\u003e\n\u003cp\u003e\u003csup\u003e2\u003c/sup\u003e The results were presented as main effects, and the differences among treatments or days were evaluated using Tukey HSD test. \u003csup\u003ea-b \u003c/sup\u003eValues with different letters within the same row are different (\u003cem\u003eP\u003c/em\u003e \u0026lt; 0.05); n = 6.\u003c/p\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"glutamine, glutamate, aspartate, small intestine, energy metabolism ","lastPublishedDoi":"10.21203/rs.3.rs-150010/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-150010/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cstrong\u003eBackground:\u003c/strong\u003e Weaning-stress may cause reduced energy intake for maintenance of mucosal structure. Glutamine (Gln), glutamate (Glu), and aspartate (Asp) are major energy sources for small intestine. This study investigated whether Gln, Glu, and Asp improve the intestinal morphology via regulating the energy metabolism in weaning-piglet. A total of 198 weaned-piglets were assigned to the following treatments: Control (Basal diet + 1.59% L-Alanine); T1 (Basal diet + 1% L-Gln + 0.5% L-Glu + 0.1% L-Asp); T2 (Low energy diet + 1% L-Gln + 0.5% L-Glu + 0.1% L-Asp). Jejunum and ileum were obtained on d-5 or d-21 post-weaning. \u003c/p\u003e\u003cp\u003e\u003cstrong\u003eResults:\u003c/strong\u003e Improved growth performance on d-21 post-weaning were observed in T1 treatment. Both T1 and T2 treatments improved small intestinal morphology by increasing villus height, goblet cell number and decreasing crypt depth on d 5 or d-21 post-weaning. Gln, Glu, and Asp could restore the small intestinal energy homeostasis via replenishing the Krebs’ cycle and down-regulate the AMPK pathway. However, when piglets fed by a low energy diet, Gln, Glu, and Asp are not sufficient to maintain the intestinal energy balance on d-5 post-weaning so that the AMPK, beta-oxidation, and mitochondrial biogenesis are activated to meet the high energy demand of enterocytes. \u003c/p\u003e\u003cp\u003e\u003cstrong\u003eConclusion:\u003c/strong\u003e These data indicated that Gln, Glu, and Asp could restore the energy homeostasis of intestinal mucosa of piglets. And the mucosal energy metabolism showed different responses to the intervention of Gln, Glu, and Asp in piglets with a low energy diet between d5 and d21 post-weaning. These findings provide new information on the nutritional intervention for the insufficient energy intake in weaning-piglet.\u0026nbsp;\u003c/p\u003e","manuscriptTitle":"Glutamine, Glutamate, and Aspartate Improves Morphology and Energy Production of Small Intestine in Piglets with Different Energy Levels Diets","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2021-01-20 18:34:17","doi":"10.21203/rs.3.rs-150010/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"d0127ddc-e7df-4ff5-8cf0-3c1480c020e4","owner":[],"postedDate":"January 20th, 2021","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":1970964,"name":"Animal Science"},{"id":1970965,"name":"Biotechnology and Bioengineering"},{"id":1970966,"name":"Nutrition \u0026 Dietetics"}],"tags":[],"updatedAt":"2021-01-21T16:38:12+00:00","versionOfRecord":[],"versionCreatedAt":"2021-01-20 18:34:17","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-150010","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-150010","identity":"rs-150010","version":["v1"]},"buildId":"rHA-KDH7Qsr4HCuvH75dn","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}