Tagatose Consumption Provokes Metabolic Syndrome Features in Rat Males from Mothers That Consumed Fructose During Their Pregnancy

Tagatose Consumption Provokes Metabolic Syndrome Features in Rat Males from Mothers That Consumed Fructose During Their Pregnancy | 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 Article Tagatose Consumption Provokes Metabolic Syndrome Features in Rat Males from Mothers That Consumed Fructose During Their Pregnancy Elena Fauste, Madelín Pérez-Armas, Cristina Donis, Paola Otero, and 2 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7489857/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 29 Dec, 2025 Read the published version in Molecular Medicine → Version 1 posted 13 You are reading this latest preprint version Abstract Background: Maternal fructose intake induces harmful effects in progeny. However, this sugar is not contraindicated during pregnancy. On the other hand, the use of low-calorie sweeteners, such as tagatose, is increasing. Thus, we have studied whether the consumption of tagatose compared to fructose affects lipid metabolism in the offspring of mothers which were supplemented with fructose during their pregnancy. Methods: Three-month-old male offspring from control or fructose mothers received liquid 10% fructose or tagatose for 21 days. A control group (without any additive) was also included. Biochemical and molecular parameters were determined in plasma, tissues and feces. Results: Both tagatose and fructose consumption caused hypertriglyceridemia in descendants of fructose-fed mothers. Whereas fructose consumption led to a greater hepatic lipogenesis, tagatose supplementation provoked a higher enterohepatic bile acids recirculation, and therefore a higher intestinal lipid absorption and assembly. However, plasma GLP1, a molecule that affects lipid intestinal absorption, was unchanged. Curiously, FGF21, a molecule which regulates lipid and carbohydrate metabolism and is sensitive to GLP1, was augmented in plasma and liver of tagatose-supplemented descendants regardless of their maternal diet. Interestingly, Angiotensin II (Ang II), which can induce FGF21 production, was increased in plasma of all animals supplemented with tagatose. However, the deleterious effects of Ang II were effectively reversed by FGF21 in males from control mothers, but not in descendants of fructose-fed dams. Conclusions: Maternal fructose consumption determines the response of the offspring to tagatose intake, causing an increased intestinal lipid absorption, and metabolic changes that are characteristic of metabolic syndrome such as dyslipidaemia, steatosis and oxidative stress. Fructose pregnancy foetal programming bile acids FGF21 Angiotensin II tagatose Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 1. INTRODUCTION In recent decades, metabolic diseases such as obesity, metabolic syndrome and diabetes have reached epidemic proportions in many countries ( 1 , 2 ). Various studies have shown how metabolic changes that occur during the pre- and postnatal development modulate the risk of developing these diseases, once adult. This phenomenon is called fetal programming and, among all the causal factors, nutrition during gestation is one of the most determinant parameters ( 3 ). Fructose is a monosaccharide found in fruits, honey, sugar beets, and sugar cane. In recent years, there has been an increase in the consumption of added sugars, mainly fructose, in our diet, since they are widely used by the food industry as sweeteners ( 4 ). In fact, fructose is present in a wide variety of processed foods such as industrial pastries, sauces and sugary drinks, which has led to a drastic increase in fructose consumption in the population. Several studies carried out both in experimental animal models and clinical studies in humans have shown that a high fructose intake contributes to the increased incidence of metabolic diseases ( 5 , 6 ). Nevertheless, the consumption of sugary drinks or foods that contain added fructose is not contraindicated during pregnancy. Although in developing countries an increase in the use of added sugars has been found in recent years, in developed countries this trend has changed and the consumption of added sugars has stabilized or even decreased, possibly because social awareness policies are proving effective. Despite these data, the percentage of childhood and adult obesity has continued to increase in recent decades, but more slowly. Thus, in a study conducted by Faruque et al (2019) in the United States, as an example of many other countries, they observed that the drastic increase (annual rate of change = + 1.33) in sugar consumption from the 1970s to the 1990s was followed in parallel by a subsequent exponential growth (+ 0.82 and + 0.97) in the prevalence of obesity that lasted until the 2000s. Interestingly, after the drop (-0.91) in sugar consumption that occurred from the 1990s to the 2010s, a slowdown in the annual increase (+ 0.37) in the prevalence of obesity has been observed from the 2000s onwards( 7 ). In our laboratory, we have developed an "animal model of fetal programming" induced by maternal fructose intake in which the characteristics of metabolic syndrome appear in the offspring naturally or induced after supplementation with liquid fructose. Thus, male offspring from mothers consuming fructose during pregnancy had characteristics of metabolic syndrome, such as hepatic steatosis and hyperinsulinemia; these effects were produced exclusively through epigenetic mechanisms caused by a fetal programming mechanism. Although females from fructose-fed mothers did not appear to be affected by the maternal diet, when they received liquid fructose once adult, they showed an exaggerated response to fructose intake, characterized by hyperlipidaemia and fatty liver ( 8 ). With this and other findings, we could confirm that maternal fructose intake determines the response of the offspring to the diet. These results found in experimental animals could explain why a drastic reduction in the consumption of added sugar has not been accompanied by a parallel decrease in the rate of obesity and various metabolic diseases, as observed by Faruque et al in their study ( 7 ). In fact, it is surprising the case of individuals that despite following a healthy diet, they develop metabolic diseases without any apparent reason. The explanation could be found in epigenetic changes derived from the fact that during pregnancy their mothers consumed large amounts of sugary drinks or processed foods. Related to that, a recently published article would support this hypothesis. Gracner et al studied the exposure to sugars within 1000 days of conception and its impact on diabetes and hypertension. They focused on sugar consumption in the UK population before and after the end of sugar and sweets rationing in 1953. During rationing, sugar intake was at levels within the current recommended dietary guidelines. However, after the end of rationing, consumption almost doubled. Comparing adults conceived pre- or post-rationing, it was observed that the decrease in sugar consumption during the perinatal period reduced the risk of diabetes and hypertension by approximately 35% and 20%, respectively. Interestingly, intrauterine sugar rationing alone accounted for about one-third of the reduction in the risk of developing metabolic disease when adult ( 9 ). Therefore, carbohydrates with sweetening properties and a low caloric value are being investigated to be used as alternative sugars to fructose. Tagatose (an epimer of fructose) is a rare sugar that has antioxidant and prebiotic effects, reduced glycaemic and insulinemic responses, and the potential to improve lipid profile, to induce a lower expression of proinflammatory cytokines, catalase and superoxide dismutase, to decrease lesion area and macrophage infiltration, and to stimulate GLP1 release, therefore constituting an alternative candidate for the treatment of diabetes mellitus and obesity ( 10 , 11 ). In fact, tagatose reached a phase 3 clinical trial to determine its value as an antidiabetic agent. Unlike fructose (4 kcal/g), tagatose is a low-calorie sugar since its energy value is estimated to be 1.5 kcal/g. Currently, tagatose is being added to soft drinks, cereals, chocolate, sweets, caramels, yogurts, ice creams, nutritional supplements and dairy products. Interestingly, only about 20% of consumed tagatose is absorbed by the small intestine, with most of it fermented by colonic bacteria into short-chain fatty acids (SCFA), which are almost completely absorbed. It is mainly metabolized by the liver in a similar way to fructose, with little tagatose reaching the systemic circulation. Tagatose has been recognized by the Food and Drug Administration (FDA) as GRAS ( Generally Recognized As Safe ) and approved as a “new food ingredient” by the European Union, without any restrictions on its use( 12 – 14 ) . With these antecedents and considering that maternal fructose intake determines the response of the offspring to the diet and the increasing use of rare sugars as alternative to the fructose as added sweeteners, we investigated in the present work the effects of maternal fructose consumption on the response of the progeny to tagatose intake for 21 days, in comparison to that of fructose. 2. MATERIAL AND METHODS 2.1. Animals and experimental design An animal model of maternal liquid fructose intake was developed as previously described ( 15 – 17 ). Female Sprague-Dawley rats weighing 200–240 g were fed ad libitum , a standard rat chow diet (Teklad Global 14% Protein Rodent Maintenance Diet, Envigo, USA), and housed under controlled light and temperature conditions (12-h light-dark cycle; 22 ± 1ºC). The experimental protocol was approved by the Ethical Committee for Animal Experimentation of the University San Pablo-CEU and by Autonomous Government of Madrid (ref. numbers 10/206458.9/13 and 10/042445.9/19). Pregnant rats were randomly separated into a control group (no supplementary sugar) and a fructose-supplemented group (fructose 10% wt/vol in drinking water) (7–8 rats per group) throughout gestation ( 17 ). Pregnant rats were allowed to deliver and on the day of birth, each suckling litter was reduced to nine pups per mother. After delivery, both mothers and their pups were maintained with water without any additives and food ad libitum . At 21 days of age, pups were separated by gender and males were fed a standard rat chow diet (Teklad Global 14% Protein Rodent Maintenance Diet, Envigo, USA) and water. The female progeny of each litter was used for a separate experiment. When the male offspring was 3 months old, they were subjected to a new dietary treatment for 21 days regardless of the group of mothers they were born. Male progeny from control or fructose-fed mothers were randomly separated into three experimental groups: control (C, tap water), fructose (F, fructose), and tagatose (T, tagatose), all sugars added as 10% wt/vol in drinking water. Animals within each experimental group were born to different dams to minimize the “litter effect” and the cages contained a maximum of four males to reduce distress. Intake of solid food and liquid per cage were daily recorded and the area under the curve (AUC) for the consumed chow, the ingested liquid and the total amount of ingested calories were calculated. After 21 days of dietary treatment, male offspring were sacrificed. Before this, rats gradually lost consciousness with carbon monoxide. Food and liquid sugar were removed two hours before sacrifice. Blood was collected into EDTA-containing tubes, plasma was obtained by centrifugation and stored at -20ºC until processed. Liver, ileum, heart and lumbar adipose tissue, and the last two feces from the rectum were immediately removed, placed in liquid nitrogen, and kept at -80 ºC until analysis. 2.2. Plasma Determinations Plasma aliquots were used to determine triglycerides and total bile acids (Spinreact, Girona, Spain) using commercial kits. GLP1 (Cusabio, Wuhan, RPC), FGF21 (R&D Systems, USA), copeptin (Cloud-Clone Corp., Wuhan, RPC) and Angiotensin-II (Cusabio, Wuhan, RPC) were determined in plasma samples using specific ELISA kits for rats. 2.3. Tissue Determinations Two hundred milligrams of frozen tissue and one hundred milligrams of feces were immersed in chloroform:methanol 2:1 plus butylhydroxytoluene (BHT) (50 mg/L) and used for lipid extraction following the Folch method ( 27 ). Aliquots of lipid extracts were dried, and the remaining residue was weighed to determine total lipid content. Triglycerides were measured using the procedure described by Carr et al ( 28 ). Briefly, 1 mL of Triton-X 100 1.25% in chloroform was added to 0.3 mL of lipid extracts, dried, and resuspended in 0.5 mL of distilled water. Triglycerides were measured using an enzymatic colorimetric assay (Spinreact, Girona, Spain). Hepatic glycogen was extracted by using ethanol. Briefly, 100 mg of liver was degraded by using 30% KOH and boiling. After that, glycogen was precipitated with ice-cold 99% ethanol for 24h and after centrifugation, the pellet containing glycogen was resuspended in distilled water. Glycogen was hydrolyzed to glucose monomers with an acidic hydrolysis and neutralized before glucose measurement with a colorimetric kit (Spinreact, Gerona, Spain). One hundred milligrams of liver were homogenized in 1,2 mL PBS. After centrifugation, supernatants were used to measure bile acids (Spinreact, Girona, Spain). For bile acid measurement in feces, 0.2 g of feces were dried, and bile acid extraction was performed using 0.5 mL of methanol. After centrifugation, supernatants were used to measure bile acids (Spinreact, Girona, Spain). One hundred milligrams of frozen tissue were homogenized in 0.25 M Tris-HCl, 0.2 M sucrose, and 5 mM dithiothreitol (DTT) buffer at pH 7.4 to determine the oxidative stress state. Thus, the concentration of malondialdehyde (MDA) was measured as a marker of lipid peroxidation using the method previously described( 18 ) (, by measuring the fluorescence of MDA-thiobarbituric acid (TBA) complexes at 515 nm/553 nm excitation/emission wavelengths. Catalase activity was studied by the H 2 O 2 disappearance caused by the activity of this enzyme ( 19 ) This was done by recording the absorbance maximum of H 2 O 2 at 240 nm. Finally, the activity of superoxide dismutase (SOD) was measured using a commercial kit (Merck-Sigma, USA) ). 2.4. RNA extraction and gene expression by qPCR Total RNA was isolated from the tissues using Ribopure (Invitrogen, ThermoFisher Scientific, USA). Total RNA was subjected to DNase I treatment using Turbo DNA-free (Invitrogen, ThermoFisher Scientific, USA), and RNA integrity was confirmed by agarose gel electrophoresis. Afterwards, cDNA was synthesized by oligo(dT)-primed reverse transcription with Superscript II (Invitrogen, ThermoFisher Scientific, USA). qPCRs were performed using a CXF96® Touch (Bio-Rad, California, USA). The reaction solution was carried out in a volume of 20 µl, containing 10 pmol of both forward and reverse primers, 10x SYBR Premix Ex Taq (Takara Bio Inc., Japan), and the appropriate nanograms of the cDNA stock solution. Rps29 was used as a reference gene for qPCR. The primer sequences were designed using primer-BLAST software (NCBI) ( 20 ). Samples were analysed in duplicate on each assay. Amplification of non-specific targets was discarded using the melting curve analysis method for each amplicon. qPCR efficiency and linearity were assessed by optimization of the standard curves for each target. The transcription was quantified with CFX Maestro 2.0 software (Bio-Rad, California, USA) using the efficiency correction method ( 21 ). 2.5. Statistical Analysis Results were expressed as means ± S.E. Treatment effects were analyzed by two-way analysis of variance (ANOVA) with maternal diet (M) and progeny diet (D) as factors. Then, the Bonferroni test was used for post hoc analysis to identify the source of significant variance. Data that were not normally distributed were log-transformed to achieve data normality. Significant differences (p < 0.05) were indicated either with asterisks (*) between groups of animals receiving different nutritional treatments but belonging to the same dietary group of mothers or hash symbols (#) between groups of rats receiving the same treatment but coming from different dietary groups of mothers. The eta2 (η2) parameter (i.e., the proportion of the total variance that was attributed to the corresponding effect (M, D, or the M × D interaction)) was also provided (Table S1 ). All statistical analysis were performed using SPSS version 29 computer program. 3. RESULTS 3.1. Food, liquid, and total caloric intake were not modified by tagatose consumption. The sweetening power of tagatose is only slightly less than that of sucrose with a relative sweetness of 92% when compared in 10% solutions, and it is not so high as that of fructose, which is the sweetest of all naturally occurring carbohydrates( 22 ) The relative sweetness of fructose has been reported in the range of 1.2–1.8 times that of sucrose. Curiously, whereas the intake of liquid was significantly increased in males consuming 10% fructose when compared to males that received water without additives, as previously published ( 23 ), the AUC of ingested liquid trended to be lower in males receiving 10% tagatose versus their respective control males, although without reaching statistical significance in any case (Fig. 1 A). These effects were observed regardless of their maternal diet. Male offspring from control mothers consumed 47% of their total calories from fructose, while in males from fructose-fed mothers, around 41% of the total amount of energy was acquired from fructose. On the other hand, tagatose is considered to provide 1.5 Kcal/g, due to its low absorption rate, compared to sucrose or fructose that provide 4.0 Kcal/g ( 24 ), consequently, only 5% and 5.5% of total calories come from tagatose in the progeny from control and fructose-fed mothers, respectively. According to this, in both groups subjected to liquid fructose, solid diet consumption was similarly reduced to compensate for the calories obtained from fructose. However, in the case of tagatose, this compensatory effect for the calories ingested from sugar was not observed in descendants from control mothers but it was found in progeny from fructose-fed dams, although without being significantly different (Fig. 1 B). Thus, in males from control mothers, the total amount of ingested energy was the same in descendants drinking water without additives as in males supplemented with tagatose, and slightly lower in males consuming tagatose versus those consuming only water in progeny from fructose-fed mothers (Fig. 1 C). As previously observed by us ( 8 ), this compensatory effect turned out to be inefficient in the case of descendants consuming fructose. Interestingly, these differences in the total calorie intake observed between the experimental groups, mainly in comparison to the animals that consumed fructose, were not reflected in the final body weight since there were no differences between the different dietetic experimental groups, neither in progeny from control mothers nor in descendants from fructose-fed dams (Table S2 ). And the same situation was found for the weights of diverse organs: liver, heart and lumbar adipose tissue. The only noteworthy differences were found between progeny from control and fructose-fed mothers (indicated by hash symbols), but these modifications are due to the significantly lower body weight observed in all descendants of fructose-fed mothers before starting the dietary treatments, as already we have described in previous studies ( 8 , 25 ). 3.2. Tagatose induced ChREBP transcriptional activity in ileum but not in liver Given the different intake of tagatose versus fructose observed in all descendants, we were first interested in confirming whether these sugars were producing any effect on their own metabolism. It is known that ChREBP (carbohydrate responsive element binding protein) is a transcription factor that responds to sugar consumption ( 26 ). Moreover, ChREBP activity in vivo appears to be more responsive to sugars other than glucose and, in fact, it is potently activated by fructose ingestion. ChREBP co-ordinately regulates the expression of all three fructolytic enzymes: ketohexokinase (KHK), aldolase b (ALDOB), and triokinase and FMN cyclase (TKFC). Interestingly, the metabolism of tagatose is identical to that of fructose ( 10 ) Therefore, the expression of these enzymes and the specific transporter for the entry of these sugars (glucose transporter 5, GLUT5) to the cells was determined both in the intestine (the first organ able to metabolize them) and the liver (which is supposed to be the main organ in charge of metabolizing them). Curiously, the effects observed were again quite different between the two carbohydrates. Unexpectedly, tagatose was metabolized more efficiently than fructose in the ileum, even though it had been ingested in significantly smaller amounts. Thus, tagatose (but not fructose) induced significantly GLUT5 gene expression in comparison to the other dietary treatments in all descendants, being more evident in descendants from control mothers (Fig. 2 A). Moreover, tagatose (in contrast to fructose) induced the gene expression of the three tagatolytic enzymes in all descendants, although, in this case, the effect was more pronounced and significant in progeny from fructose-fed mothers (Figs. 2 B- 2 D). Regarding the liver, the situation was different. Both sugar transport and its metabolism were preferentially induced by fructose and barely activated by tagatose. In addition, tagatose seemed to be more efficient in affecting its metabolism in the progeny of control mothers than in the progeny of fructose-fed dams (Figs. 2 E- 2 H). Thus, fructose activated GLUT5, KHK, ALDOB, and TKFC hepatic gene expression showing a similar profile than the intake of liquid fructose (Fig. 1 A), that is, in a more evident way in descendants from control dams than in descendants from fructose-fed mothers (Fig. 2 E- 2 H). Tagatose, however, induced GLUT5 and TFKC, but not KHK and ALDOB, being this effect only found in males from control mothers. Interestingly, tagatolysis was less induced in the intestine of these animals (Fig. 2 A- 2 D) than in progeny from fructose-fed dams and, therefore, some amount of tagatose is supposed to be able to reach the liver, leading to the effects observed in this tissue (Fig. 2 E- 2 H). 3.3. Tagatose induced dyslipidemia in the progeny of fructose-fed dams due to an increased intestinal reabsorption and recirculation of bile acids As described in the previous section, tagatose and fructose were absorbed, reaching the systemic circulation and consequently metabolized by the liver and/or intestine. Since tagatose had been ingested in a significant smaller amount than fructose by all descendants, regardless of maternal diet, it was expected that progeny fed tagatose would not show dyslipidaemia. However, surprisingly, this was not the case. Triglyceridemia was significantly modified after the 21 days of nutritional treatment with both fructose and tagatose and, more importantly, this effect was uniquely observed in progeny from fructose-fed dams (Fig. 3 A). It is widely described that fructose is able to raise plasma triglycerides ( 27 ), however, tagatose is considered to improve plasma lipid profile ( 12 ). These results suggest a worsened lipid profile in descendants of fructose-fed mothers consuming liquid fructose or tagatose. To elucidate the underlying mechanisms of this sugar-induced hypertriglyceridemia, and considering the well-established role of fructose in stimulating lipogenesis and inhibiting β-oxidation, we assessed the hepatic expression of key genes involved in fatty acid metabolism, specifically the lipogenic gene stearoyl-CoA desaturase-1 (SCD1) and the catabolic gene carnitine palmitoyl transferase 1 (CPT1) ( 27 ). Thus, fructose intake produced an overexpression of SCD1 compared to control groups, no matter the mothers' diet, while tagatose consumption did not modify SCD1 gene expression (Fig. 3 B). This increment in fatty acid synthesis caused by fructose could be providing more substrate for triglycerides production and their exportation to blood, leading to an increase in triglyceridemia. Thus, hepatic gene expression of microsomal triglyceride transfer protein (MTTP) (Fig. 3 C), a protein involved in very low-density lipoprotein (VLDL) assembly, showed a non-significant trend to increase in males consuming fructose, regardless of the diet of their mothers. Related to fatty acid catabolism, CPT1 gene expression, the enzyme that controls fatty acid entry to mitochondria for its oxidation, tended to be reduced in males from control mothers that were supplemented with fructose, whereas no changes were observed in the corresponding males from fructose-fed mothers (Fig. 3 D). However, tagatose consumption did not provoke any changes in CPT1 expression in males from control dams, but it produced a non-significant increase in descendants from fructose-fed mothers (Fig. 3 D). Since tagatose intake did not appear to affect lipogenesis, secretion of triglycerides-rich lipoproteins into the bloodstream or fatty acid oxidation (mechanisms that could account for the tagatose-induced hypertriglyceridemia observed in the offspring of fructose-fed mothers), we evaluated alternative pathways. One of these pathways that could affect the lipid profile in blood is the intestinal absorption of lipids. As shown in Fig. 3 E, fecal triglycerides remained unchanged among the three groups of descendants from control mothers. In contrast, in the progeny of fructose-fed dams, sugars intake led to a diminution in fecal triglyceride content, which reached statistical significance in the tagatose group when compared to control animals. Consistent with these findings, ileal gene expression of MTTP, the enzyme responsible for assembly in chylomicrons of absorbed lipids, presented no differences in the offspring of control mothers but, importantly, a significant increase was observed in tagatose-supplemented descendants from fructose-fed mothers compared to the control group (Fig. 3 F). Related to that, bile acids are molecules secreted into bile to facilitate intestinal absorption of fats ( 28 ). In the present study, fecal bile acids did not change in descendants of control mothers after consuming fructose or tagatose compared to the control group (Fig. 4 A). Interestingly, however, progeny from fructose-fed mothers that received liquid fructose or tagatose did present a non-significant increment in fecal bile acids, being this trend more pronounced in the tagatose group (Fig. 4 A). Curiously, this same profile was seen in both plasma (Fig. 4 B) bile acids and hepatic (Fig. 4 C) bile acids. Thus, in the offspring of fructose-fed mothers that consumed tagatose, plasma bile acids levels were significantly augmented compared to the corresponding control group (Fig. 4 B) and hepatic bile acids content (Fig. 4 C) was significantly increased in comparison to both control and fructose groups. These results may indicate an alteration in the enterohepatic bile acid recirculation along the intestine-plasma-liver axis. A higher recirculation of bile acids would lead to an increased intestinal content of bile acids which would facilitate lipids emulsification and absorption. This mechanism could contribute to the lipid alterations observed in the offspring of fructose-fed mothers that consumed tagatose. In accordance with this, all genes involved in bile acid reabsorption in the ileum were overexpressed in the progeny from fructose-fed mothers that consumed tagatose (Fig. 4 D-G). Thus, whereas in the progeny from control mothers no differences were observed among the three experimental groups, in descendants from fructose-fed mothers, tagatose administration did significantly increase the gene expression of ASBT (apical sodium-bile acid transporter) (Fig. 4 D) and OST (organic solute transporter), both type alpha (OSTa) (Fig. 4 F) and type beta (OSTb) (Fig. 4 G) compared to the control group. In the case of IBABP (ileal bile acid-binding protein) gene expression, the significant increase in the tagatose group was found in comparison to males that consumed fructose (Fig. 4 E). Moreover, these increases turned out to be also significant for ASBT (Fig. 4 D) and OSTb (Fig. 4 G) when comparing males that consumed tagatose from fructose-fed mothers versus descendants of control mothers. Consequently, tagatose intake in descendants from fructose-supplemented mothers would be provoking a higher bile acid entry to the ileum (ASBT), an enhanced bile acid circulation across the enterocyte (IBABP), and a more pronounced bile acid export to blood (OST). These results would highlight how foetal programming induced by maternal fructose intake can modulate the response to a nutrient in progeny. 3.4. Tagatose modified plasma FGF21 but not GLP1 levels in the progeny independently of the mother´s diet. Intestinal production of triglyceride-rich lipoproteins has also been shown to be affected by glucagon-like peptide 1 (GLP-1) ( 29 ), an incretin produced by the gut that has been reported to play a key role in inducing satiety, reducing food intake and controlling obesity. That is why the study of this molecule is attracting increasing interest among the healthcare scientific community. Interestingly, tagatose has been shown to stimulate the release of GLP-1 ( 12 ), although short-chain fatty acids (SCFA) produced by bacterial fermentation of poorly absorbed sugars into the gut may also stimulate GLP-1 ( 10 ). In fact, GLP-1 is released in response to the presence of diverse nutrients in the intestine. Expression level of the ileal proglucagon gene (a precursor of GLP-1) has been shown to be increased in the presence of glucose, free fatty acids and SCFA in the distal gut ( 30 ) In the present study, tagatose intake led to an increased ileal gene expression of transporters/receptors of nutrients such as GLUT5 (Fig. 2 A) for sugars, ABST (Fig. 4 D) for bile acids, G protein-coupled receptors (GPR41 and GPR43) for SCFAs, and Takeda G protein-coupled receptor (TGR5) for bile acids (Table 1 ), being the effect more evident in the progeny of fructose-fed mothers. Interestingly, the upregulation of these genes may help to explain why proglucagon gene expression was induced in descendants of fructose-fed dams consuming tagatose (Fig. 5 A), reaching statistical significance in comparison to the control group. However, this increase in proglucagon gene expression in males from fructose-fed mothers consuming tagatose was not accompanied by an augmented expression of PC1/3 (proprotein convertase 1/3) (Fig. 5 B), the enzyme responsible for processing proglucagon to the active form of GLP1, nor by a decreased expression of Dpp4 (dipeptidyl peptidase 4), the enzyme that degrades GLP-1 (Fig. 5 C). Consequently, plasma GLP1 levels did not differ among all experimental groups (Fig. 5 D). It is important to note that we could not measure active GLP-1 levels in serum and the data shown in Fig. 5 D reflect total GLP-1 levels. Table 1 Ileal and hepatic gene (mRNA) expression of control (C), fructose- (F), and tagatose-supplemented (T) male progeny from control or fructose-fed mothers. CONTROL MOTHERS FRUCTOSE MOTHERS CONTROL FRUCTOSE TAGATOSE p CONTROL FRUCTOSE TAGATOSE p Ileal mRNA Gene Expression (a.u) GPR41 1.033±0.104 1.213±0.128 1.264±0.116 0.761±0.076 0.718±0.111# 1.255±0.192 * (FC vs FT) * (FF vs FT) GPR43 1.057±0.129 1.468±0.1778 1.637±0.205 * (CC vs CT) 1.069±0.131 1.039±0.203 1.448±0.132 TGR5 1.132±0.087 1.78±0.178 1.687±0.263 * (CC vs CF) 1.082±0.063 1.313±0.138 1.548±0.076 ** (FC vs FT) Hepatic mRNA Gene Expression (a.u) MCT1 1.002±0.077 1.048±0.104 0.977±0.043 1.158±0.112 1.044±0.056 1.335±0.127## HDAC1 1.024±0.08 1.003±0.136 1.290±0.105 0.975±0.047 0.868±0.121 1.062±0.095 HDAC3 0.943±0.057 0.990±0.092 1.034±0.096 1.055±0.086 1.042±0.048 0.882±0.04 PDK4 1.080±0.162 1.003±0.010 1.583±0.196 1.067±0.109 0.993±0.178 1.483±0.086 CIDEC 1,008±0,048 1.105±0.108 1.360±0.089 * (CC vs CT) 1.106±0.076 1.100±0.123 1.374±0.087 VLDLR 1.037±0.096 0.883±0.042 1.033±0.113 1.679±0.257 2.035±0.388## 1.945±0.271## CD36 0.0014±0.0002 0.0016±0.0001 0.0024±0.0002 0.0022±0.0006 0.0017±0.0004 0.0011±0.0002# Ileal and hepatic levels of specific mRNA genes are shown. Ileum mRNA expression represents GLP1 signalling pathway genes, and liver mRNA expression represents both SCFA and PPAR alpha signalling pathway genes. Relative target gene mRNA levels were measured by Real Time PCR as explained in Materials and Methods, normalized to Rps29 levels and expressed in arbitrary units (a.u.). Data are means ± S.E. from 7 to 8 litters. Asterisks denote a significant difference (*, p < 0.05; **, p < 0.01; ***, p < 0.001) between the groups with a different diet but the same mothers ́ diet. Hash symbols denote a significant difference (#, p < 0.05; ##, p < 0.01; ###, p < 0.001) as compared to the control mothers (groups with the same diet but different mothers ́ diet). fructose. The first letter indicates whether the mothers had been supplied with tap water during pregnancy (C: control) or liquid fructose (F); and the second letter indicates the nutritional treatment without (C: control) or with additives, fructose (F) or tagatose (T), when they were adults. GLP1: glucagon-like protein 1; and GLP1 signalling: GPR: G protein-coupled receptors; TGR: Takeda G protein-coupled receptor. SCFA: short-chain fatty acids; and SCFA signalling: MCT1: monocarboxylate transporter 1 protein; HDAC: Histone deacetylases; PPAR: peroxisome proliferator-activated receptor; and PPAR signalling: PDK: pyruvate dehydrogenase kinase; CIDEC: cell death-inducing DNA fragmentation factor-like effector C; VLDLR: very low-density lipoprotein receptor; CD36: cluster of differentiation 36. On the other hand, it has been reported that FGF21, a hormone that also responds to nutrients, can regulate sugar preference and, in addition, its production is induced by GLP-1 analogues ( 31 ). In the present work we found that FGF21 plasma levels were augmented in sugar-supplemented descendants, regardless of the diet consumed by their mothers (Fig. 5 E). Curiously, plasma FGF21 levels paralleled the different amount of liquid fructose ingested by descendants, which was influenced by their mothers´diet (Fig. 1 A). Importantly, tagatose-fed descendants, who had consumed approximately 10 times less amount of sugar than the fructose-fed groups, showed elevated plasma FGF21 levels. This increase in the tagatose-supplemented animals was significant compared to the control group in the progeny of control mothers and compared to the control and fructose-fed groups in descendants from fructose-fed dams (Fig. 5 E). Considering that FGF21 is mainly synthesized in the liver and that our results indicate that tagatose, unlike fructose, barely reaches this organ, it was very striking to find that tagatose intake induced hepatic FGF21 gene expression whereas fructose did not. Moreover, this tagatose-induced activation was more pronounced in the offspring of fructose-fed dams than in those of control mothers (Fig. 5 F). 3.5. Tagatose intake increased plasma levels of Ang II which produced worse effects in the offspring of fructose-fed mothers. It has been shown that, in response to carbohydrate intake, the liver produces FGF21 which acts on the hypothalamus to selectively suppress sugar intake. Interestingly, in the present study, after tagatose consumption, plasma FGF21 levels and sugar intake were inversely related, that is, tagatose increased plasma FGF21, and this reduced sugar appetite (Fig. 5 E versus Fig. 1 A). In contrast, in fructose-fed animals we found a direct relationship between sugar intake and FGF21 levels. Carbohydrates are known to activate hepatic ChREBP, a transcription factor that promotes FGF21 production in the liver ( 32 ). However, as previously mentioned, tagatose consumption hardly affected the hepatic expression of some ChREBP target genes such as KHK and ALDOB (Figs. 2 F and 2 G). These data suggest that ChREBP is likely not involved in the tagatose-induced activation of hepatic FGF21 production. Tagatose is minimally absorbed in the intestine and, thus, it has been proposed as a prebiotic agent capable of selectively stimulating the growth of specific gut microbiota and affecting host health ( 33 ). By doing this, tagatose induces microbiota that produce beneficial compounds such as butyrate. This SCFA has been demonstrated to stimulate hepatic FGF21 gene expression by inhibiting histone deacetylase 3 (HDAC3) which suppresses the activity of peroxisome proliferator-activated receptor type alpha (PPARα) ( 34 ). And, to note, FGF21 is a well-known PPAR alpha target gene in liver ( 35 ). However, none of the nutritional interventions here used altered the gene expression of monocarboxylate transporter 1 (MCT1), which permits SCFA entry to the cell, or that of HDAC1 or HDAC3 (Table 1 ). Thus, exploring the role of PPARα in the tagatose-induced activation of FGF21, we found that whereas CPT1 (Fig. 3 D), PDK4 and CIDEC gene expression (Table 1 ) observed in the tagatose groups could reflect an activation of PPARα, the findings observed for SCD1 (Fig. 3 B), VLDLR and CD36 gene expression did not show changes in that sense (Table 1 ). Moreover, it has been suggested that part of the effect of PPARα on hepatic ketogenesis may be mediated by induction of the PPARα target FGF21; however, plasma ketone bodies trended to be diminished in all descendants consuming carbohydrates regardless of their mothers´diet [166.1 ± 28.4; 92.9 ± 19.3; and 95.6 ± 5.1 for control, fructose- and tagatose-fed descendants from control mothers; 206.3 ± 41.2; 143.1 ± 41.1; and 165.7 ± 52.3 µM for control, fructose- and tagatose-fed males from fructose-fed dams]. Importantly, angiotensin II (Ang II), a molecule that promotes inflammation, oxidative stress, vascular injury, fatty liver and insulin resistance ( 36 ), has been shown to be able to increase serum FGF21 levels and hepatic FGF21 gene expression, possibly as a compensatory and protective response against these harmful effects induced by Ang II ( 37 , 38 ). As shown in Fig. 6 A, and in accordance with the results found in hepatic FGF21 gene expression (Fig. 5 G), tagatose intake produced an increase in plasma Ang II in both descendants from control mothers and males from fructose-fed dams. This augmentation observed in tagatose-fed progeny turned out to be significant versus fructose-fed animals in descendants of control mothers and versus the control group in males from fructose-fed mothers. In accordance, the angiotensin-converting enzyme (ACE), that converts AngI to AngII, although its hepatic gene expression was not affected by any dietary treatment (Fig. 6 B), ileal ACE gene expression was significantly augmented after carbohydrate ingestion in males from control mothers and sharply increased after tagatose intake in progeny from fructose-fed dams. In fact, these tagatose-mediated effects were significant when compared to the other two groups of descendants from fructose-fed dams. Moreover, tagatose-supplemented males from fructose-fed mothers showed a significant higher expression of ileal ACE than the descendants of control mothers fed with the same sugar (denoted by a hash symbol) suggesting a fetal programming mediated effect (Fig. 6 C). Additionally, Ang II is well-known to stimulate vasopressin secretion ( 39 ), a molecule that is related to liquid ingestion, carbohydrate (fructose, glucose or HFCS) intake and it has been proposed as a key mediator in the fructose-induced metabolic syndrome ( 40 ). However, we measured plasma copeptin, an analogue and stable peptide derived from the precursor of vasopressin, and no differences were found among all the experimental groups [46.1 ± 6.8; 36.8 ± 5.4; and 49.9 ± 4.9 for control, fructose- and tagatose-fed descendants from control mothers; 49.4 ± 6.9; 46.0 ± 0.8; and 39.9 ± 3.0 pg/mL for control, fructose- and tagatose-fed males from fructose-fed dams]. Nevertheless, despite this lack of effect mediated by carbohydrate intake in copeptin levels, the mRNA gene expression of vasopressin 1a receptor (AVP1AR) showed a reduction in the liver of fructose-fed animals from control mothers in consonance to previous studies ( 40 ) and a significant reduction in tagatose-supplemented males from these same mothers (Fig. 6 D). Curiously, in the progeny from fructose-fed mothers, the effect produced by fructose ingestion was not observed whereas tagatose intake significantly diminished AVP1AR expression versus both control and fructose groups. Furthermore, this fetal programming effect was more evident in heart AVP1AR expression (Fig. 6 E), since no changes were observed among the three experimental groups of descendants from control mothers, but a significant reduction was found in tagatose-fed males from fructose-fed mothers in comparison to the other two groups and also when compared to the corresponding tagatose-fed group from control mothers (hash symbol). Interestingly, Andres-Hernando et al observed in AVP1AR-KO mice worse metabolic features of metabolic syndrome than in the wild-type group ( 40 ). In view of these interesting results observed in tagatose-fed males, that is, elevated levels of plasma FGF21 trying to compensate the high levels of plasma Ang II and a lower presence of the vasopressin 1a receptor, we decided to evaluate oxidative stress and lipid and glucose dysfunction parameters in both liver and heart. Curiously, FGF21 appeared to counteract the effects of Ang II in the liver but not in the heart. This protective effect was more evident in the progeny from control mothers than in those from fructose-fed dams. Thus, in the liver, none of the nutritional interventions altered MDA levels, catalase activity or superoxide dismutase activity, regardless of the maternal diet (Table 2 ). In contrast, in the heart, there was a tendency for these oxidative markers to be modified by carbohydrate intake in progeny from control mothers, although statistically significant differences were only found in tagatose-fed descendants from fructose-fed mothers. Specifically, catalase activity was significantly decreased compared to the fructose group, while SOD activity was elevated versus both fructose and control groups (Table 2 ). As shown in Fig. 7 A, whereas liver glycogen content remained unchanged in all experimental groups, cardiac glycogen content (Fig. 7 B) was augmented by carbohydrate intake in the progeny from control mothers, becoming significantly different in tagatose-fed males versus the control group. This tagatose-mediated increase in cardiac glycogen was even more pronounced and significant in descendants from fructose-fed dams versus the other two groups. Additionally, a similar profile to the one observed in oxidative parameters and glycogen was also found in the triglycerides content. Thus, hepatic steatosis was not observed in any case, although a slight non-significant lipid accretion was found in tagatose-fed animals from fructose-fed dams (Fig. 7 C). Interestingly, whereas cardiac triglycerides content did not change among the three experimental groups in males from control dams, a significant triglycerides deposit was found in males from fructose-fed dams after consuming tagatose (Fig. 7 D) compared to the control group. Table 2 Hepatic and cardiac oxidative stress parameters of control (C), fructose- (F), and tagatose-supplemented (T) male progeny from control or fructose-fed mothers. Data are means ± S.E. from 7 to 8 litters. Asterisks denote a significant difference (*, p < 0.05; **, p < 0.01; ***, p < 0.001) between the groups with a different diet but the same mothers ́ diet. Hash symbols denote a significant difference (#, p < 0.05; ##, p < 0.01; ###, p < 0.001) as compared to the control mothers (groups with the same diet but different mothers ́ diet). fructose. The first letter indicates whether the mothers had been supplied with tap water during pregnancy (C: control) or liquid fructose (F); and the second letter indicates the nutritional treatment without (C: control) or with additives, fructose (F) or tagatose (T), when they were adults. MDA: malondialdehyde; Cat: catalase activity; SOD: superoxide dismutase activity. CONTROL MOTHERS FRUCTOSE MOTHERS CONTROL FRUCTOSE TAGATOSE p CONTROL FRUCTOSE TAGATOSE p Liver oxidative-stress parameters MDA (mmol/g tissue) 15.552±1.357 15.398±1.737 17.383±1.616 14.553±0.463 14.447±0.664 18.227±1.924 Cat (mU/mg prot) 1391.6±120.8 1036.4±103.7 1373.4±117.8 1578.1±121.5 1682.3±190.1## 1663.9±232.6 SOD (U/mg prot) 43.33±2.67 42.37±1.43 44.35±1.67 40.79±0.47 43.11±0.33 44.65±2.32 Heart oxidative-stress parameters MDA (nmol/g tissue) 36.17±4.21 52.33±7.34 57.87±12.77 34.38±4.53 48.70±12.83 38.55±5.64 Cat (mU/mg prot) 42.91±2.65 39.52±2.42 36.07±2.20 40.67±1.08 43.48±1.90 33.82±1.54 ** (FF vs FT) SOD (U/mg prot) 23.54±0.52 24.57±0.49 27.04±1.38 23.78±0.92 23.42±1.51 28.83±1.21 * (FC vs FT) ** (FF vs FT) Therefore, this oxidative stress and accumulation of fuel stores suggest that tagatose consumption induces cardiac metabolic dysregulation in the offspring of fructose-fed mothers ( 41 , 42 ) that ultimately would lead to cardiac dysfunction. Moreover, we have previously described a similar scenario in which FGF21 was able to protect against lipid accretion and oxidative stress influenced by maternal nutrition and in a tissue-dependent manner ( 15 ). 4. DISCUSSION The urgent need to reduce the high prevalence of metabolic diseases, especially those driven by modifiable lifestyle factors such as "diseases related to processed food", makes dietary changes essential. One key aspect of these changes involves reducing added sugars intake, particularly fructose or its derivatives, and replacing them with alternative sugars or sweeteners, such as rare sugars. This shift has already been taking place since the first decades of this century and, fortunately, it has contributed to a gradual slowdown in the development of metabolic diseases. However, the decrease in added sugar consumption has not been paralleled by a proportional reduction in the incidence of metabolic diseases such as obesity ( 7 ). We believe this discrepancy is because high fructose consumption was common among pregnant women prior to these dietary changes. Consequently, it is logical that the offspring of these mothers continue to exhibit a high incidence of metabolic diseases and that their response to diet is affected by a fetal programming mechanism due to the maternal intake of added sugar ( 9 ). Given the increasing incorporation of rare sugars to replace fructose, sucrose or HFCS in our diet, we decided to find out the impact of maternal fructose intake on offspring´s metabolic response to consumption of the rare sugar tagatose. One of the most striking results found is that, despite the low intake of tagatose (about 5% of total caloric intake), it was able to stimulate the gene expression of enzymes and transporters of the tagatolysis pathway in the intestine, in a more pronounced way in descendants of fructose-fed mothers, and, interestingly, with almost no influence on hepatic tagatolysis. In contrast, fructose, which accounted for about 45% of total caloric intake, had the opposite effect, that is, it did not modify intestinal fructolysis and clearly affected the gene expression of liver fructolysis enzymes and transporters ( 43 ). These data suggest that the effects observed mainly in the offspring of fructose-fed mothers are more related to a metabolite produced by intestinal tagatolysis than to tagatose itself. Another relevant finding was that tagatose intake affected triglyceridemia in a way that was clearly influenced by fetal programming, since it induced hypertriglyceridemia in the offspring of fructose-fed mothers, but not in those of control mothers. Fructose intake also elevated triglyceridemia in that same group of descendants. Interestingly, while fructose-mediated effect was mainly caused by a higher expression of lipogenic genes in the liver, the effect of tagatose appeared to be associated with a higher gene expression of bile acid transporters in the intestine. This result suggests a greater enterohepatic recirculation of bile acids, which led to a greater lipid absorption and packaging of in the intestine. Such a mechanism would explain the hyperlipidemia observed in these descendants and aligns with findings previously described by us and others although using another type of diet ( 23 , 44 ). Interestingly, gene expression of most transporters and receptors of nutrients, such as sugars, short-chain fatty acids, and bile acids, was increased in the ileum after tagatose intake, particularly in the offspring of fructose-fed dams. These transporters and receptors have been directly related to intestinal production of proglucagon in response to these nutrients ( 30 ). In fact, consistent with this, we observed that the tagatose-induced upregulation of the expression of nutrient transporters and receptors was accompanied by an increase in the expression of proglucagon. However, plasma GLP1 levels did not reflect this observed rise in its precursor molecule. In addition, GLP2, which is also produced from proglucagon and whose high levels would be more in line with the greater lipid absorption and packaging found in the descendants of fructose-fed mothers consuming tagatose, was not affected by the different treatments ( 29 ). Unfortunately, total GLP was measured and not the active form that would be more informative. One of the most striking results was to find that both tagatose intake and, as expected, fructose consumption increased plasma levels of FGF21 ( 32 ). Interestingly, although tagatose (as mentioned above) seemed to barely reach the liver as it failed to stimulate tagatolysis in this organ, it was able to clearly increase the hepatic expression of FGF21. FGF21 has been described as a molecule capable of decreasing the sweet taste preference ( 32 ). Therefore, it was paradoxical to find that, whereas fructose intake was directly related to plasma FGF21 levels, the relationship with tagatose consumption was clearly opposite and more in line with what has been previously described, that is, the tagatose-induced increase in FGF21 could be the cause of the low intake of this sugar observed in the offspring. We were therefore interested in discovering the mechanism by which tagatose intake increases the hepatic expression of FGF21. We found that classic effectors such as ChREBP, PPAR alpha and even the mediation of SCFA that could be produced in the intestine by unabsorbed tagatose, failed to explain it adequately ( 14 ). However, a less studied regulatory molecule in the metabolism of FGF21, such as angiotensin II, was found to be increased in plasma after tagatose intake. This increase was consistent with the findings observed in hepatic expression of FGF21, so we could affirm that tagatose consumption increased Ang II levels and, with it, hepatic production of FGF21. Interestingly, Ang II has also been shown to be involved in the production of vasopressin ( 39 ), a molecule that could influence the lower fluid intake seen in animals that consumed tagatose, as has been described in the case of fructose ( 40 ). However, levels of copeptin (a stable form of vasopressin) were not changed by any of the treatments. On the other hand, the hepatic expression of its receptor 1a (AVP1AR) was modified by fructose intake, which decreased it, confirming previous findings by other authors ( 40 ). Notably, tagatose reduced the hepatic gene expression of this receptor more drastically than fructose in all descendants and, furthermore, it also diminished AVP1AR gene expression in the heart, although in this case this effect was only observed in descendants of fructose-mothers. Considering that the lack of this receptor has been related to the appearance of more severe symptoms of metabolic syndrome, this interesting result warrants further investigation in future studies ( 40 ). It has been described that Ang II promotes the production of FGF21 to counteract its adverse effects ( 37 , 38 ). Thus, in the present study, FGF21 manages to counteract the negative effects of Ang II in the liver of all descendants fed tagatose. However, this was not the case in the heart of males from fructose-fed mothers since oxidative stress and a clear accumulation of glycogen and triglycerides were observed after the intake of tagatose. These dysfunctions could be an initial biomarker of cardiac metabolic dysregulation, a very common situation observed in diabetes or metabolic syndrome ( 41 ). 5. CONCLUSIONS Therefore, the consumption of alternative sugars to fructose and/or HFCS, such as tagatose, by descendants of mothers who consumed fructose during their pregnancy could lead to adverse effects more likely than in descendants of control mothers. The current study highlights a novel interaction between tagatose consumption and angiotensin II signaling, leading to increased hepatic FGF21 expression even in the absence of direct hepatic sugar metabolism. This unique metabolic response raises concerns regarding its systemic effects, particularly in individuals with prenatal exposure to excess fructose. The inability of FGF21 to mitigate Ang II-associated dysfunction in cardiac tissue suggests early biomarkers of metabolic disruption. These data call for a re-evaluation of the metabolic safety of rare sugars like tagatose. Overall, these findings emphasize the key role of maternal nutrition during gestation, particularly the quality and quantity of sugar intake, as it can seriously affect the long-term metabolic health of the progeny through fetal programming mechanisms. Consequently, these observations along with previous reports strongly support the adherence to the WHO guidelines which recommend limiting the intake of simple sugars in processed foods and sugary drinks to less than 10% of total daily caloric intake. Importantly, as evidenced by the present study, such limitations should also include rare sugars such as tagatose. Declarations Acknowledgements The authors thank Jose M. Garrido and his team for their help in handling the rats. Author contributions C.B. and E.F. conceived and designed the study. E.F., M.P-A., C.D., P.O. and M.I.P. contributed reagents/materials/analysis tools for gene expression studies and parameter analysis. E.F., P.O. and M.I.P. handled the animals. M.I.P. analysed the data. C.B. and E.F. wrote the paper. All authors had access to the study data and had reviewed and approved the final manuscript. Funding This work was supported by funds from the Ministerio de Ciencia e Innovación (MCIN): PID2020-118054RB-I00/AEI/10.13039/501100011033 and PID2023-152756OB-I00/AEI/10.13039/501100011033. Elena Fauste was previously supported, and Madelín Pérez-Armas is supported with FPU fellowships from MCIN. Availability of data and materials All data generated or analysed during this study are included in this published article [and its supplementary information files]. Ethics approval and consent to participate The experimental protocol was approved by the Ethical Committee for Animal Experimentation of the University San Pablo-CEU and by Autonomous Government of Madrid (ref. numbers 10/206458.9/13 and 10/042445.9/19). Consent for publication Not applicable. Competing interests There are no conflicts of interest to declare. References Carrera-Bastos P, Fontes-Villalba M, O’Keefe JH, Lindeberg S, Cordain L. The western diet and lifestyle and diseases of civilization. Res Rep Clin Cardiol. 2011;2(1):15–35. Kopp W. How Western Diet And Lifestyle Drive The Pandemic Of Obesity And Civilization Diseases. Diabetes Metab Syndr Obes. 2019;12:2221–36. Howie GJ, Sloboda DM, Kamal T, Vickers MH. Maternal nutritional history predicts obesity in adult offspring independent of postnatal diet. J Physiol. 2009;587(Pt 4):905–15. White JS, Nicklas TA. High-fructose corn syrup use in beverages: composition, manufacturing, properties, consumption, and health effects. 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Supplementary Files FaustePerezArmasDonisetalSupplTable1.docx FaustePerezArmasDonisetalSupplTable2.docx Cite Share Download PDF Status: Published Journal Publication published 29 Dec, 2025 Read the published version in Molecular Medicine → Version 1 posted Editorial decision: Revision requested 07 Oct, 2025 Reviews received at journal 07 Oct, 2025 Reviews received at journal 04 Oct, 2025 Reviews received at journal 29 Sep, 2025 Reviews received at journal 23 Sep, 2025 Reviewers agreed at journal 23 Sep, 2025 Reviewers agreed at journal 21 Sep, 2025 Reviewers agreed at journal 20 Sep, 2025 Reviewers agreed at journal 18 Sep, 2025 Reviewers invited by journal 18 Sep, 2025 Editor assigned by journal 01 Sep, 2025 Submission checks completed at journal 01 Sep, 2025 First submitted to journal 29 Aug, 2025 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. 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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-7489857","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":521767554,"identity":"d5d52c52-a477-491d-a70a-216f20b7eff4","order_by":0,"name":"Elena Fauste","email":"","orcid":"","institution":"San Pablo CEU University","correspondingAuthor":false,"prefix":"","firstName":"Elena","middleName":"","lastName":"Fauste","suffix":""},{"id":521767555,"identity":"8116ba96-b1c6-4ed8-b9dd-8d586e2909af","order_by":1,"name":"Madelín Pérez-Armas","email":"","orcid":"","institution":"San Pablo CEU University","correspondingAuthor":false,"prefix":"","firstName":"Madelín","middleName":"","lastName":"Pérez-Armas","suffix":""},{"id":521767556,"identity":"b40c5de0-7929-4b99-9627-9a411f266c51","order_by":2,"name":"Cristina Donis","email":"","orcid":"","institution":"San Pablo CEU University","correspondingAuthor":false,"prefix":"","firstName":"Cristina","middleName":"","lastName":"Donis","suffix":""},{"id":521767557,"identity":"1d63b84a-7bb0-4ea4-8856-3c1948585eb7","order_by":3,"name":"Paola Otero","email":"","orcid":"","institution":"San Pablo CEU University","correspondingAuthor":false,"prefix":"","firstName":"Paola","middleName":"","lastName":"Otero","suffix":""},{"id":521767558,"identity":"2354901b-fb77-4273-84d5-c404e2f2b0c1","order_by":4,"name":"Mª Isabel Panadero","email":"","orcid":"","institution":"San Pablo CEU 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16:45:33","extension":"xml","order_by":14,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":155535,"visible":true,"origin":"","legend":"","description":"","filename":"c47370a879154ce6bfc293ae1625a3601structuring.xml","url":"https://assets-eu.researchsquare.com/files/rs-7489857/v1/771dfda45849e610e7e78529.xml"},{"id":92434641,"identity":"4b8a179f-2f5e-4299-9311-b355219838e8","added_by":"auto","created_at":"2025-09-29 16:45:33","extension":"html","order_by":15,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":167401,"visible":true,"origin":"","legend":"","description":"","filename":"earlyproof.html","url":"https://assets-eu.researchsquare.com/files/rs-7489857/v1/914332f8c254a65e6afec682.html"},{"id":92434636,"identity":"19f11af5-f00d-403f-9f57-96a6413b6cb7","added_by":"auto","created_at":"2025-09-29 16:45:32","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":60248,"visible":true,"origin":"","legend":"\u003cp\u003eFood, liquid, and total caloric intake were not modified by tagatose consumption. (A) AUC ingested liquid, (B) AUC consumed chow, and (C) total amount of ingested energy from control (C, empty bar), fructose- (F, light grey bar), and tagatose-supplemented (T, dark grey bar) male progeny from control (left panel) or fructose-fed (right panel) mothers. Data are means ± S.E. from 7-8 litters. Asterisks denote a significant difference (*, p \u0026lt; 0.05; **, p \u0026lt; 0.01; ***, p \u0026lt; 0.001) between the groups under the crossbar (groups with a different diet but the same mother´s diet). Hash symbols denote a significant difference (#, p \u0026lt; 0.05; ##, p \u0026lt; 0.01; ###, p \u0026lt; 0.001) as compared to the control mothers (groups with the same diet but different mother´s diet). AUC: area under the curve.\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-7489857/v1/355c2ec0214362bf27c61eba.png"},{"id":92435131,"identity":"dddea89f-701b-461e-b196-f06e56f8f8e5","added_by":"auto","created_at":"2025-09-29 16:53:32","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":99564,"visible":true,"origin":"","legend":"\u003cp\u003eTagatose induced ChREBP transcriptional activity in ileum but not in liver. (A) GLUT5, (B) KHK, (C) Aldo B, (D) TFKC mRNA ileal gene expression, and (E) GLUT5, (F) KHK, (G) Aldo B, (H) TFKC mRNA liver gene expression from control (C, empty bar), fructose- (F, light grey bar), and tagatose-supplemented (T, dark grey bar) young male progeny from control (left panel) or fructose-fed (right panel) mothers. Data are means ± S.E. from 7-8 litters. Relative target gene mRNA levels were measured by Real-Time PCR as explained in Materials and Methods, normalized to Rps29 levels and expressed in arbitrary units (a.u.). Asterisks denote a significant difference (*, p \u0026lt; 0.05; **, p \u0026lt; 0.01; ***, p \u0026lt; 0.001) between the groups under the crossbar (groups with a different diet but the same mother´s diet). Hash symbols denote a significant difference (#, p \u0026lt; 0.05; ##, p \u0026lt; 0.01; ###, p \u0026lt; 0.001) as compared to the control mothers (groups with the same diet but different mother´s diet). ChREBP: carbohydrate-responsive element-binding protein; GLUT: glucose transporter; KHK: ketohexokinase; Aldo: aldolase; TKFC: triokinase and FMN Cyclase.\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-7489857/v1/2361859706567db345267c74.png"},{"id":92433551,"identity":"b2176557-52da-4c93-a648-9e3a5acbe1b6","added_by":"auto","created_at":"2025-09-29 16:37:32","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":96690,"visible":true,"origin":"","legend":"\u003cp\u003eTagatose produced dyslipidemia in the progeny of fructose-fed dams. (A) ratio day 21/day 0 plasma triglycerides, (B) SCD1, (C) MTTP, and (D) CPT1 mRNA liver gene expression; and (E) fecal triglycerides, and (F) MTTP mRNA ileal gene expression from control (C, empty bar), fructose- (F, light grey bar), and tagatose-supplemented (T, dark grey bar) male progeny from control (left panel) or fructose-fed (right panel) mothers. Data are means ± S.E. from 7-8 litters. Relative target gene mRNA levels were measured by Real-Time PCR as explained in Materials and Methods, normalized to Rps29 levels and expressed in arbitrary units (a.u.). Asterisks denote a significant difference (*, p \u0026lt; 0.05; **, p \u0026lt; 0.01; ***, p \u0026lt; 0.001) between the groups under the crossbar (groups with a different diet but the same mother´s diet). Hash symbols denote a significant difference (#, p \u0026lt; 0.05; ##, p \u0026lt; 0.01; ###, p \u0026lt; 0.001) as compared to the control mothers (groups with the same diet but different mother´s diet). SCD1: stearoyl-CoA desaturase-1; MTTP: microsomal triglyceride transfer protein; CPT1: carnitine palmitoyl transferase 1.\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-7489857/v1/e9767ed56de6686e87710094.png"},{"id":92434634,"identity":"09a2ec70-e058-4789-88e0-0f54623a8a78","added_by":"auto","created_at":"2025-09-29 16:45:32","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":94485,"visible":true,"origin":"","legend":"\u003cp\u003eTagatose produced an increased intestinal reabsorption and recirculation of bile acids in the progeny of fructose-fed dams. (A) Bile acids levels in feces, (B) plasma, (C) and the liver, and (D) ASBT, (E) IBABP, (F) OSTa, and (G) OSTb mRNA ileal gene expression from control (C, empty bar), fructose- (F, light grey bar), and tagatose-supplemented (T, dark grey bar) male progeny from control (left panel) or fructose-fed (right panel) mothers. Data are means ± S.E. from 7-8 litters. Relative target gene mRNA levels were measured by Real-Time PCR as explained in Materials and Methods, normalized to Rps29 levels and expressed in arbitrary units (a.u.). Asterisks denote a significant difference (*, p \u0026lt; 0.05; **, p \u0026lt; 0.01; ***, p \u0026lt; 0.001) between the groups under the crossbar (groups with a different diet but the same mother´s diet). Hash symbols denote a significant difference (#, p \u0026lt; 0.05; ##, p \u0026lt; 0.01; ###, p \u0026lt; 0.001) as compared to the control mothers (groups with the same diet but different mother´s diet). ASBT: apical sodium-bile acid transporter; IBABP: ileal bile acid-binding protein; OST: organic solute transporter.\u003c/p\u003e","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-7489857/v1/fb3c70111e0e08bf59d56001.png"},{"id":92433556,"identity":"5e08a74a-4497-4c4a-92c0-6ec95932babb","added_by":"auto","created_at":"2025-09-29 16:37:32","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":89860,"visible":true,"origin":"","legend":"\u003cp\u003eTagatose modified plasma FGF21 but not GLP1 levels in the progeny independently of the mother´s diet. (A) Proglucagon, (B) PC1/3 and (C) DPP4 mRNA ileal gene expression, and (D) plasma GLP1 and (E) FGF21 levels, and (G) FGF21 mRNA hepatic gene expression from control (C, empty bar), fructose- (F, light grey bar), and tagatose-supplemented (T, dark grey bar) male progeny from control (left panel) or fructose-fed (right panel) mothers. Data are means ± S.E. from 7-8 litters. Relative target gene mRNA levels were measured by Real-Time PCR as explained in Materials and Methods, normalized to Rps29 levels and expressed in arbitrary units (a.u.). Asterisks denote a significant difference (*, p \u0026lt; 0.05; **, p \u0026lt; 0.01; ***, p \u0026lt; 0.001) between the groups under the crossbar (groups with a different diet but the same mother´s diet). Hash symbols denote a significant difference (#, p \u0026lt; 0.05; ##, p \u0026lt; 0.01; ###, p \u0026lt; 0.001) as compared to the control mothers (groups with the same diet but different mother´s diet). PC1/3: proprotein convertase 1/3; DPP4: dipeptidyl peptidase 4; GLP1: glucagon-like protein 1; FGF21: fibroblast growth factor 21.\u003c/p\u003e","description":"","filename":"5.png","url":"https://assets-eu.researchsquare.com/files/rs-7489857/v1/4fef193331e93c04d0bf9559.png"},{"id":92435132,"identity":"a8a95bf5-71c7-46dc-af3c-6fa49530e693","added_by":"auto","created_at":"2025-09-29 16:53:32","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":65432,"visible":true,"origin":"","legend":"\u003cp\u003eTagatose modified plasma Ang II levels in the progeny regardless of the diet of the mother. (A) Plasma angiotensin II; (B) hepatic ACE and (C) ileal ACE mRNA gene expression; and (D) hepatic AVPR1A and (E) cardiac AVPR1A mRNA gene expression from control (C, empty bar), fructose- (F, light grey bar), and tagatose-supplemented (T, dark grey bar) male progeny from control (left panel) or fructose-fed (right panel) mothers. Data are means ± S.E. from 7-8 litters. Relative target gene mRNA levels were measured by Real-Time PCR as explained in Materials and Methods, normalized to Rps29 levels and expressed in arbitrary units (a.u.). Asterisks denote a significant difference (*, p \u0026lt; 0.05; **, p \u0026lt; 0.01; ***, p \u0026lt; 0.001) between the groups under the crossbar (groups with a different diet but the same mother´s diet). Hash symbols denote a significant difference (#, p \u0026lt; 0.05; ##, p \u0026lt; 0.01; ###, p \u0026lt; 0.001) as compared to the control mothers (groups with the same diet but different mother´s diet). Ang II: angiotensin II; ACE: angiotensin I converting enzyme; AVPR1A: arginine vasopressin receptor 1A gene.\u003c/p\u003e","description":"","filename":"6.png","url":"https://assets-eu.researchsquare.com/files/rs-7489857/v1/4d469a0d8ac85d8746ad4f71.png"},{"id":92433559,"identity":"133233fe-9e29-4edf-8fdd-05ecc31893d5","added_by":"auto","created_at":"2025-09-29 16:37:32","extension":"png","order_by":7,"title":"Figure 7","display":"","copyAsset":false,"role":"figure","size":68096,"visible":true,"origin":"","legend":"\u003cp\u003eTagatose produced cardiac accretion of glycogen and triglycerides mainly in fructose-fed mother descendants. (A) hepatic and (B) cardiac glycogen contents, and (C) hepatic and (D) cardiac triglycerides from control (C, empty bar), fructose- (F, light grey bar), and tagatose-supplemented (T, dark grey bar) male progeny from control (left panel) or fructose-fed (right panel) mothers. Data are means ± S.E. from 7-8 litters. Relative target gene mRNA levels were measured by Real-Time PCR as explained in Materials and Methods, normalized to Rps29 levels and expressed in arbitrary units (a.u.). Asterisks denote a significant difference (*, p \u0026lt; 0.05; **, p \u0026lt; 0.01; ***, p \u0026lt; 0.001) between the groups under the crossbar (groups with a different diet but the same mother´s diet). Hash symbols denote a significant difference (#, p \u0026lt; 0.05; ##, p \u0026lt; 0.01; ###, p \u0026lt; 0.001) as compared to the control mothers (groups with the same diet but different mother´s diet).\u003c/p\u003e","description":"","filename":"7.png","url":"https://assets-eu.researchsquare.com/files/rs-7489857/v1/937323e5b4a3f2be4300e1bc.png"},{"id":99545392,"identity":"7b5e4c0b-2d8a-4659-ac29-4125a09138bd","added_by":"auto","created_at":"2026-01-05 16:07:00","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1515310,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-7489857/v1/b7244854-2be3-4858-91e6-6ee6b917d177.pdf"},{"id":92433563,"identity":"4bc8ae0c-3a79-4f44-bd9f-b7a6091c0fbf","added_by":"auto","created_at":"2025-09-29 16:37:33","extension":"docx","order_by":2,"title":"","display":"","copyAsset":false,"role":"supplement","size":44757,"visible":true,"origin":"","legend":"","description":"","filename":"FaustePerezArmasDonisetalSupplTable1.docx","url":"https://assets-eu.researchsquare.com/files/rs-7489857/v1/ac0ebece21d92827ec793df0.docx"},{"id":92433558,"identity":"3f47cca3-b6af-4216-8fdc-a51e76a19fdb","added_by":"auto","created_at":"2025-09-29 16:37:32","extension":"docx","order_by":3,"title":"","display":"","copyAsset":false,"role":"supplement","size":27239,"visible":true,"origin":"","legend":"","description":"","filename":"FaustePerezArmasDonisetalSupplTable2.docx","url":"https://assets-eu.researchsquare.com/files/rs-7489857/v1/685274ef948b164bb832c695.docx"}],"financialInterests":"No competing interests reported.","formattedTitle":"\u003cp\u003eTagatose Consumption Provokes Metabolic Syndrome Features in Rat Males from Mothers That Consumed Fructose During Their Pregnancy\u003c/p\u003e","fulltext":[{"header":"1. INTRODUCTION","content":"\u003cp\u003eIn recent decades, metabolic diseases such as obesity, metabolic syndrome and diabetes have reached epidemic proportions in many countries (\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e, \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e). Various studies have shown how metabolic changes that occur during the pre- and postnatal development modulate the risk of developing these diseases, once adult. This phenomenon is called fetal programming and, among all the causal factors, nutrition during gestation is one of the most determinant parameters (\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eFructose is a monosaccharide found in fruits, honey, sugar beets, and sugar cane. In recent years, there has been an increase in the consumption of added sugars, mainly fructose, in our diet, since they are widely used by the food industry as sweeteners (\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e). In fact, fructose is present in a wide variety of processed foods such as industrial pastries, sauces and sugary drinks, which has led to a drastic increase in fructose consumption in the population. Several studies carried out both in experimental animal models and clinical studies in humans have shown that a high fructose intake contributes to the increased incidence of metabolic diseases (\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e, \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e). Nevertheless, the consumption of sugary drinks or foods that contain added fructose is not contraindicated during pregnancy.\u003c/p\u003e\u003cp\u003eAlthough in developing countries an increase in the use of added sugars has been found in recent years, in developed countries this trend has changed and the consumption of added sugars has stabilized or even decreased, possibly because social awareness policies are proving effective. Despite these data, the percentage of childhood and adult obesity has continued to increase in recent decades, but more slowly. Thus, in a study conducted by Faruque et al (2019) in the United States, as an example of many other countries, they observed that the drastic increase (annual rate of change\u0026thinsp;=\u0026thinsp;+\u0026thinsp;1.33) in sugar consumption from the 1970s to the 1990s was followed in parallel by a subsequent exponential growth (+\u0026thinsp;0.82 and +\u0026thinsp;0.97) in the prevalence of obesity that lasted until the 2000s. Interestingly, after the drop (-0.91) in sugar consumption that occurred from the 1990s to the 2010s, a slowdown in the annual increase (+\u0026thinsp;0.37) in the prevalence of obesity has been observed from the 2000s onwards(\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eIn our laboratory, we have developed an \"animal model of fetal programming\" induced by maternal fructose intake in which the characteristics of metabolic syndrome appear in the offspring naturally or induced after supplementation with liquid fructose. Thus, male offspring from mothers consuming fructose during pregnancy had characteristics of metabolic syndrome, such as hepatic steatosis and hyperinsulinemia; these effects were produced exclusively through epigenetic mechanisms caused by a fetal programming mechanism. Although females from fructose-fed mothers did not appear to be affected by the maternal diet, when they received liquid fructose once adult, they showed an exaggerated response to fructose intake, characterized by hyperlipidaemia and fatty liver (\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e). With this and other findings, we could confirm that maternal fructose intake determines the response of the offspring to the diet.\u003c/p\u003e\u003cp\u003eThese results found in experimental animals could explain why a drastic reduction in the consumption of added sugar has not been accompanied by a parallel decrease in the rate of obesity and various metabolic diseases, as observed by Faruque et al in their study (\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e). In fact, it is surprising the case of individuals that despite following a healthy diet, they develop metabolic diseases without any apparent reason. The explanation could be found in epigenetic changes derived from the fact that during pregnancy their mothers consumed large amounts of sugary drinks or processed foods. Related to that, a recently published article would support this hypothesis. Gracner et al studied the exposure to sugars within 1000 days of conception and its impact on diabetes and hypertension. They focused on sugar consumption in the UK population before and after the end of sugar and sweets rationing in 1953. During rationing, sugar intake was at levels within the current recommended dietary guidelines. However, after the end of rationing, consumption almost doubled. Comparing adults conceived pre- or post-rationing, it was observed that the decrease in sugar consumption during the perinatal period reduced the risk of diabetes and hypertension by approximately 35% and 20%, respectively. Interestingly, intrauterine sugar rationing alone accounted for about one-third of the reduction in the risk of developing metabolic disease when adult (\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eTherefore, carbohydrates with sweetening properties and a low caloric value are being investigated to be used as alternative sugars to fructose. Tagatose (an epimer of fructose) is a rare sugar that has antioxidant and prebiotic effects, reduced glycaemic and insulinemic responses, and the potential to improve lipid profile, to induce a lower expression of proinflammatory cytokines, catalase and superoxide dismutase, to decrease lesion area and macrophage infiltration, and to stimulate GLP1 release, therefore constituting an alternative candidate for the treatment of diabetes mellitus and obesity (\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e, \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e). In fact, tagatose reached a phase 3 clinical trial to determine its value as an antidiabetic agent.\u003c/p\u003e\u003cp\u003eUnlike fructose (4 kcal/g), tagatose is a low-calorie sugar since its energy value is estimated to be 1.5 kcal/g. Currently, tagatose is being added to soft drinks, cereals, chocolate, sweets, caramels, yogurts, ice creams, nutritional supplements and dairy products. Interestingly, only about 20% of consumed tagatose is absorbed by the small intestine, with most of it fermented by colonic bacteria into short-chain fatty acids (SCFA), which are almost completely absorbed. It is mainly metabolized by the liver in a similar way to fructose, with little tagatose reaching the systemic circulation. Tagatose has been recognized by the Food and Drug Administration (FDA) as GRAS (\u003cem\u003eGenerally Recognized As Safe\u003c/em\u003e) and approved as a \u0026ldquo;new food ingredient\u0026rdquo; by the European Union, without any restrictions on its use(\u003cspan additionalcitationids=\"CR13\" citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e) .\u003c/p\u003e\u003cp\u003eWith these antecedents and considering that maternal fructose intake determines the response of the offspring to the diet and the increasing use of rare sugars as alternative to the fructose as added sweeteners, we investigated in the present work the effects of maternal fructose consumption on the response of the progeny to tagatose intake for 21 days, in comparison to that of fructose.\u003c/p\u003e"},{"header":"2. MATERIAL AND METHODS","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e\u003ch2\u003e2.1. Animals and experimental design\u003c/h2\u003e\u003cp\u003eAn animal model of maternal liquid fructose intake was developed as previously described (\u003cspan additionalcitationids=\"CR16\" citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e). Female Sprague-Dawley rats weighing 200\u0026ndash;240 g were fed \u003cem\u003ead libitum\u003c/em\u003e, a standard rat chow diet (Teklad Global 14% Protein Rodent Maintenance Diet, Envigo, USA), and housed under controlled light and temperature conditions (12-h light-dark cycle; 22\u0026thinsp;\u0026plusmn;\u0026thinsp;1\u0026ordm;C). The experimental protocol was approved by the Ethical Committee for Animal Experimentation of the University San Pablo-CEU and by Autonomous Government of Madrid (ref. numbers 10/206458.9/13 and 10/042445.9/19).\u003c/p\u003e\u003cp\u003ePregnant rats were randomly separated into a control group (no supplementary sugar) and a fructose-supplemented group (fructose 10% wt/vol in drinking water) (7\u0026ndash;8 rats per group) throughout gestation (\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e). Pregnant rats were allowed to deliver and on the day of birth, each suckling litter was reduced to nine pups per mother. After delivery, both mothers and their pups were maintained with water without any additives and food \u003cem\u003ead libitum\u003c/em\u003e. At 21 days of age, pups were separated by gender and males were fed a standard rat chow diet (Teklad Global 14% Protein Rodent Maintenance Diet, Envigo, USA) and water. The female progeny of each litter was used for a separate experiment. When the male offspring was 3 months old, they were subjected to a new dietary treatment for 21 days regardless of the group of mothers they were born. Male progeny from control or fructose-fed mothers were randomly separated into three experimental groups: control (C, tap water), fructose (F, fructose), and tagatose (T, tagatose), all sugars added as 10% wt/vol in drinking water. Animals within each experimental group were born to different dams to minimize the \u0026ldquo;litter effect\u0026rdquo; and the cages contained a maximum of four males to reduce distress. Intake of solid food and liquid per cage were daily recorded and the area under the curve (AUC) for the consumed chow, the ingested liquid and the total amount of ingested calories were calculated. After 21 days of dietary treatment, male offspring were sacrificed. Before this, rats gradually lost consciousness with carbon monoxide. Food and liquid sugar were removed two hours before sacrifice. Blood was collected into EDTA-containing tubes, plasma was obtained by centrifugation and stored at -20\u0026ordm;C until processed. Liver, ileum, heart and lumbar adipose tissue, and the last two feces from the rectum were immediately removed, placed in liquid nitrogen, and kept at -80 \u0026ordm;C until analysis.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec4\" class=\"Section2\"\u003e\u003ch2\u003e2.2. Plasma Determinations\u003c/h2\u003e\u003cp\u003ePlasma aliquots were used to determine triglycerides and total bile acids (Spinreact, Girona, Spain) using commercial kits. GLP1 (Cusabio, Wuhan, RPC), FGF21 (R\u0026amp;D Systems, USA), copeptin (Cloud-Clone Corp., Wuhan, RPC) and Angiotensin-II (Cusabio, Wuhan, RPC) were determined in plasma samples using specific ELISA kits for rats.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec5\" class=\"Section2\"\u003e\u003ch2\u003e2.3. Tissue Determinations\u003c/h2\u003e\u003cp\u003eTwo hundred milligrams of frozen tissue and one hundred milligrams of feces were immersed in chloroform:methanol 2:1 plus butylhydroxytoluene (BHT) (50 mg/L) and used for lipid extraction following the Folch method (\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e). Aliquots of lipid extracts were dried, and the remaining residue was weighed to determine total lipid content. Triglycerides were measured using the procedure described by Carr et al (\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e). Briefly, 1 mL of Triton-X 100 1.25% in chloroform was added to 0.3 mL of lipid extracts, dried, and resuspended in 0.5 mL of distilled water. Triglycerides were measured using an enzymatic colorimetric assay (Spinreact, Girona, Spain).\u003c/p\u003e\u003cp\u003eHepatic glycogen was extracted by using ethanol. Briefly, 100 mg of liver was degraded by using 30% KOH and boiling. After that, glycogen was precipitated with ice-cold 99% ethanol for 24h and after centrifugation, the pellet containing glycogen was resuspended in distilled water. Glycogen was hydrolyzed to glucose monomers with an acidic hydrolysis and neutralized before glucose measurement with a colorimetric kit (Spinreact, Gerona, Spain).\u003c/p\u003e\u003cp\u003eOne hundred milligrams of liver were homogenized in 1,2 mL PBS. After centrifugation, supernatants were used to measure bile acids (Spinreact, Girona, Spain). For bile acid measurement in feces, 0.2 g of feces were dried, and bile acid extraction was performed using 0.5 mL of methanol. After centrifugation, supernatants were used to measure bile acids (Spinreact, Girona, Spain).\u003c/p\u003e\u003cp\u003eOne hundred milligrams of frozen tissue were homogenized in 0.25 M Tris-HCl, 0.2 M sucrose, and 5 mM dithiothreitol (DTT) buffer at pH 7.4 to determine the oxidative stress state. Thus, the concentration of malondialdehyde (MDA) was measured as a marker of lipid peroxidation using the method previously described(\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e) (, by measuring the fluorescence of MDA-thiobarbituric acid (TBA) complexes at 515 nm/553 nm excitation/emission wavelengths. Catalase activity was studied by the H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e disappearance caused by the activity of this enzyme (\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e) This was done by recording the absorbance maximum of H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e at 240 nm. Finally, the activity of superoxide dismutase (SOD) was measured using a commercial kit (Merck-Sigma, USA) ).\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec6\" class=\"Section2\"\u003e\u003ch2\u003e2.4. RNA extraction and gene expression by qPCR\u003c/h2\u003e\u003cp\u003eTotal RNA was isolated from the tissues using Ribopure (Invitrogen, ThermoFisher Scientific, USA). Total RNA was subjected to DNase I treatment using Turbo DNA-free (Invitrogen, ThermoFisher Scientific, USA), and RNA integrity was confirmed by agarose gel electrophoresis. Afterwards, cDNA was synthesized by oligo(dT)-primed reverse transcription with Superscript II (Invitrogen, ThermoFisher Scientific, USA). qPCRs were performed using a CXF96\u0026reg; Touch (Bio-Rad, California, USA). The reaction solution was carried out in a volume of 20 \u0026micro;l, containing 10 pmol of both forward and reverse primers, 10x SYBR Premix Ex Taq (Takara Bio Inc., Japan), and the appropriate nanograms of the cDNA stock solution. Rps29 was used as a reference gene for qPCR. The primer sequences were designed using primer-BLAST software (NCBI) (\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eSamples were analysed in duplicate on each assay. Amplification of non-specific targets was discarded using the melting curve analysis method for each amplicon. qPCR efficiency and linearity were assessed by optimization of the standard curves for each target. The transcription was quantified with CFX Maestro 2.0 software (Bio-Rad, California, USA) using the efficiency correction method (\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e).\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec7\" class=\"Section2\"\u003e\u003ch2\u003e2.5. Statistical Analysis\u003c/h2\u003e\u003cp\u003eResults were expressed as means\u0026thinsp;\u0026plusmn;\u0026thinsp;S.E. Treatment effects were analyzed by two-way analysis of variance (ANOVA) with maternal diet (M) and progeny diet (D) as factors. Then, the Bonferroni test was used for \u003cem\u003epost hoc\u003c/em\u003e analysis to identify the source of significant variance. Data that were not normally distributed were log-transformed to achieve data normality. Significant differences (p\u0026thinsp;\u0026lt;\u0026thinsp;0.05) were indicated either with asterisks (*) between groups of animals receiving different nutritional treatments but belonging to the same dietary group of mothers or hash symbols (#) between groups of rats receiving the same treatment but coming from different dietary groups of mothers. The eta2 (η2) parameter (i.e., the proportion of the total variance that was attributed to the corresponding effect (M, D, or the M \u0026times; D interaction)) was also provided (Table \u003cspan refid=\"MOESM1\" class=\"InternalRef\"\u003eS1\u003c/span\u003e). All statistical analysis were performed using SPSS version 29 computer program.\u003c/p\u003e\u003c/div\u003e"},{"header":"3. RESULTS","content":"\u003cdiv id=\"Sec9\" class=\"Section2\"\u003e\u003ch2\u003e3.1. Food, liquid, and total caloric intake were not modified by tagatose consumption.\u003c/h2\u003e\u003cp\u003eThe sweetening power of tagatose is only slightly less than that of sucrose with a relative sweetness of 92% when compared in 10% solutions, and it is not so high as that of fructose, which is the sweetest of all naturally occurring carbohydrates(\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e) The relative sweetness of fructose has been reported in the range of 1.2\u0026ndash;1.8 times that of sucrose. Curiously, whereas the intake of liquid was significantly increased in males consuming 10% fructose when compared to males that received water without additives, as previously published (\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e), the AUC of ingested liquid trended to be lower in males receiving 10% tagatose versus their respective control males, although without reaching statistical significance in any case (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eA). These effects were observed regardless of their maternal diet.\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003eMale offspring from control mothers consumed 47% of their total calories from fructose, while in males from fructose-fed mothers, around 41% of the total amount of energy was acquired from fructose. On the other hand, tagatose is considered to provide 1.5 Kcal/g, due to its low absorption rate, compared to sucrose or fructose that provide 4.0 Kcal/g (\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e), consequently, only 5% and 5.5% of total calories come from tagatose in the progeny from control and fructose-fed mothers, respectively. According to this, in both groups subjected to liquid fructose, solid diet consumption was similarly reduced to compensate for the calories obtained from fructose. However, in the case of tagatose, this compensatory effect for the calories ingested from sugar was not observed in descendants from control mothers but it was found in progeny from fructose-fed dams, although without being significantly different (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eB). Thus, in males from control mothers, the total amount of ingested energy was the same in descendants drinking water without additives as in males supplemented with tagatose, and slightly lower in males consuming tagatose versus those consuming only water in progeny from fructose-fed mothers (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eC). As previously observed by us (\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e), this compensatory effect turned out to be inefficient in the case of descendants consuming fructose.\u003c/p\u003e\u003cp\u003eInterestingly, these differences in the total calorie intake observed between the experimental groups, mainly in comparison to the animals that consumed fructose, were not reflected in the final body weight since there were no differences between the different dietetic experimental groups, neither in progeny from control mothers nor in descendants from fructose-fed dams (Table \u003cspan refid=\"MOESM2\" class=\"InternalRef\"\u003eS2\u003c/span\u003e). And the same situation was found for the weights of diverse organs: liver, heart and lumbar adipose tissue. The only noteworthy differences were found between progeny from control and fructose-fed mothers (indicated by hash symbols), but these modifications are due to the significantly lower body weight observed in all descendants of fructose-fed mothers before starting the dietary treatments, as already we have described in previous studies (\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e, \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e).\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec10\" class=\"Section2\"\u003e\u003ch2\u003e3.2. Tagatose induced ChREBP transcriptional activity in ileum but not in liver\u003c/h2\u003e\u003cp\u003e\u003cdiv class=\"BlockQuote\"\u003e\u003cp\u003eGiven the different intake of tagatose versus fructose observed in all descendants, we were first interested in confirming whether these sugars were producing any effect on their own metabolism. It is known that ChREBP (carbohydrate responsive element binding protein) is a transcription factor that responds to sugar consumption (\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e). Moreover, ChREBP activity in vivo appears to be more responsive to sugars other than glucose and, in fact, it is potently activated by fructose ingestion. ChREBP co-ordinately regulates the expression of all three fructolytic enzymes: ketohexokinase (KHK), aldolase b (ALDOB), and triokinase and FMN cyclase (TKFC). Interestingly, the metabolism of tagatose is identical to that of fructose (\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e) Therefore, the expression of these enzymes and the specific transporter for the entry of these sugars (glucose transporter 5, GLUT5) to the cells was determined both in the intestine (the first organ able to metabolize them) and the liver (which is supposed to be the main organ in charge of metabolizing them). Curiously, the effects observed were again quite different between the two carbohydrates. Unexpectedly, tagatose was metabolized more efficiently than fructose in the ileum, even though it had been ingested in significantly smaller amounts. Thus, tagatose (but not fructose) induced significantly GLUT5 gene expression in comparison to the other dietary treatments in all descendants, being more evident in descendants from control mothers (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eA). Moreover, tagatose (in contrast to fructose) induced the gene expression of the three tagatolytic enzymes in all descendants, although, in this case, the effect was more pronounced and significant in progeny from fructose-fed mothers (Figs.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eB-\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eD).\u003c/p\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003eRegarding the liver, the situation was different. Both sugar transport and its metabolism were preferentially induced by fructose and barely activated by tagatose. In addition, tagatose seemed to be more efficient in affecting its metabolism in the progeny of control mothers than in the progeny of fructose-fed dams (Figs.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eE-\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eH). Thus, fructose activated GLUT5, KHK, ALDOB, and TKFC hepatic gene expression showing a similar profile than the intake of liquid fructose (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eA), that is, in a more evident way in descendants from control dams than in descendants from fructose-fed mothers (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eE-\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eH). Tagatose, however, induced GLUT5 and TFKC, but not KHK and ALDOB, being this effect only found in males from control mothers. Interestingly, tagatolysis was less induced in the intestine of these animals (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eA-\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eD) than in progeny from fructose-fed dams and, therefore, some amount of tagatose is supposed to be able to reach the liver, leading to the effects observed in this tissue (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eE-\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eH).\u003c/p\u003e\u003cp\u003e\u003cb\u003e3.3. Tagatose induced dyslipidemia in the progeny of fructose-fed dams due to an increased intestinal reabsorption and recirculation of bile acids\u003c/b\u003e\u003c/p\u003e\u003cp\u003eAs described in the previous section, tagatose and fructose were absorbed, reaching the systemic circulation and consequently metabolized by the liver and/or intestine. Since tagatose had been ingested in a significant smaller amount than fructose by all descendants, regardless of maternal diet, it was expected that progeny fed tagatose would not show dyslipidaemia. However, surprisingly, this was not the case. Triglyceridemia was significantly modified after the 21 days of nutritional treatment with both fructose and tagatose and, more importantly, this effect was uniquely observed in progeny from fructose-fed dams (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eA). It is widely described that fructose is able to raise plasma triglycerides (\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e), however, tagatose is considered to improve plasma lipid profile (\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e). These results suggest a worsened lipid profile in descendants of fructose-fed mothers consuming liquid fructose or tagatose. To elucidate the underlying mechanisms of this sugar-induced hypertriglyceridemia, and considering the well-established role of fructose in stimulating lipogenesis and inhibiting β-oxidation, we assessed the hepatic expression of key genes involved in fatty acid metabolism, specifically the lipogenic gene stearoyl-CoA desaturase-1 (SCD1) and the catabolic gene carnitine palmitoyl transferase 1 (CPT1) (\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e). Thus, fructose intake produced an overexpression of SCD1 compared to control groups, no matter the mothers' diet, while tagatose consumption did not modify SCD1 gene expression (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eB). This increment in fatty acid synthesis caused by fructose could be providing more substrate for triglycerides production and their exportation to blood, leading to an increase in triglyceridemia. Thus, hepatic gene expression of microsomal triglyceride transfer protein (MTTP) (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eC), a protein involved in very low-density lipoprotein (VLDL) assembly, showed a non-significant trend to increase in males consuming fructose, regardless of the diet of their mothers. Related to fatty acid catabolism, CPT1 gene expression, the enzyme that controls fatty acid entry to mitochondria for its oxidation, tended to be reduced in males from control mothers that were supplemented with fructose, whereas no changes were observed in the corresponding males from fructose-fed mothers (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eD). However, tagatose consumption did not provoke any changes in CPT1 expression in males from control dams, but it produced a non-significant increase in descendants from fructose-fed mothers (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eD). Since tagatose intake did not appear to affect lipogenesis, secretion of triglycerides-rich lipoproteins into the bloodstream or fatty acid oxidation (mechanisms that could account for the tagatose-induced hypertriglyceridemia observed in the offspring of fructose-fed mothers), we evaluated alternative pathways.\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003eOne of these pathways that could affect the lipid profile in blood is the intestinal absorption of lipids. As shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eE, fecal triglycerides remained unchanged among the three groups of descendants from control mothers. In contrast, in the progeny of fructose-fed dams, sugars intake led to a diminution in fecal triglyceride content, which reached statistical significance in the tagatose group when compared to control animals. Consistent with these findings, ileal gene expression of MTTP, the enzyme responsible for assembly in chylomicrons of absorbed lipids, presented no differences in the offspring of control mothers but, importantly, a significant increase was observed in tagatose-supplemented descendants from fructose-fed mothers compared to the control group (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eF).\u003c/p\u003e\u003cp\u003eRelated to that, bile acids are molecules secreted into bile to facilitate intestinal absorption of fats (\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e). In the present study, fecal bile acids did not change in descendants of control mothers after consuming fructose or tagatose compared to the control group (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eA). Interestingly, however, progeny from fructose-fed mothers that received liquid fructose or tagatose did present a non-significant increment in fecal bile acids, being this trend more pronounced in the tagatose group (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eA). Curiously, this same profile was seen in both plasma (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eB) bile acids and hepatic (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eC) bile acids. Thus, in the offspring of fructose-fed mothers that consumed tagatose, plasma bile acids levels were significantly augmented compared to the corresponding control group (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eB) and hepatic bile acids content (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eC) was significantly increased in comparison to both control and fructose groups. These results may indicate an alteration in the enterohepatic bile acid recirculation along the intestine-plasma-liver axis. A higher recirculation of bile acids would lead to an increased intestinal content of bile acids which would facilitate lipids emulsification and absorption. This mechanism could contribute to the lipid alterations observed in the offspring of fructose-fed mothers that consumed tagatose. In accordance with this, all genes involved in bile acid reabsorption in the ileum were overexpressed in the progeny from fructose-fed mothers that consumed tagatose (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eD-G). Thus, whereas in the progeny from control mothers no differences were observed among the three experimental groups, in descendants from fructose-fed mothers, tagatose administration did significantly increase the gene expression of ASBT (apical sodium-bile acid transporter) (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eD) and OST (organic solute transporter), both type alpha (OSTa) (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eF) and type beta (OSTb) (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eG) compared to the control group. In the case of IBABP (ileal bile acid-binding protein) gene expression, the significant increase in the tagatose group was found in comparison to males that consumed fructose (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eE). Moreover, these increases turned out to be also significant for ASBT (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eD) and OSTb (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eG) when comparing males that consumed tagatose from fructose-fed mothers versus descendants of control mothers. Consequently, tagatose intake in descendants from fructose-supplemented mothers would be provoking a higher bile acid entry to the ileum (ASBT), an enhanced bile acid circulation across the enterocyte (IBABP), and a more pronounced bile acid export to blood (OST). These results would highlight how foetal programming induced by maternal fructose intake can modulate the response to a nutrient in progeny.\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003e\u003cb\u003e3.4. Tagatose modified plasma FGF21 but not GLP1 levels in the progeny independently of the mother\u0026acute;s diet.\u003c/b\u003e\u003c/p\u003e\u003cp\u003eIntestinal production of triglyceride-rich lipoproteins has also been shown to be affected by glucagon-like peptide 1 (GLP-1) (\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e), an incretin produced by the gut that has been reported to play a key role in inducing satiety, reducing food intake and controlling obesity. That is why the study of this molecule is attracting increasing interest among the healthcare scientific community. Interestingly, tagatose has been shown to stimulate the release of GLP-1 (\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e), although short-chain fatty acids (SCFA) produced by bacterial fermentation of poorly absorbed sugars into the gut may also stimulate GLP-1 (\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e). In fact, GLP-1 is released in response to the presence of diverse nutrients in the intestine. Expression level of the ileal proglucagon gene (a precursor of GLP-1) has been shown to be increased in the presence of glucose, free fatty acids and SCFA in the distal gut (\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e)\u003c/p\u003e\u003cp\u003eIn the present study, tagatose intake led to an increased ileal gene expression of transporters/receptors of nutrients such as GLUT5 (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eA) for sugars, ABST (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eD) for bile acids, G protein-coupled receptors (GPR41 and GPR43) for SCFAs, and Takeda G protein-coupled receptor (TGR5) for bile acids (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e), being the effect more evident in the progeny of fructose-fed mothers. Interestingly, the upregulation of these genes may help to explain why proglucagon gene expression was induced in descendants of fructose-fed dams consuming tagatose (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003eA), reaching statistical significance in comparison to the control group. However, this increase in proglucagon gene expression in males from fructose-fed mothers consuming tagatose was not accompanied by an augmented expression of PC1/3 (proprotein convertase 1/3) (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003eB), the enzyme responsible for processing proglucagon to the active form of GLP1, nor by a decreased expression of Dpp4 (dipeptidyl peptidase 4), the enzyme that degrades GLP-1 (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003eC). Consequently, plasma GLP1 levels did not differ among all experimental groups (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003eD). It is important to note that we could not measure active GLP-1 levels in serum and the data shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003eD reflect total GLP-1 levels.\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003eIleal and hepatic gene (mRNA) expression of control (C), fructose- (F), and tagatose-supplemented (T) male progeny from control or fructose-fed mothers.\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"9\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c9\" colnum=\"9\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/th\u003e\u003cth align=\"left\" colspan=\"4\" nameend=\"c5\" namest=\"c2\"\u003e\u003cp\u003eCONTROL MOTHERS\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colspan=\"4\" nameend=\"c9\" namest=\"c6\"\u003e\u003cp\u003eFRUCTOSE MOTHERS\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e\u003cb\u003eCONTROL\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e\u003cb\u003eFRUCTOSE\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e\u003cb\u003eTAGATOSE\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003ep\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e\u003cb\u003eCONTROL\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e\u003cb\u003eFRUCTOSE\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e\u003cb\u003eTAGATOSE\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003ep\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colspan=\"9\" nameend=\"c9\" namest=\"c1\"\u003e\u003cp\u003e\u003cb\u003eIleal mRNA Gene Expression (a.u)\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eGPR41\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e1.033\u0026plusmn;0.104\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e1.213\u0026plusmn;0.128\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e1.264\u0026plusmn;0.116\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e0.761\u0026plusmn;0.076\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e0.718\u0026plusmn;0.111#\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e1.255\u0026plusmn;0.192\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e* (FC vs FT)\u003c/p\u003e\u003cp\u003e* (FF vs FT)\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eGPR43\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e1.057\u0026plusmn;0.129\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e1.468\u0026plusmn;0.1778\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e1.637\u0026plusmn;0.205\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e* (CC vs CT)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e1.069\u0026plusmn;0.131\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e1.039\u0026plusmn;0.203\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e1.448\u0026plusmn;0.132\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eTGR5\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e1.132\u0026plusmn;0.087\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e1.78\u0026plusmn;0.178\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e1.687\u0026plusmn;0.263\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e* (CC vs CF)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e1.082\u0026plusmn;0.063\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e1.313\u0026plusmn;0.138\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e1.548\u0026plusmn;0.076\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e** (FC vs FT)\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colspan=\"9\" nameend=\"c9\" namest=\"c1\"\u003e\u003cp\u003e\u003cb\u003eHepatic mRNA Gene Expression (a.u)\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eMCT1\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e1.002\u0026plusmn;0.077\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e1.048\u0026plusmn;0.104\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.977\u0026plusmn;0.043\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e1.158\u0026plusmn;0.112\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e1.044\u0026plusmn;0.056\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e1.335\u0026plusmn;0.127##\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eHDAC1\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e1.024\u0026plusmn;0.08\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e1.003\u0026plusmn;0.136\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e1.290\u0026plusmn;0.105\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e0.975\u0026plusmn;0.047\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e0.868\u0026plusmn;0.121\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e1.062\u0026plusmn;0.095\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eHDAC3\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e0.943\u0026plusmn;0.057\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e0.990\u0026plusmn;0.092\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e1.034\u0026plusmn;0.096\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e1.055\u0026plusmn;0.086\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e1.042\u0026plusmn;0.048\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e0.882\u0026plusmn;0.04\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003ePDK4\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e1.080\u0026plusmn;0.162\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e1.003\u0026plusmn;0.010\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e1.583\u0026plusmn;0.196\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e1.067\u0026plusmn;0.109\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e0.993\u0026plusmn;0.178\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e1.483\u0026plusmn;0.086\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eCIDEC\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e1,008\u0026plusmn;0,048\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e1.105\u0026plusmn;0.108\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e1.360\u0026plusmn;0.089\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e* (CC vs CT)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e1.106\u0026plusmn;0.076\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e1.100\u0026plusmn;0.123\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e1.374\u0026plusmn;0.087\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eVLDLR\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e1.037\u0026plusmn;0.096\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e0.883\u0026plusmn;0.042\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e1.033\u0026plusmn;0.113\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e1.679\u0026plusmn;0.257\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e2.035\u0026plusmn;0.388##\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e1.945\u0026plusmn;0.271##\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eCD36\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e0.0014\u0026plusmn;0.0002\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e0.0016\u0026plusmn;0.0001\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.0024\u0026plusmn;0.0002\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e0.0022\u0026plusmn;0.0006\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e0.0017\u0026plusmn;0.0004\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e0.0011\u0026plusmn;0.0002#\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003ctfoot\u003e\u003ctr\u003e\u003ctd colspan=\"9\"\u003eIleal and hepatic levels of specific mRNA genes are shown. Ileum mRNA expression represents GLP1 signalling pathway genes, and liver mRNA expression represents both SCFA and PPAR alpha signalling pathway genes. Relative target gene mRNA levels were measured by Real Time PCR as explained in Materials and Methods, normalized to Rps29 levels and expressed in arbitrary units (a.u.). Data are means\u0026thinsp;\u0026plusmn;\u0026thinsp;S.E. from 7 to 8 litters. Asterisks denote a significant difference (*, p\u0026thinsp;\u0026lt;\u0026thinsp;0.05; **, p\u0026thinsp;\u0026lt;\u0026thinsp;0.01; ***, p\u0026thinsp;\u0026lt;\u0026thinsp;0.001) between the groups with a different diet but the same mothers ́ diet. Hash symbols denote a significant difference (#, p\u0026thinsp;\u0026lt;\u0026thinsp;0.05; ##, p\u0026thinsp;\u0026lt;\u0026thinsp;0.01; ###, p\u0026thinsp;\u0026lt;\u0026thinsp;0.001) as compared to the control mothers (groups with the same diet but different mothers ́ diet). fructose. The first letter indicates whether the mothers had been supplied with tap water during pregnancy (C: control) or liquid fructose (F); and the second letter indicates the nutritional treatment without (C: control) or with additives, fructose (F) or tagatose (T), when they were adults. GLP1: glucagon-like protein 1; and GLP1 signalling: GPR: G protein-coupled receptors; TGR: Takeda G protein-coupled receptor. SCFA: short-chain fatty acids; and SCFA signalling: MCT1: monocarboxylate transporter 1 protein; HDAC: Histone deacetylases; PPAR: peroxisome proliferator-activated receptor; and PPAR signalling: PDK: pyruvate dehydrogenase kinase; CIDEC: cell death-inducing DNA fragmentation factor-like effector C; VLDLR: very low-density lipoprotein receptor; CD36: cluster of differentiation 36.\u003c/td\u003e\u003c/tr\u003e\u003c/tfoot\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003eOn the other hand, it has been reported that FGF21, a hormone that also responds to nutrients, can regulate sugar preference and, in addition, its production is induced by GLP-1 analogues (\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e). In the present work we found that FGF21 plasma levels were augmented in sugar-supplemented descendants, regardless of the diet consumed by their mothers (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003eE). Curiously, plasma FGF21 levels paralleled the different amount of liquid fructose ingested by descendants, which was influenced by their mothers\u0026acute;diet (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eA). Importantly, tagatose-fed descendants, who had consumed approximately 10 times less amount of sugar than the fructose-fed groups, showed elevated plasma FGF21 levels. This increase in the tagatose-supplemented animals was significant compared to the control group in the progeny of control mothers and compared to the control and fructose-fed groups in descendants from fructose-fed dams (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003eE). Considering that FGF21 is mainly synthesized in the liver and that our results indicate that tagatose, unlike fructose, barely reaches this organ, it was very striking to find that tagatose intake induced hepatic FGF21 gene expression whereas fructose did not. Moreover, this tagatose-induced activation was more pronounced in the offspring of fructose-fed dams than in those of control mothers (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003eF).\u003c/p\u003e\u003cp\u003e\u003cb\u003e3.5. Tagatose intake increased plasma levels of Ang II which produced worse effects in the offspring of fructose-fed mothers.\u003c/b\u003e\u003c/p\u003e\u003cp\u003eIt has been shown that, in response to carbohydrate intake, the liver produces FGF21 which acts on the hypothalamus to selectively suppress sugar intake. Interestingly, in the present study, after tagatose consumption, plasma FGF21 levels and sugar intake were inversely related, that is, tagatose increased plasma FGF21, and this reduced sugar appetite (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003eE versus Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eA). In contrast, in fructose-fed animals we found a direct relationship between sugar intake and FGF21 levels. Carbohydrates are known to activate hepatic ChREBP, a transcription factor that promotes FGF21 production in the liver (\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e). However, as previously mentioned, tagatose consumption hardly affected the hepatic expression of some ChREBP target genes such as KHK and ALDOB (Figs.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eF and \u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eG). These data suggest that ChREBP is likely not involved in the tagatose-induced activation of hepatic FGF21 production.\u003c/p\u003e\u003cp\u003eTagatose is minimally absorbed in the intestine and, thus, it has been proposed as a prebiotic agent capable of selectively stimulating the growth of specific gut microbiota and affecting host health (\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e). By doing this, tagatose induces microbiota that produce beneficial compounds such as butyrate. This SCFA has been demonstrated to stimulate hepatic FGF21 gene expression by inhibiting histone deacetylase 3 (HDAC3) which suppresses the activity of peroxisome proliferator-activated receptor type alpha (PPARα) (\u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e). And, to note, FGF21 is a well-known PPAR alpha target gene in liver (\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e). However, none of the nutritional interventions here used altered the gene expression of monocarboxylate transporter 1 (MCT1), which permits SCFA entry to the cell, or that of HDAC1 or HDAC3 (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). Thus, exploring the role of PPARα in the tagatose-induced activation of FGF21, we found that whereas CPT1 (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eD), PDK4 and CIDEC gene expression (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e) observed in the tagatose groups could reflect an activation of PPARα, the findings observed for SCD1 (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eB), VLDLR and CD36 gene expression did not show changes in that sense (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). Moreover, it has been suggested that part of the effect of PPARα on hepatic ketogenesis may be mediated by induction of the PPARα target FGF21; however, plasma ketone bodies trended to be diminished in all descendants consuming carbohydrates regardless of their mothers\u0026acute;diet [166.1\u0026thinsp;\u0026plusmn;\u0026thinsp;28.4; 92.9\u0026thinsp;\u0026plusmn;\u0026thinsp;19.3; and 95.6\u0026thinsp;\u0026plusmn;\u0026thinsp;5.1 for control, fructose- and tagatose-fed descendants from control mothers; 206.3\u0026thinsp;\u0026plusmn;\u0026thinsp;41.2; 143.1\u0026thinsp;\u0026plusmn;\u0026thinsp;41.1; and 165.7\u0026thinsp;\u0026plusmn;\u0026thinsp;52.3 \u0026micro;M for control, fructose- and tagatose-fed males from fructose-fed dams].\u003c/p\u003e\u003cp\u003eImportantly, angiotensin II (Ang II), a molecule that promotes inflammation, oxidative stress, vascular injury, fatty liver and insulin resistance (\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e), has been shown to be able to increase serum FGF21 levels and hepatic FGF21 gene expression, possibly as a compensatory and protective response against these harmful effects induced by Ang II (\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e, \u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e38\u003c/span\u003e). As shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003eA, and in accordance with the results found in hepatic FGF21 gene expression (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003eG), tagatose intake produced an increase in plasma Ang II in both descendants from control mothers and males from fructose-fed dams. This augmentation observed in tagatose-fed progeny turned out to be significant versus fructose-fed animals in descendants of control mothers and versus the control group in males from fructose-fed mothers. In accordance, the angiotensin-converting enzyme (ACE), that converts AngI to AngII, although its hepatic gene expression was not affected by any dietary treatment (Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003eB), ileal ACE gene expression was significantly augmented after carbohydrate ingestion in males from control mothers and sharply increased after tagatose intake in progeny from fructose-fed dams. In fact, these tagatose-mediated effects were significant when compared to the other two groups of descendants from fructose-fed dams. Moreover, tagatose-supplemented males from fructose-fed mothers showed a significant higher expression of ileal ACE than the descendants of control mothers fed with the same sugar (denoted by a hash symbol) suggesting a fetal programming mediated effect (Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003eC).\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003eAdditionally, Ang II is well-known to stimulate vasopressin secretion (\u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e39\u003c/span\u003e), a molecule that is related to liquid ingestion, carbohydrate (fructose, glucose or HFCS) intake and it has been proposed as a key mediator in the fructose-induced metabolic syndrome (\u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e40\u003c/span\u003e). However, we measured plasma copeptin, an analogue and stable peptide derived from the precursor of vasopressin, and no differences were found among all the experimental groups [46.1\u0026thinsp;\u0026plusmn;\u0026thinsp;6.8; 36.8\u0026thinsp;\u0026plusmn;\u0026thinsp;5.4; and 49.9\u0026thinsp;\u0026plusmn;\u0026thinsp;4.9 for control, fructose- and tagatose-fed descendants from control mothers; 49.4\u0026thinsp;\u0026plusmn;\u0026thinsp;6.9; 46.0\u0026thinsp;\u0026plusmn;\u0026thinsp;0.8; and 39.9\u0026thinsp;\u0026plusmn;\u0026thinsp;3.0 pg/mL for control, fructose- and tagatose-fed males from fructose-fed dams]. Nevertheless, despite this lack of effect mediated by carbohydrate intake in copeptin levels, the mRNA gene expression of vasopressin 1a receptor (AVP1AR) showed a reduction in the liver of fructose-fed animals from control mothers in consonance to previous studies (\u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e40\u003c/span\u003e) and a significant reduction in tagatose-supplemented males from these same mothers (Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003eD). Curiously, in the progeny from fructose-fed mothers, the effect produced by fructose ingestion was not observed whereas tagatose intake significantly diminished AVP1AR expression versus both control and fructose groups. Furthermore, this fetal programming effect was more evident in heart AVP1AR expression (Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003eE), since no changes were observed among the three experimental groups of descendants from control mothers, but a significant reduction was found in tagatose-fed males from fructose-fed mothers in comparison to the other two groups and also when compared to the corresponding tagatose-fed group from control mothers (hash symbol). Interestingly, Andres-Hernando et al observed in AVP1AR-KO mice worse metabolic features of metabolic syndrome than in the wild-type group (\u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e40\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eIn view of these interesting results observed in tagatose-fed males, that is, elevated levels of plasma FGF21 trying to compensate the high levels of plasma Ang II and a lower presence of the vasopressin 1a receptor, we decided to evaluate oxidative stress and lipid and glucose dysfunction parameters in both liver and heart. Curiously, FGF21 appeared to counteract the effects of Ang II in the liver but not in the heart. This protective effect was more evident in the progeny from control mothers than in those from fructose-fed dams. Thus, in the liver, none of the nutritional interventions altered MDA levels, catalase activity or superoxide dismutase activity, regardless of the maternal diet (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). In contrast, in the heart, there was a tendency for these oxidative markers to be modified by carbohydrate intake in progeny from control mothers, although statistically significant differences were only found in tagatose-fed descendants from fructose-fed mothers. Specifically, catalase activity was significantly decreased compared to the fructose group, while SOD activity was elevated versus both fructose and control groups (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). As shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig7\" class=\"InternalRef\"\u003e7\u003c/span\u003eA, whereas liver glycogen content remained unchanged in all experimental groups, cardiac glycogen content (Fig.\u0026nbsp;\u003cspan refid=\"Fig7\" class=\"InternalRef\"\u003e7\u003c/span\u003eB) was augmented by carbohydrate intake in the progeny from control mothers, becoming significantly different in tagatose-fed males versus the control group. This tagatose-mediated increase in cardiac glycogen was even more pronounced and significant in descendants from fructose-fed dams versus the other two groups. Additionally, a similar profile to the one observed in oxidative parameters and glycogen was also found in the triglycerides content. Thus, hepatic steatosis was not observed in any case, although a slight non-significant lipid accretion was found in tagatose-fed animals from fructose-fed dams (Fig.\u0026nbsp;\u003cspan refid=\"Fig7\" class=\"InternalRef\"\u003e7\u003c/span\u003eC). Interestingly, whereas cardiac triglycerides content did not change among the three experimental groups in males from control dams, a significant triglycerides deposit was found in males from fructose-fed dams after consuming tagatose (Fig.\u0026nbsp;\u003cspan refid=\"Fig7\" class=\"InternalRef\"\u003e7\u003c/span\u003eD) compared to the control group.\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003eHepatic and cardiac oxidative stress parameters of control (C), fructose- (F), and tagatose-supplemented (T) male progeny from control or fructose-fed mothers. Data are means\u0026thinsp;\u0026plusmn;\u0026thinsp;S.E. from 7 to 8 litters. Asterisks denote a significant difference (*, p\u0026thinsp;\u0026lt;\u0026thinsp;0.05; **, p\u0026thinsp;\u0026lt;\u0026thinsp;0.01; ***, p\u0026thinsp;\u0026lt;\u0026thinsp;0.001) between the groups with a different diet but the same mothers ́ diet. Hash symbols denote a significant difference (#, p\u0026thinsp;\u0026lt;\u0026thinsp;0.05; ##, p\u0026thinsp;\u0026lt;\u0026thinsp;0.01; ###, p\u0026thinsp;\u0026lt;\u0026thinsp;0.001) as compared to the control mothers (groups with the same diet but different mothers ́ diet). fructose. The first letter indicates whether the mothers had been supplied with tap water during pregnancy (C: control) or liquid fructose (F); and the second letter indicates the nutritional treatment without (C: control) or with additives, fructose (F) or tagatose (T), when they were adults. MDA: malondialdehyde; Cat: catalase activity; SOD: superoxide dismutase activity.\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"11\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c9\" colnum=\"9\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c10\" colnum=\"10\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c11\" colnum=\"11\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/th\u003e\u003cth align=\"left\" colspan=\"4\" nameend=\"c5\" namest=\"c2\"\u003e\u003cp\u003eCONTROL MOTHERS\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colspan=\"5\" nameend=\"c10\" namest=\"c6\"\u003e\u003cp\u003eFRUCTOSE MOTHERS\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colspan=\"1\" nameend=\"c11\" namest=\"c11\"\u003e\u0026nbsp;\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e\u003cb\u003eCONTROL\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e\u003cb\u003eFRUCTOSE\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e\u003cb\u003eTAGATOSE\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003ep\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e\u003cb\u003eCONTROL\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e\u003cb\u003eFRUCTOSE\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e\u003cb\u003eTAGATOSE\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003ep\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colspan=\"2\" nameend=\"c11\" namest=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colspan=\"11\" nameend=\"c11\" namest=\"c1\"\u003e\u003cp\u003e\u003cb\u003eLiver oxidative-stress parameters\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eMDA (mmol/g tissue)\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e15.552\u0026plusmn;1.357\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e15.398\u0026plusmn;1.737\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e17.383\u0026plusmn;1.616\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e14.553\u0026plusmn;0.463\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e14.447\u0026plusmn;0.664\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e18.227\u0026plusmn;1.924\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colspan=\"2\" nameend=\"c11\" namest=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eCat (mU/mg prot)\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e1391.6\u0026plusmn;120.8\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e1036.4\u0026plusmn;103.7\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e1373.4\u0026plusmn;117.8\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e1578.1\u0026plusmn;121.5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e1682.3\u0026plusmn;190.1##\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e1663.9\u0026plusmn;232.6\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colspan=\"2\" nameend=\"c11\" namest=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eSOD (U/mg prot)\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e43.33\u0026plusmn;2.67\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e42.37\u0026plusmn;1.43\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e44.35\u0026plusmn;1.67\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e40.79\u0026plusmn;0.47\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e43.11\u0026plusmn;0.33\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e44.65\u0026plusmn;2.32\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colspan=\"2\" nameend=\"c11\" namest=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colspan=\"11\" nameend=\"c11\" namest=\"c1\"\u003e\u003cp\u003e\u003cb\u003eHeart oxidative-stress parameters\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eMDA (nmol/g tissue)\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e36.17\u0026plusmn;4.21\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e52.33\u0026plusmn;7.34\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e57.87\u0026plusmn;12.77\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e34.38\u0026plusmn;4.53\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e48.70\u0026plusmn;12.83\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e38.55\u0026plusmn;5.64\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colspan=\"2\" nameend=\"c11\" namest=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eCat (mU/mg prot)\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e42.91\u0026plusmn;2.65\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e39.52\u0026plusmn;2.42\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e36.07\u0026plusmn;2.20\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e40.67\u0026plusmn;1.08\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e43.48\u0026plusmn;1.90\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e33.82\u0026plusmn;1.54\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e** (FF vs FT)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colspan=\"2\" nameend=\"c11\" namest=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eSOD (U/mg prot)\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e23.54\u0026plusmn;0.52\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e24.57\u0026plusmn;0.49\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e27.04\u0026plusmn;1.38\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e23.78\u0026plusmn;0.92\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e23.42\u0026plusmn;1.51\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e28.83\u0026plusmn;1.21\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e* (FC vs FT)\u003c/p\u003e\u003cp\u003e** (FF vs FT)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colspan=\"2\" nameend=\"c11\" namest=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003eTherefore, this oxidative stress and accumulation of fuel stores suggest that tagatose consumption induces cardiac metabolic dysregulation in the offspring of fructose-fed mothers (\u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e41\u003c/span\u003e, \u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e42\u003c/span\u003e) that ultimately would lead to cardiac dysfunction. Moreover, we have previously described a similar scenario in which FGF21 was able to protect against lipid accretion and oxidative stress influenced by maternal nutrition and in a tissue-dependent manner (\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e).\u003c/p\u003e\u003c/div\u003e"},{"header":"4. DISCUSSION","content":"\u003cp\u003eThe urgent need to reduce the high prevalence of metabolic diseases, especially those driven by modifiable lifestyle factors such as \"diseases related to processed food\", makes dietary changes essential. One key aspect of these changes involves reducing added sugars intake, particularly fructose or its derivatives, and replacing them with alternative sugars or sweeteners, such as rare sugars. This shift has already been taking place since the first decades of this century and, fortunately, it has contributed to a gradual slowdown in the development of metabolic diseases. However, the decrease in added sugar consumption has not been paralleled by a proportional reduction in the incidence of metabolic diseases such as obesity (\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e). We believe this discrepancy is because high fructose consumption was common among pregnant women prior to these dietary changes. Consequently, it is logical that the offspring of these mothers continue to exhibit a high incidence of metabolic diseases and that their response to diet is affected by a fetal programming mechanism due to the maternal intake of added sugar (\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e). Given the increasing incorporation of rare sugars to replace fructose, sucrose or HFCS in our diet, we decided to find out the impact of maternal fructose intake on offspring\u0026acute;s metabolic response to consumption of the rare sugar tagatose.\u003c/p\u003e\u003cp\u003eOne of the most striking results found is that, despite the low intake of tagatose (about 5% of total caloric intake), it was able to stimulate the gene expression of enzymes and transporters of the tagatolysis pathway in the intestine, in a more pronounced way in descendants of fructose-fed mothers, and, interestingly, with almost no influence on hepatic tagatolysis. In contrast, fructose, which accounted for about 45% of total caloric intake, had the opposite effect, that is, it did not modify intestinal fructolysis and clearly affected the gene expression of liver fructolysis enzymes and transporters (\u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e43\u003c/span\u003e). These data suggest that the effects observed mainly in the offspring of fructose-fed mothers are more related to a metabolite produced by intestinal tagatolysis than to tagatose itself.\u003c/p\u003e\u003cp\u003eAnother relevant finding was that tagatose intake affected triglyceridemia in a way that was clearly influenced by fetal programming, since it induced hypertriglyceridemia in the offspring of fructose-fed mothers, but not in those of control mothers. Fructose intake also elevated triglyceridemia in that same group of descendants. Interestingly, while fructose-mediated effect was mainly caused by a higher expression of lipogenic genes in the liver, the effect of tagatose appeared to be associated with a higher gene expression of bile acid transporters in the intestine. This result suggests a greater enterohepatic recirculation of bile acids, which led to a greater lipid absorption and packaging of in the intestine. Such a mechanism would explain the hyperlipidemia observed in these descendants and aligns with findings previously described by us and others although using another type of diet (\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e, \u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e44\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eInterestingly, gene expression of most transporters and receptors of nutrients, such as sugars, short-chain fatty acids, and bile acids, was increased in the ileum after tagatose intake, particularly in the offspring of fructose-fed dams. These transporters and receptors have been directly related to intestinal production of proglucagon in response to these nutrients (\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e). In fact, consistent with this, we observed that the tagatose-induced upregulation of the expression of nutrient transporters and receptors was accompanied by an increase in the expression of proglucagon. However, plasma GLP1 levels did not reflect this observed rise in its precursor molecule. In addition, GLP2, which is also produced from proglucagon and whose high levels would be more in line with the greater lipid absorption and packaging found in the descendants of fructose-fed mothers consuming tagatose, was not affected by the different treatments (\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e). Unfortunately, total GLP was measured and not the active form that would be more informative.\u003c/p\u003e\u003cp\u003eOne of the most striking results was to find that both tagatose intake and, as expected, fructose consumption increased plasma levels of FGF21 (\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e). Interestingly, although tagatose (as mentioned above) seemed to barely reach the liver as it failed to stimulate tagatolysis in this organ, it was able to clearly increase the hepatic expression of FGF21. FGF21 has been described as a molecule capable of decreasing the sweet taste preference (\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e). Therefore, it was paradoxical to find that, whereas fructose intake was directly related to plasma FGF21 levels, the relationship with tagatose consumption was clearly opposite and more in line with what has been previously described, that is, the tagatose-induced increase in FGF21 could be the cause of the low intake of this sugar observed in the offspring.\u003c/p\u003e\u003cp\u003eWe were therefore interested in discovering the mechanism by which tagatose intake increases the hepatic expression of FGF21. We found that classic effectors such as ChREBP, PPAR alpha and even the mediation of SCFA that could be produced in the intestine by unabsorbed tagatose, failed to explain it adequately (\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e). However, a less studied regulatory molecule in the metabolism of FGF21, such as angiotensin II, was found to be increased in plasma after tagatose intake. This increase was consistent with the findings observed in hepatic expression of FGF21, so we could affirm that tagatose consumption increased Ang II levels and, with it, hepatic production of FGF21.\u003c/p\u003e\u003cp\u003eInterestingly, Ang II has also been shown to be involved in the production of vasopressin (\u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e39\u003c/span\u003e), a molecule that could influence the lower fluid intake seen in animals that consumed tagatose, as has been described in the case of fructose (\u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e40\u003c/span\u003e). However, levels of copeptin (a stable form of vasopressin) were not changed by any of the treatments. On the other hand, the hepatic expression of its receptor 1a (AVP1AR) was modified by fructose intake, which decreased it, confirming previous findings by other authors (\u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e40\u003c/span\u003e). Notably, tagatose reduced the hepatic gene expression of this receptor more drastically than fructose in all descendants and, furthermore, it also diminished AVP1AR gene expression in the heart, although in this case this effect was only observed in descendants of fructose-mothers. Considering that the lack of this receptor has been related to the appearance of more severe symptoms of metabolic syndrome, this interesting result warrants further investigation in future studies (\u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e40\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eIt has been described that Ang II promotes the production of FGF21 to counteract its adverse effects (\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e, \u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e38\u003c/span\u003e). Thus, in the present study, FGF21 manages to counteract the negative effects of Ang II in the liver of all descendants fed tagatose. However, this was not the case in the heart of males from fructose-fed mothers since oxidative stress and a clear accumulation of glycogen and triglycerides were observed after the intake of tagatose. These dysfunctions could be an initial biomarker of cardiac metabolic dysregulation, a very common situation observed in diabetes or metabolic syndrome (\u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e41\u003c/span\u003e).\u003c/p\u003e"},{"header":"5. CONCLUSIONS","content":"\u003cp\u003eTherefore, the consumption of alternative sugars to fructose and/or HFCS, such as tagatose, by descendants of mothers who consumed fructose during their pregnancy could lead to adverse effects more likely than in descendants of control mothers.\u003c/p\u003e\u003cp\u003eThe current study highlights a novel interaction between tagatose consumption and angiotensin II signaling, leading to increased hepatic FGF21 expression even in the absence of direct hepatic sugar metabolism. This unique metabolic response raises concerns regarding its systemic effects, particularly in individuals with prenatal exposure to excess fructose. The inability of FGF21 to mitigate Ang II-associated dysfunction in cardiac tissue suggests early biomarkers of metabolic disruption. These data call for a re-evaluation of the metabolic safety of rare sugars like tagatose.\u003c/p\u003e\u003cp\u003eOverall, these findings emphasize the key role of maternal nutrition during gestation, particularly the quality and quantity of sugar intake, as it can seriously affect the long-term metabolic health of the progeny through fetal programming mechanisms. Consequently, these observations along with previous reports strongly support the adherence to the WHO guidelines which recommend limiting the intake of simple sugars in processed foods and sugary drinks to less than 10% of total daily caloric intake. Importantly, as evidenced by the present study, such limitations should also include rare sugars such as tagatose.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAcknowledgements\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors thank Jose M. Garrido and his team for their help in handling the rats.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthor contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eC.B. and E.F. conceived and designed the study. E.F., M.P-A., C.D., P.O. and M.I.P. contributed reagents/materials/analysis tools for gene expression studies and parameter analysis. E.F., P.O. and M.I.P. handled the animals. M.I.P. analysed the data. C.B. and E.F. wrote the paper. All authors had access to the study data and had reviewed and approved the final manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis work was supported by funds from the Ministerio de Ciencia e Innovación (MCIN): PID2020-118054RB-I00/AEI/10.13039/501100011033 and PID2023-152756OB-I00/AEI/10.13039/501100011033. Elena Fauste was previously supported, and Madelín Pérez-Armas is supported with FPU fellowships from MCIN.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAvailability of data and materials\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll data generated or analysed during this study are included in this published article [and its supplementary information files].\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthics approval and consent to participate\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe experimental protocol was approved by the Ethical Committee for Animal Experimentation of the University San Pablo-CEU and by Autonomous Government of Madrid (ref. numbers 10/206458.9/13 and 10/042445.9/19).\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\u003eCompeting interests\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThere are no conflicts of interest to declare.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eCarrera-Bastos P, Fontes-Villalba M, O\u0026rsquo;Keefe JH, Lindeberg S, Cordain L. The western diet and lifestyle and diseases of civilization. Res Rep Clin Cardiol. 2011;2(1):15\u0026ndash;35.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eKopp W. How Western Diet And Lifestyle Drive The Pandemic Of Obesity And Civilization Diseases. Diabetes Metab Syndr Obes. 2019;12:2221\u0026ndash;36.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eHowie GJ, Sloboda DM, Kamal T, Vickers MH. Maternal nutritional history predicts obesity in adult offspring independent of postnatal diet. J Physiol. 2009;587(Pt 4):905\u0026ndash;15.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eWhite JS, Nicklas TA. High-fructose corn syrup use in beverages: composition, manufacturing, properties, consumption, and health effects. Beverage impacts on health and nutrition: Springer; 2016. pp. 285\u0026ndash;301.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eAlwahsh SM, Gebhardt R. 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Clin Chem. 1987;33(2):214\u0026ndash;20.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eAebi H. [13] Catalase in vitro. Methods in enzymology. Volume 105. Elsevier; 1984. pp. 121\u0026ndash;6.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eYe J, Coulouris G, Zaretskaya I, Cutcutache I, Rozen S, Madden TL. Primer-BLAST: a tool to design target-specific primers for polymerase chain reaction. BMC Bioinformatics. 2012;13:134.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003ePfaffl MW. A new mathematical model for relative quantification in real-time RT\u0026ndash;PCR. Nucleic Acids Res. 2001;29(9):e45\u0026ndash;e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eAidoo RP, Depypere F, Afoakwa EO, Dewettinck K. Industrial manufacture of sugar-free chocolates\u0026ndash;Applicability of alternative sweeteners and carbohydrate polymers as raw materials in product development. 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Curr Opin Lipidol. 2018;29(2):95\u0026ndash;103.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eDantas Machado AC, Brown SD, Lingaraju A, Sivaganesh V, Martino C, Chaix A, et al. Diet and feeding pattern modulate diurnal dynamics of the ileal microbiome and transcriptome. Cell Rep. 2022;40(1):111008.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eKim KH, Lee MS. FGF21 as a mediator of adaptive responses to stress and metabolic benefits of anti-diabetic drugs. J Endocrinol. 2015;226(1):R1\u0026ndash;16.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003evon Holstein-Rathlou S, BonDurant LD, Peltekian L, Naber MC, Yin TC, Claflin KE, et al. FGF21 Mediates Endocrine Control of Simple Sugar Intake and Sweet Taste Preference by the Liver. Cell Metab. 2016;23(2):335\u0026ndash;43.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eSon S, Koh J, Park M, Ryu S, Lee W, Yun B, et al. Effect of the Lactobacillus rhamnosus strain GG and tagatose as a synbiotic combination in a dextran sulfate sodium-induced colitis murine model. J Dairy Sci. 2019;102(4):2844\u0026ndash;53.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eLi H, Gao Z, Zhang J, Ye X, Xu A, Ye J, et al. Sodium butyrate stimulates expression of fibroblast growth factor 21 in liver by inhibition of histone deacetylase 3. Diabetes. 2012;61(4):797\u0026ndash;806.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eRakhshandehroo M, Knoch B, M\u0026uuml;ller M, Kersten S. Peroxisome proliferator-activated receptor alpha target genes. PPAR Res. 2010;2010.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eMastoor Z, Diz-Chaves Y, Gonz\u0026aacute;lez-Mat\u0026iacute;as LC, Mallo F. Renin-Angiotensin System in Liver Metabolism: Gender Differences and Role of Incretins. 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Vasopressin mediates fructose-induced metabolic syndrome by activating the V1b receptor. JCI Insight. 2021;6(1).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eVarma U, Koutsifeli P, Benson V, Mellor K, Delbridge L. Molecular mechanisms of cardiac pathology in diabetes\u0026ndash;Experimental insights. Biochimica et Biophysica Acta (BBA)-Molecular Basis of Disease. 2018;1864(5):1949\u0026ndash;59.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eYe G, Metreveli NS, Donthi RV, Xia S, Xu M, Carlson EC, et al. Catalase protects cardiomyocyte function in models of type 1 and type 2 diabetes. Diabetes. 2004;53(5):1336\u0026ndash;43.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eAndres-Hernando A, Orlicky DJ, Kuwabara M, Ishimoto T, Nakagawa T, Johnson RJ, et al. Deletion of Fructokinase in the Liver or in the Intestine Reveals Differential Effects on Sugar-Induced Metabolic Dysfunction. Cell Metab. 2020;32(1):117\u0026ndash;e273.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eDowning LE, Edgar D, Ellison PA, Ricketts ML. Mechanistic insight into nuclear receptor-mediated regulation of bile acid metabolism and lipid homeostasis by grape seed procyanidin extract (GSPE). Cell Biochem Funct. 2017;35(1):12\u0026ndash;32.\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"molecular-medicine","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"mome","sideBox":"Learn more about [Molecular Medicine](https://molmed.biomedcentral.com)","snPcode":"10020","submissionUrl":"https://submission.springernature.com/new-submission/10020/3","title":"Molecular Medicine","twitterHandle":"@MolecularMedic1","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"BMC/SO AJ","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"Fructose, pregnancy, foetal programming, bile acids, FGF21, Angiotensin II, tagatose","lastPublishedDoi":"10.21203/rs.3.rs-7489857/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7489857/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cem\u003e\u003cstrong\u003eBackground:\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e \u003c/strong\u003eMaternal fructose intake induces harmful effects in progeny. However, this sugar is not contraindicated during pregnancy. On the other hand, the use of low-calorie sweeteners, such as tagatose, is increasing. Thus, we have studied whether the consumption of tagatose compared to fructose affects lipid metabolism in the offspring of mothers which were supplemented with fructose during their pregnancy.\u003c/p\u003e\n\u003cp\u003e\u003cem\u003e\u003cstrong\u003eMethods:\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e \u003c/strong\u003eThree-month-old male offspring from control or fructose mothers received liquid 10% fructose or tagatose for 21 days. A control group (without any additive) was also included. Biochemical and molecular parameters were determined in plasma, tissues and feces.\u003c/p\u003e\n\u003cp\u003e\u003cem\u003e\u003cstrong\u003eResults:\u003c/strong\u003e\u003c/em\u003e\u003cem\u003e \u003c/em\u003eBoth tagatose and fructose consumption caused hypertriglyceridemia in descendants of fructose-fed mothers. Whereas fructose consumption led to a greater hepatic lipogenesis, tagatose supplementation provoked a higher enterohepatic bile acids recirculation, and therefore a higher intestinal lipid absorption and assembly. However, plasma GLP1, a molecule that affects lipid intestinal absorption, was unchanged. Curiously, FGF21, a molecule which regulates lipid and carbohydrate metabolism and is sensitive to GLP1, was augmented in plasma and liver of tagatose-supplemented descendants regardless of their maternal diet. Interestingly, Angiotensin II (Ang II), which can induce FGF21 production, was increased in plasma of all animals supplemented with tagatose. However, the deleterious effects of Ang II were effectively reversed by FGF21 in males from control mothers, but not in descendants of fructose-fed dams.\u003c/p\u003e\n\u003cp\u003e\u003cem\u003e\u003cstrong\u003eConclusions:\u003c/strong\u003e\u003c/em\u003e Maternal fructose consumption determines the response of the offspring to tagatose intake, causing an increased intestinal lipid absorption, and metabolic changes that are characteristic of metabolic syndrome such as dyslipidaemia, steatosis and oxidative stress.\u003c/p\u003e","manuscriptTitle":"Tagatose Consumption Provokes Metabolic Syndrome Features in Rat Males from Mothers That Consumed Fructose During Their Pregnancy","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-09-29 16:37:28","doi":"10.21203/rs.3.rs-7489857/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2025-10-07T19:27:41+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-10-07T19:06:29+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-10-04T11:21:40+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-09-30T02:04:11+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-09-23T15:43:03+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"332768662636058635351900852395806522592","date":"2025-09-23T13:46:33+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"103225659285846451555637010256176967929","date":"2025-09-22T03:56:11+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"48921920769443914246797887503739092653","date":"2025-09-20T13:25:56+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"78425371400523510030854341412565113002","date":"2025-09-18T23:59:20+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2025-09-18T13:41:48+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2025-09-02T01:29:27+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2025-09-02T01:29:18+00:00","index":"","fulltext":""},{"type":"submitted","content":"Molecular Medicine","date":"2025-08-29T15:13:48+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"molecular-medicine","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"mome","sideBox":"Learn more about [Molecular Medicine](https://molmed.biomedcentral.com)","snPcode":"10020","submissionUrl":"https://submission.springernature.com/new-submission/10020/3","title":"Molecular Medicine","twitterHandle":"@MolecularMedic1","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"BMC/SO AJ","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"68f107e1-d4c3-4a1a-92d5-ec34aa4ab0d4","owner":[],"postedDate":"September 29th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[],"tags":[],"updatedAt":"2026-01-05T16:02:51+00:00","versionOfRecord":{"articleIdentity":"rs-7489857","link":"https://doi.org/10.1186/s10020-025-01402-3","journal":{"identity":"molecular-medicine","isVorOnly":false,"title":"Molecular Medicine"},"publishedOn":"2025-12-29 15:57:42","publishedOnDateReadable":"December 29th, 2025"},"versionCreatedAt":"2025-09-29 16:37:28","video":"","vorDoi":"10.1186/s10020-025-01402-3","vorDoiUrl":"https://doi.org/10.1186/s10020-025-01402-3","workflowStages":[]},"version":"v1","identity":"rs-7489857","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7489857","identity":"rs-7489857","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}