Effect of fecal microbiota transplantation on “intestinal flora-SCFAs-GPR43 - gastrointestinal peptide” pathway in rats with high-fat diet

Effect of fecal microbiota transplantation on “intestinal flora-SCFAs-GPR43 - gastrointestinal peptide” pathway in rats with high-fat diet | 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 Article Effect of fecal microbiota transplantation on “intestinal flora-SCFAs-GPR43 - gastrointestinal peptide” pathway in rats with high-fat diet He Yu, Lijun Cui, Xiaomei Wang, Xu Wang, Shaoyang Liu, Chen Bai, and 3 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6307329/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Objective To explore the effect of high-fat diet and fecal microbiota transplantation (FMT) on “intestinal flora-SCFAs-GPR43-gastrointestinal peptide” pathway, and provide evidence and clues for the prevention and treatment of obesity caused by eutrophic diet. Methods 160 male SD rats were used in this study, 50 of them were randomly selected to be fed a standard rat diet, while the remaining 110 rats were fed a high-fat diet (D12492). After excluding the rats that did not meet the obesity criteria, the remaining rats were subjected to treatment with normal microbiota enema and obesity-associated microbiota enema. The rats were divided into normal control group 1 (NC1), normal control group 2 (NC2), obesity model group (M), obesity fecal microbiota transplantation group (FMT1), and normal fecal microbiota transplantation group (FMT2). The study observed the general situation, the index of liver, spleen and thymus in rats. Morphological changes of colon and liver tissues were examined under an optical microscope, and the alterations in gut microbiota were detected by 16s rDNA. Gas chromatography-mass spectrometry (GC-MS) was ued to measure the levels of short-chain fatty acids (SCFAs), including acetic acid, propionic acid, and butyric acid. Immunohistochemistry (IHC) was used to evaluate the expression of GPR43 in liver tissue. Additionally, gastrointestinal peptides in rat serum were quantified using the ELISA method, while cholesterol and triglyceride levels in serum were measured using an automatic biochemical analyzer. Results The high-fat diet successfully induced obesity rat models. This led to significant changes in gut environment and the survival environment of microbiota, such as Lactobacillus, reflecting the intestinal microecological disorders in rats with high-fat diet induced obesity, Different dietary interventions can lead to varing developments in gut microbiota. After antibiotic intervention, gut microbiota in rats were significantly suppressed, with reduced species diveristy and abundance, establishing an antibiotic-induced rat model. High-fat diet interventions resulted in significant changes in the relative abundances of specific gut bacterial species. Further analysis of microbial metabolites displayed that a high-calorie diet reduced the content of short-chain fatty acids (SCFAs) in feces, and subsequently reduced the expression of GPR43, resulting in improved abnormal expression of downstream gastrointestinal peptide. Conclusions High-fat diet affects the intestinal flora-SCFAs-GPR43-gastrointestinal peptide pathway, leading to related pathological reactions, such as intestinal flora imbalance and short-chain fatty acid metabolism disorders, which in return activates GPR43, and releases PYY, GLP-1, GAS, MTL, causing lipid and energy metabolism disorders in the body. Fecal microbiota transplantation (FMT) can colonize the intestinal tract of obese rats, improving the abundance, diversity and the structure of the flora, activating GPR43 and the downstream mechanisms. This regulation of peptide hormone secretion by endocrine cells can improve metabolic disorders caused by a high-fat diet and may play a significant role in preventing and treating obesity. Biological sciences/Microbiology/Bacteria Health sciences/Endocrinology/Endocrine system and metabolic diseases/Obesity Obesity High fat diet Fecal microbiota transplantation Intestinal flora-SCFAs-GPR43-gastrointestinal peptide Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 1 Introduction Obesity refers to a condition where long-term energy intake exceeds energy expenditure, resulting in excessive energy storage in the form of fat. The accumulation of fat reaches a level that is detrimental to health. It has become one of the major global public health challenges, severely harms human health and quality of life, and is an important independent risk factor for cardiovascular diseases, metabolic syndrome, cancer, and psychological disorders [ 1 – 2 ] . The gut microbiota extensively participates in the host’s fat metabolism and the development of obesity [ 3 ] .Fecal microbiota transplantation (FMT) has gained attention as a novel therapy for modulating the gut microbiota. FMT involves transplanting functional microbial communities isolated from the feces of healthy donors into the gastrointestinal tract of patients via specific routes, which aims to regulate the disordered gut microbiota, restoring the balance in the structure and function of the intestinal microbial community [ 4 ] , which has the potential to become a future therapy for treating obesity and related metabolic disorders [ 5 ] . The gut microbiota exhibits dynamic changes with dietary alterations, and it may be a key factor linking diet and obesity, as it is influenced by diet and responds to dietary changes [ 6 ] . Unhealthy eating habits, such as high-fat and high-sugar diets, can cause dysbiosis of gut microbiota, ultimately leading to obesity and metabolic syndromes like diabetes [ 7 ] . Diet can also affect the gut microbiota and obesity of offspring, with maternal high-fat diets causing gut microbiota imbalances and metabolic disorders that can persist into the offspring’s adulthood [ 8 ] . Diet not only impacts the host’s energy metabolism balance, but also plays a key role in the composition of the gut microbiota [ 9 – 10 ] . The gut microbiota degrades indigestible carbohydrates and dietary fibers in the intestines to produce short-chain fatty acids (SCFAs), such as acetate, propionate, and butyrate [ 11 ] . SCFAs play a key role in chronic metabolic diseases like obesity and diabetes, serving as essential substances for glucose and fatty acid synthesis and metabolism, and maintaining intestinal environmental stability [ 12 – 13 ] . SCFAs activate G protein-coupled receptors (GPCRs) to participate in various cellular physiological metabolic reactions, including regulating the secretion of glucagon-like peptide-1 (GLP-1) and peptide YY (PYY) in the gastrointestinal tract. These gastrointestinal hormones modulate appetite, improve glucose metabolism, and reduce blood sugar levels [ 14 ] . GPR43, a specific receptor for SCFAs of the GPCR family, is widely expressed in the body and is involved in lipid metabolism processes. Gastrin (GAS) and motilin (MTL) are the major gastrointestinal hormones, GAS can promote smooth muscle contraction, accelerate intestinal peristalsis, and promote food digestion, and MTL can promote the contraction of gastric antrum and stomach body, improve the pyloric tension, and delay the gastric emptie [ 15 ] . Based on previous studies, it is hypothesized that a high-fat diet may disrupt the gut microbiota via the “gut microbiota-SCFAs-GPR43-gastrointestinal peptide” pathway. FMT potentially offers protective effects by correcting this pathway in a high-fat diet rat model. Therefore, this study aims to establish an obesity rat model by modifying dietary structure, explore the effects of high-fat diet factors on microbiota, microbiota metabolites, and downstream indicators, and investigate the protective effects and the potential mechanisms of normal microbiota through FMT. 2 Materials and methods 2.1 Animals and diets A total of 160 SPF grade SD rats, male, aged 4 weeks, were provided by Beijing Sibeifu Experimental Animal Technology Co., Ltd. They were housed in the Animal Laboratory of the Institute of Traditional Chinese Medicine, Beijing University of Chinese Medicine, with a temperature of 20–25°C and humidity of 50%-70%. Natural light, good ventilation. A standard rat maintenance pellet feed as normal feed, and a high-fat diet consisting of custom Research Diets-D12492 feed, both were provided by Beijing Sibeifu Biotechnology Co., Ltd. (Animal Ethics Approval Number: BUCM-4-2020082101-3127). 2.2 Main experimental reagents and drugs Anti-GPCR GPR43 (orb159222, Biorbyt), DAB chromogen kit for IHC (Solarbio, Beijing, China), Rat Motilin ELISA Kit, Rat Gastrin ELISA Kit, Rat Glucagon-like Peptide-1 ELISA Kit, Rat Leptin ELISA Kit (Jiangsu Kete Biotechnology Co., Ltd., Nanjing, China), TC and TG assay kits (Xinchuangyuan Biotechnology Co., Ltd., Beijing, China). Neomycin Sulphate and Streptomycin Sulfate were provided by Beijing BioDee Biotechnology Co., Ltd., mixed in a 1:1 ratio, dissolved in saline, and stirred to form a 200 mg/ml antibiotic mixture. 2.3 Experimental models, and drug administration Animals were randomly divided into normal diet group (50 rats) and high-fat diet group (110 rats). They were fed continuously for 10 weeks, with animals in the high-fat diet group that did not meet obesity criteria being excluded, 72 rats were successfully induced obesity. Seven rats from the normal diet group, and 9 rats from the high-fat diet group, were randomly selected to prepare fecal microbiota suspensions. The following experimental groups were established: Normal Control 1 (NC1), Normal Control 2 (NC2), Fecal Microbiota Transplantation 1 (FMT1), receiving normal diet, and two groups within the high-fat diet group, each consisting of 9 rats: Model (M) and Fecal Microbiota Transplantation 2 (FMT2), receiving Research Diets-D12492 high-fat diet. According to the literature [ 16 ] , rats in groups NC2, M, FMT1, and FMT2 were orally gavaged with a mixed solution of Neomycin Sulphate and Streptomycin Sulfate (1:1, 200 mg/kg) for six days, twice daily. Additionally, 6 rats that did not meet obesity criteria were randomly selected from the high-fat diet group for antibiotic intervention. Their intestinal contents were collected one day before antibiotic administration (MB) and one day after antibiotic cessation (MA). After antibiotic-induced pseudo-sterility, fecal microbiota transplantation was performed every other day for six times, i.e., normal feeding was given on days 7, 9, 11, 13, 15, and 17, and on days 8, 10, 12, 14, 16, 18, rats in groups NC1, NC2, and M received saline enema, while rats in groups FMT1 and FMT2 received high-fat microbiota and normal microbiota enema, respectively. 2.4 Observations Animals were assessed for general characteristics, with obesity modeling deemed successful when the body weight of rats in the high-fat diet groups exceeded that of the normal diet groups by 10%. Morphological evaluations of colon and liver tissues were conducted, alongside high-throughput sequencing of intestinal microbiota. Gas chromatography-mass spectrometry (GC-MS) was used to detect short-chain fatty acid expression in intestinal contents. IHC was employed to assess GPR43 expression, while ELISA was used to measure gastrointestinal peptide levels in serum. Furthermore, automated biochemical analyzers were employed to quantify cholesterol and triglyceride levels in serum. 2.5 Statistical methods Statistical analyses were performed using SPSS version 25.0 software. Data were presented as mean ± standard deviation (‾ x ± s). Non-parametric tests were applied when data were not normally distributed. For data normal distribution, one-way analysis of variance (ANOVA) was conducted, followed by the least significant difference (LSD) method to compare between groups. P < 0.05 indicated significant differences. 3 Results 3.1 High-fat diet induced obese rats During the first to sixth weeks of high-fat diet-induced obesity modeling, the body weight of rats in the high-fat diet groups were significantly lower than that of the normal diet groups ( P 0.05). In the ninth and tenth weeks, the body weight of the high-fat diet groups were significantly higher than that of the normal diet groups ( P < 0.05). By the tenth week, 72 out of 110 rats in the high-fat diet groups had an average body weight exceeding that of the normal diet groups by 10%, indicating successful induction of obesity, with a modeling success rate of approximately 65.45%. During the first two weeks, rats in the high-fat diet group exhibited increased water intake, yellowish and disheveled fur, harder and more odorous feces, and prolonged defecation time. In the middle to late stages of modeling, these rats showed signs of slowed movement, reduced activity, poor mental state, decreased fur luster, curling up, hair loss, slightly yellow urine, delayed defecation with sticky feces, and a soiled anus. 3.2 Histological Examination (HE Staining) The structure of colonic tissue of rats in the NC1 group was clear and intact. In the M group, the mucosal epithelial cell arrangement was disordered compared to the NC1 group, with infiltration of inflammatory cells observed, similar conditions were observed in the FMT1 group. The FMT2 group showed improvements compared to the M group. The liver tissue structure in the NC1 group was intact, with neatly arranged hepatocytes and no visible fat droplets. In the M group, hepatocytes were loosely and disorderly arranged with diffusely accumulated fat droplets. Fat droplets were also observed in the FMT1 group. However, the FMT2 group showed a reduction in fat droplets compared to the M group. 3.3 Gut Microbiota The Firmicutes and Bacteroidetes phyla constitute the majority of the gut microbiota in all groups. Examining the Firmicutes/Bacteroidetes ratio, the ratio of M group was 16.76%, the NC1 group was 8.83%, NC2 group was 5.01%, FMT1 group was 28.11%, and FMT2 group was 5.25%. This indicated that a high-fat diet could increase the Firmicutes/Bacteroidetes ratio in rats, transplanting normal gut microbiota could restore the Firmicutes/Bacteroidetes ratio in obese rats to normal levels, while transplanting obese gut microbiota could increase this ratio in normal rats. Additionally, phyla such as Actinobacteria and Proteobacteria, although less dominant, showed significant changes after high-fat diet and fecal microbiota transplantation (FMT) intervention. A high-fat diet increased the structure of these phyla several times, and FMT intervention showed changes similar to those in the Firmicutes/Bacteroidetes ratio. At the class level, the major classes included Clostridia, Bacilli, Actinobacteria, Verrucomicrobia, and Bacteroidia. At the order level, the major orders included Bacteroidales, Clostridiales, Lactobacillales, Erysipelotrichales, Verrucomicrobiales, and Enterobacteriales. At the family level, the major families included Lactobacillaceae, Bacteroidaceae, Erysipelotrichaceae, Eubacteriaceae, Akkermansiaceae, Muribaculaceae, Bacteroidaceae, and Ruminococcaceae. At the genus level, the major genera included Lactobacillus, Allobaculum, Bacteroides, Blautia, Subdoligranulum, Faecalibacterium, and others. Phylogenetic analysis showed that the major phyla in the gut microbiota were Firmicutes, Proteobacteria, Bacteroidetes, and Actinobacteria, with Firmicutes, Proteobacteria, and Bacteroidetes covering the majority of species. Corresponding to the relative abundance results, Lactobacillus, Allobaculum, and Faecalibacterium were the most abundant genera in all five groups. The FMT1 and M groups showed significantly lower abundance of Lactobacillus compared to the NC1, NC2, and FMT2 groups, indicating altered gut environments in these groups, and reflecting gut microbiota dysbiosis induced by a high-fat diet, and FMT can modulate the structure of gut microbiota. Results of diversity analysis showed that the rarefaction curves for all five groups gradually plateaued after 10,000 sequences, indicating that the sample sequencing data were within a reasonable range and further increases in sequencing depth would not significantly affect the number of OTUs. After sampling 20,000 sequences, species abundance was highest in the NC1 group, followed by the FMT2, NC2, FMT1, and M groups. The NC1 group had the highest average species abundance, while the M group had the lowest. All five groups showed a trend of decreasing species abundance and increasing evenness as OTU abundance increased. The characteristics of these curves indicated good species abundance and evenness in the samples, and that the constructed libraries were sufficient to represent the majority of gut bacteria for microbiota diversity analysis, with sequencing depth within a reasonable range. The FMT1 and M groups showed similar dispersion, indicating more similar community composition compared to the NC1, NC2, and FMT2 groups. This demonstrates that different diets and interventions can lead to distinct developments in gut microbial communities. LEfSe results showed significant increases in Gamma-proteobacteria, Proteobacteria, Rhizobiales (family), Atopobiaceae, Akkermansiaceae, Verrucomicrobia (order), Erysipelotrichales (genus), and Allobaculum in the M group compared to the NC1 group. In contrast, Lactobacillaceae (genus), Firmicutes, Blautia, Bacteroidetes (order), Muribaculaceae, Faecalibacterium, and Bacillus were significantly reduced. Compared to the M group, the FMT2 group had increased Lactobacillaceae (genus), Muribaculaceae, Bacteroidetes (order), and Bacillus, while Allobaculum, Erysipelotrichales, Verrucomicrobia (order), Akkermansiaceae, Erysipelotrichales, and Atopobiaceae were significantly reduced. Compared to the M group, the FMT1 group had increased Firmicutes and Clostridia, but decreased Lactobacillales (family, genus). The findings indicated significant differences in gut microbiota between the M group and the NC1 group. There were smaller differences between the M group and the FMT1 group. This suggests that normal microbiota transplantation can help obese rats’ gut microbiota resemble that of normal rats, while obese microbiota transplantation make normal rats’ gut microbiota resemble that of obese rats, showing that FMT could alter the gut microbial structure. 3.4 GPR43 Expression IHC analysis showed that GPR43 was expressed in the liver tissues of all groups of rats, with lower expression levels in the M and FMT1 groups compared to the NC1 and FMT2 groups (Fig. 3). 3.5 Expression of Short-Chain Fatty Acid (SCFA) The results showed that: (1) Acetate: The M and FMT1 groups had significantly lower levels compared to the NC1, NC2, and FMT2 groups ( P < 0.05). (2)Propionate: The FMT1 group had significantly lower levels compared to the NC1 group ( P < 0.05), and both the M and FMT1 groups had significantly lower levels compared to the NC2 and FMT2 groups ( P < 0.05). (3)Butyrate: The M and FMT1 groups had significantly lower levels compared to the NC1, NC2, and FMT2 groups ( P < 0.05). These findings indicated that the levels of SCFAs were reduced in obese rats, suggesting that fecal microbiota transplantation (FMT) may exert effects by influencing the production of SCFAs. Table 1 SCFA Content in Feces of Each Group [M (0.25, 0.75)] Groups n Acetate Propionate Butyrate NC1 9 994.03 (875.26,1176.38) 394.27 (268.30,472.32) 555.52 (466.59,664.57) NC2 9 1447.96 (1149.12,1491.21) 694.19 (442.54,783.40) 451.12 (374.11,577.33) M 9 148.90 (122.11,268.88)* # 102.43 (79.22,134.76) # 96.10 (74.64,134.40)* # FMT1 9 153.76 (147.02,204.65)* # 103.99 (56.472,114.98)* # 100.39 (82.700,113.84)* # FMT2 9 935.71 (623.44,1231.65) ab 501.50 (380.76,678.48) ab 494.26 (378.36,607.54) ab Note: Compared with NC1, * P < 0.05; compared with NC2, # P < 0.05; compared with M, a P < 0.05; compared with FMT1, b P < 0.05. 3.6 Expression of Gastrointestinal Peptide The gastrointestinal tract secreted various regulatory peptides. This study observed the expression levels of PYY, GLP-1, GAS, and MTL in each group: (1) GLP-1: The M and FMT1 groups showed significantly lower levels compared to the NC1 and NC2 groups ( P < 0.05); the FMT2 group had significantly higher levels compared to the M and FMT1 groups ( P 0.05); no significant differences were observed among the NC1, NC2, and FMT2 groups ( P > 0.05). (2)PYY: The M and FMT1 groups had significantly lower levels compared to the NC1 group ( P < 0.05); the FMT2 group had significantly higher levels compared to the M group ( P 0.05). (3)MTL: The M and FMT1 groups showed significantly higher levels compared to the NC1, NC2, and FMT2 groups ( P 0.05); no significant differences were observed among the NC1, NC2, and FMT2 groups ( P > 0.05). (4) GAS: No significant differences were observed among the groups ( P > 0.05). Table 2 Gastrointestinal Peptide Levels in Each Group of Rats (‾ x ± s) Groups n GLP-1 PYY MTL GAS NC1 6 3.67 ± 0.49 216.88 ± 13.03 336.31 ± 46.84 84.86 ± 16.99 NC2 6 3.67 ± 0.43 192.42 ± 28.84 331.74 ± 34.04 92.90 ± 14.07 M 6 2.49 ± 0.51* # 169.85 ± 11.55* 453.17 ± 24.58* # 84.58 ± 11.59 FMT1 6 2.93 ± 0.30* # 179.44 ± 36.56* 415.39 ± 77.30* # 88.37 ± 3.33 FMT2 6 3.88 ± 0.11 ab 205.31 ± 17.07 a 325.93 ± 41.37 ab 82.94 ± 6.60 Note: Compared with NC1, * P < 0.05; compared with NC2, # P < 0.05; compared with M, a P < 0.05; compared with FMT1, b P < 0.05. 3.7 Serum TC and TG Levels Long-term high-fat diets caused lipid metabolism disorders. Serum TC and TG levels showed that: (1) TC levels: The M, FMT1, and FMT2 groups had significantly higher serum TC levels compared to the NC1 and NC2 groups ( P < 0.05); the FMT1 and FMT2 groups had significantly lower TC levels compared to the M group ( P 0.05). (2) TG levels: The NC2, FMT1, and FMT2 groups had significantly lower serum TG levels compared to the NC1 group ( P 0.05); the FMT1 group had significantly lower TG levels compared to the M group ( P 0.05). Table 3 TC and TG Levels in Each Group of Rats (‾ x ± s,mmol/L) Groups n TC TG NC1 6 1.34 ± 0.22 1.06 ± 0.28 NC2 6 1.11 ± 0.17 0.70 ± 0.18* M 6 2.26 ± 0.39* # 0.89 ± 0.23 FMT1 6 1.69 ± 0.14* #a 0.61 ± 0.15* a FMT2 6 1.67 ± 0.17* #a 0.65 ± 0.12* Note: Compared with NC1, * P < 0.05; compared with NC2, # P < 0.05; compared with M, a P < 0.05. 4 Discussion 4.1 The Impact of FMT on Gut Microbiota in Obese Rats Gut microbiota is closely related to energy balance and metabolism of its host. Obesity is associated with changes in the relative abundance of the two main phyla in the gut microbiota, Bacteroidetes and Firmicutes [ 17 ] . The tendency toward obesity or increased body fat may be determined by the ratio of Bacteroidetes to Firmicutes, which can serve as a biomarker for diagnosing obesity-related metabolic syndrome [ 18 ] . Obesity is one of the indications for FMT in treating dysbiosis-related diseases [ 19 ] . FMT is considered as an effective means of reconstructing gut microbiota. Colonization of beneficial microbiota in the gut of obese rats can improve the gut environment and correct dysbiosis [ 20 ] . In this study, a high-fat diet increased the Firmicutes/Bacteroidetes ratio in rats. Normal fecal microbiota transplantation restored this ratio in obese rats to that of normal rats, while fecal microbiota transplantation from obese rats increased this ratio in normal rats. The abundance and diversity of gut microbiota in the M and FMT1 groups were lower than those in the NC1 group, suggesting that a high-fat diet alters the abundance and diversity of gut microbiota. Overall, the structure of gut microbiota of the M and FMT1 groups was more similar, and was different from the NC1 group, indicating that transplantation of gut microbiota from obese rats could disrupt the gut microbiota of normal rats to some extent. Previous studies have found that Clostridium, a bacterium that can reduce fat content, increases in abundance to reduce fat content to some extent. If Clostridium abundance decreases, serum leptin levels increase. Supplementing probiotics to increase Clostridium abundance can enhance host sensitivity to leptin. The Ruminococcaceae family is also closely related to obesity, with its abundance significantly increased in the gut of high-fat diet-induced obese mice [ 21 ] , which was consistent with the results in this study. Akkermansia muciniphila, a Gram-negative bacterium, its colonization is generally believed to be negatively correlated with waist circumference, body weight, and body fat. It can reduce fat accumulation, enhance metabolism, and promote weight loss, with lower abundance in obese individuals [ 22 ] . However, in this study, Akkermansia muciniphila significantly increased in the high-fat groups, possibly due to the self-regulatory response of rats to a high-fat diet or the use of antibiotics. After neomycin-containing antibiotic intervention, Verrucomicrobia significantly increased in mice [ 23 ] . Oral vancomycin reduced nutrient absorption and increased Akkermansia muciniphila by regulating intestinal barrier function [ 24 ] . Streptomycin and vancomycin used in this study are narrow-spectrum antibiotics, and Akkermansia muciniphila is a dominant genus in the Verrucomicrobia phylum. Therefore, the increase in Akkermansia muciniphila may be related to antibiotic use. 4.2 Normal Fecal Microbiota Exerts Effects through the SCFAs-GPR43-Gastrointestinal Peptide Pathway One of the main ways to produce energy in the colon is through the metabolism of SCFAs, which increases lipid metabolic rates [ 25 ] . SCFAs not only serve as substrates for lipid and energy metabolism, but also act as regulatory factors to modulate host physiological metabolism [ 26 ] . An improper diet is the main cause of obesity and metabolic syndrome. When the diet changes, gut microbiota rapidly respond to these changes [ 27 ] . Diet can affect SCFA levels by altering the type and abundance of gut microbiota, thereby influencing obesity. SCFAs play a crucial role in maintaining gut health and function, providing energy and nutrients necessary for bacterial growth and reproduction. They maintain normal osmotic pressure, epithelial cell integrity, and promote epithelial cell repair in the gut [ 28 ] . SCFAs can exert various effects to maintain and improve gut health, control blood glucose homeostasis by regulating systemic energy balance, and have the potential to alleviate obesity and diabetes [ 29 ] . The results indicated that SCFA levels were significantly reduced in the M group, suggesting that microbiota transplantation exerts effects by influencing SCFA production. In addition to energy metabolism, SCFAs can act as signaling molecules, binding to G-protein-coupled receptors (GPCRs), stimulating GLP-1 secretion and PYY production, inhibiting gut motility, controlling appetite, altering transit time, and affecting lipid accumulation [ 30 ] . GPR43 is closely related to gut function and lipid metabolism, controlling the body’s energy utilization rate and maintaining a stable metabolic environment. Mice lacking GPR43 become obese under normal dietary conditions, while overexpression of GPR43 in mice prevents obesity even with a high-fat diet [ 31 ] . Studies indicate that a high-fat diet affects metabolism by reducing GPR43 expression. GPR43 can co-express with GLP-1, mediating SCFA-induced GLP-1 secretion. The fat-reducing effect of GLP-1 is widely studied and recognized, and GLP-1 levels are reduced in patients with obesity and metabolic syndrome [ 32 ] . As an incretin, GLP-1 plays a key role in regulating blood glucose levels and insulin secretion. GLP-1 agonists have been shown to reduce appetite, decrease energy intake, delay gastric emptying, and subsequently reduce body weight [ 33 ] , and also impact cardiometabolic risk factors, reducing the risk of cardiovascular events associated with obesity [ 34 – 35 ] . Increased PYY production can reduce gut motility, increasing energy acquisition post-meal and enhancing satiety [ 36 ] . The results indicate that GLP-1 levels and PYY expression were lower in the M and FMT1 groups compared to the NC1 group. Reduced MTL can slow digestion and absorption, reducing postprandial hyperglycemia and hyperlipidemia [ 37 ] . Reduced motilin promotes gastric emptying, playing an important role in regulating gastrointestinal motility [ 38 ] . Although there were no significant differences in GAS levels among the groups, the secretion volume may not change significantly, and the response may be enhanced. The MTL levels in the M and FMT1 groups were higher than in the NC1 group, suggesting MTL activation in obese rats, increasing hunger by acting on the feeding center, and subsequently increasing food intake. Chen Luman et al. found through animal experiments that activation of the MTL receptor increases appetite and body weight of rats [ 39 ] . Reports indicate that MTL levels are higher in obese patients than in healthy individuals [ 40 ] . Stabilizing gastrointestinal hormone levels helps improve gastrointestinal function, consistent with the MTL expression results of this study. 4.3 Pathology Conditions and Changes in Serum TC and TG In M group, intestinal mucosal epithelial cells were disordered, with visible inflammatory cell infiltration, suggesting that a high-fat diet affects normal gastrointestinal digestion and excretion functions, leading to abnormal and pathological changes. Hepatocytes in M group were loosely and disorderedly arranged, with diffuse accumulation of lipid droplets, indicating that obese rats exhibited fatty liver and increased intra-abdominal fat. Due to the gut-liver axis, the liver is directly exposed to gut microbiota and their products. When gut microbiota is imbalanced, the permeability and integrity of the intestinal wall are affected, allowing a large number of bacterial products to enter the liver, leading to liver dysfunction [ 41 ] . An intact intestinal mucosal barrier and liver defense function are crucial for maintaining internal homeostasis. Numerous studies have shown that once the balance between the liver and the intestine is disrupted, it will lead to pathological changes such as hepatic steatosis and hepatitis [ 42 ] . A retrospective cohort study indicated that obesity is a significant risk factor for fatty liver disease, as the liver is a key site for lipid metabolism [ 43 ] . SCFAs can maintain the integrity of the intestinal barrier, preventing intestinal toxins like lipopolysaccharides from being absorbed through the intestinal epithelium and invading the liver. In colon epithelial cells, SCFAs bind to GPR43, activate caspase-1, and release interleukin-18, promoting intestinal epithelial repair [ 44 ] . Obesity damages the intestinal barrier, accelerating obesity, while SCFAs help improve the intestinal barrier [ 45 ] . Obese individuals have high visceral fat content, leading to the release of large amounts of free fatty acids into the liver, promoting TG synthesis. Excessive fat deposition can alter lipase activity, subsequently increasing TC levels [ 46 ] . Obese patients often exhibit altered blood lipid levels, typically showing elevated TG and TC levels. A high-fat diet increases fat deposition in rats, potentially accompanied by an overall increase in metabolic levels. On one hand, the intake of a high-fat diet requires the production of large amounts of bile acids for digestion and absorption, promoting the liver to accelerate TC breakdown to form bile acids. On the other hand, a high-fat diet can increase liver enzyme activity, speeding up TC synthesis. Based on the results of this study, it is speculated that the total metabolic level of TC in high-fat diet rats tends to increase, with the efficiency of TC synthesis exceeding that of breakdown, resulting in significant changes in serum TC levels in the FMT1 group and M group compared to the NC1 group. The TG levels in the FMT1 group and M group were lower than those in the NC1 group, possibly due to fasting for 12 hours before sampling, causing abnormal elevation of indicators in the NC1 group due to hunger, thus interfering with serum TG levels and causing inconsistent results compared to previous findings. The results of this study were consistent with revious reports [ 47 ] . Additionally, the TG levels in the antibiotic intervened NC2 group were lower than those in the NC1 group, suggesting that antibiotic intervention may also be related to the atypical TG levels observed in this study. 5 Conclusions In summary, a high-fat diet could induce related pathological reactions by affecting the gut microbiota-SCFAs-GPR43-gastrointestinal peptide pathway, lead to gut microbiota dysbiosis, SCFA metabolism disorders, and subsequent activation of GPR43 to release PYY, GLP-1, GAS, and MTL, result in lipid and energy metabolism disorders in the body. Normal microbiota can be colonized in the gut of obese rats through fecal microbiota transplantation, improving reduced abundance, diversity, and disordered microbiota structure, activating GPR43, initiating downstream mechanisms, regulating the secretion of peptide hormones by endocrine cells, and alleviating metabolic disorders induced by a high-fat diet. This suggests that gut microbiota can be a new target for treating obesity and other metabolic diseases, promoting a positive cycle of body metabolism. The SCFAs in this study included a combination of acetate, propionate, and butyrate, but their individual mechanisms in regulating obesity remain unclear. The specific pathways of different SCFAs’ mechanisms of action, the synergy or contradictions of the same SCFA in different signaling pathways, and the synergy or contradictions of different SCFAs in the same signaling pathway could be explored in future studies. Declarations Funding This study was supported by the the Fundamental Research Funds for the Central Universities for the Unveiling of the List of Commander-in-Chief Programs in Beijing University of Chinese Medicine（2024-JYB-JBZD-038）; National Natural Science Foundation of China (81704121); Research on the Four Famous Doctors in Beijing and their Main Inheritors, and the Inheritance of the Experiences of Famous Doctors of Yanjing（BUCM-2023-JS-FW-077）; High-level traditional Chinese medicine key subjects construction project of National Administration of Traditional Chinese Medicine—Beijing University of Chinese Medicine,Chinese Medicine Epidemic Disease（No.zyyzdxk-2023264） Author Contributions He Yu, Xiaomei Wang, and Xu Wang designed this study; Lijun Cui, Shaoyang Liu, Chen Bai, Jianhua Zhen,and Tiegang Liu have undergone the experiment; Xiaohong Gu, Xiaomei Wang, and Xu Wang provided experimental guidance; Lijun Cui, Xiaomei Wang, Xu Wang, and He Yu wrote a manuscript; HeYu and Xiaohong Gu funded this study. 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Role of Glucagon-Like Peptide-1 Receptor Agonists in Achieving Weight Loss and Improving Cardiovascular Outcomes in People With Overweight and Obesity.J Am Heart Assoc. 2023,12(11):e029282. doi: 10.1161/JAHA.122.029282 . Schroeder BO, Bäckhed F. Signals from the gut microbiota to distant organs in physiology and disease. Nat Med,2016, 22(10):1079–1089. Zhang SS, Tian ZL, Feng S, et al. Role of motilin after duodenal-jejunal bypass in the improvement of random blood glucose in type 2 diabetes rat model. Chinese Journal of Gastroenterology and Hepatology, 2019, 28(10): 1140–1143. Gao P, Tu JH. Effects of Modified Shenling Baizhu Decoction on obese type 2 diabetes with spleen deficiency and dampness syndrome: observations on the regulation of adipokines and gastrointestinal hormones in patients. Electronic Journal of Practical Gynecological Endocrinology, 2020, 7(23): 164–165. doi: 10.16484/j.cnki.issn2095-8803.2020.23.105 . Chen LM, Duan XY, Guan ZM. 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Zhang S, Zhao J, Xie F, et al.Dietary fiber-derived short-chain fatty acids: A potential therapeutic target to alleviate obesity-related nonalcoholic fatty liver disease.Obes Rev. 2021,22(11):e13316. doi: 10.1111/obr.13316 . Diao H, Jiao AR, Yu B, et al.Gastric infusion of short-chain fatty acids can improve intestinal barrier function in weaned piglets.Genes Nutr. 2019,14:4. doi: 10.1186/s12263-019-0626-x . Bai Y, Bao XL, Zhao DD, et al. Mechanism study of jiang Tang San Hao Formula improving glucolipid metabolism in diet-induced obese mice. China Journal of Traditional Chinese Medicine and Pharmacy, 2020, 35(5): 2253–2258. Brower Marcia, Grace Martha, Kotz Catherine M, et al. Comparative analysis of growth characteristics of Sprague Dawley rats obtained from different sources.. Laboratory animal research,2015,31(4):166–173. 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Liver Tissues of Each Group\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-6307329/v1/0132bdcbf7817bc825cc2600.png"},{"id":81267308,"identity":"e5be7f56-6d8b-4602-970c-0b5745c037b7","added_by":"auto","created_at":"2025-04-24 07:41:14","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":546263,"visible":true,"origin":"","legend":"\u003cp\u003eGPR43 Expression in Liver Tissues of Each Group\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-6307329/v1/6e416192e832c898674b1c2d.png"},{"id":81267262,"identity":"1dbfb28b-e4a7-4dba-854d-e9914a766060","added_by":"auto","created_at":"2025-04-24 07:41:12","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":139829,"visible":true,"origin":"","legend":"\u003cp\u003eUnnumbered image in the Results section.\u003c/p\u003e","description":"","filename":"Uf1.png","url":"https://assets-eu.researchsquare.com/files/rs-6307329/v1/113f0f7088b0c392f15f1947.png"},{"id":81267255,"identity":"6bb8d07b-4f0f-4194-a4e7-136b7d98a5c1","added_by":"auto","created_at":"2025-04-24 07:41:12","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":76351,"visible":true,"origin":"","legend":"\u003cp\u003eUnnumbered image in the Results section.\u003c/p\u003e","description":"","filename":"Uf2.png","url":"https://assets-eu.researchsquare.com/files/rs-6307329/v1/62763e68636dac4d3fd4d99a.png"},{"id":90516065,"identity":"3ebe6bb8-d8e7-4015-835e-c4782d00d463","added_by":"auto","created_at":"2025-09-03 14:35:20","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":2493024,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-6307329/v1/21db4200-dd4a-4ccc-8194-4e701ecf18b4.pdf"},{"id":81267268,"identity":"a5f0c5c5-e947-43f7-bb52-e091f3ed0ee8","added_by":"auto","created_at":"2025-04-24 07:41:12","extension":"docx","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":10617006,"visible":true,"origin":"","legend":"","description":"","filename":"Data.docx","url":"https://assets-eu.researchsquare.com/files/rs-6307329/v1/6436f6d9ac4f574b0b0e6d19.docx"}],"financialInterests":"There is \u003cb\u003eNO\u003c/b\u003e conflict of interest to disclose","formattedTitle":"Effect of fecal microbiota transplantation on “intestinal flora-SCFAs-GPR43 - gastrointestinal peptide” pathway in rats with high-fat diet","fulltext":[{"header":"1 Introduction","content":"\u003cp\u003eObesity refers to a condition where long-term energy intake exceeds energy expenditure, resulting in excessive energy storage in the form of fat. The accumulation of fat reaches a level that is detrimental to health. It has become one of the major global public health challenges, severely harms human health and quality of life, and is an important independent risk factor for cardiovascular diseases, metabolic syndrome, cancer, and psychological disorders\u003csup\u003e[\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e]\u003c/sup\u003e. The gut microbiota extensively participates in the host\u0026rsquo;s fat metabolism and the development of obesity\u003csup\u003e[\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e]\u003c/sup\u003e.Fecal microbiota transplantation (FMT) has gained attention as a novel therapy for modulating the gut microbiota. FMT involves transplanting functional microbial communities isolated from the feces of healthy donors into the gastrointestinal tract of patients via specific routes, which aims to regulate the disordered gut microbiota, restoring the balance in the structure and function of the intestinal microbial community\u003csup\u003e[\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e]\u003c/sup\u003e, which has the potential to become a future therapy for treating obesity and related metabolic disorders\u003csup\u003e[\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e]\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eThe gut microbiota exhibits dynamic changes with dietary alterations, and it may be a key factor linking diet and obesity, as it is influenced by diet and responds to dietary changes\u003csup\u003e[\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]\u003c/sup\u003e. Unhealthy eating habits, such as high-fat and high-sugar diets, can cause dysbiosis of gut microbiota, ultimately leading to obesity and metabolic syndromes like diabetes\u003csup\u003e[\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e]\u003c/sup\u003e. Diet can also affect the gut microbiota and obesity of offspring, with maternal high-fat diets causing gut microbiota imbalances and metabolic disorders that can persist into the offspring\u0026rsquo;s adulthood\u003csup\u003e[\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e]\u003c/sup\u003e. Diet not only impacts the host\u0026rsquo;s energy metabolism balance, but also plays a key role in the composition of the gut microbiota\u003csup\u003e[\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e]\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eThe gut microbiota degrades indigestible carbohydrates and dietary fibers in the intestines to produce short-chain fatty acids (SCFAs), such as acetate, propionate, and butyrate\u003csup\u003e[\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e]\u003c/sup\u003e. SCFAs play a key role in chronic metabolic diseases like obesity and diabetes, serving as essential substances for glucose and fatty acid synthesis and metabolism, and maintaining intestinal environmental stability\u003csup\u003e[\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e]\u003c/sup\u003e. SCFAs activate G protein-coupled receptors (GPCRs) to participate in various cellular physiological metabolic reactions, including regulating the secretion of glucagon-like peptide-1 (GLP-1) and peptide YY (PYY) in the gastrointestinal tract. These gastrointestinal hormones modulate appetite, improve glucose metabolism, and reduce blood sugar levels \u003csup\u003e[\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e]\u003c/sup\u003e. GPR43, a specific receptor for SCFAs of the GPCR family, is widely expressed in the body and is involved in lipid metabolism processes. Gastrin (GAS) and motilin (MTL) are the major gastrointestinal hormones, GAS can promote smooth muscle contraction, accelerate intestinal peristalsis, and promote food digestion, and MTL can promote the contraction of gastric antrum and stomach body, improve the pyloric tension, and delay the gastric emptie\u003csup\u003e[\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e]\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eBased on previous studies, it is hypothesized that a high-fat diet may disrupt the gut microbiota via the \u0026ldquo;gut microbiota-SCFAs-GPR43-gastrointestinal peptide\u0026rdquo; pathway. FMT potentially offers protective effects by correcting this pathway in a high-fat diet rat model. Therefore, this study aims to establish an obesity rat model by modifying dietary structure, explore the effects of high-fat diet factors on microbiota, microbiota metabolites, and downstream indicators, and investigate the protective effects and the potential mechanisms of normal microbiota through FMT.\u003c/p\u003e"},{"header":"2 Materials and methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003e2.1 Animals and diets\u003c/h2\u003e \u003cp\u003eA total of 160 SPF grade SD rats, male, aged 4 weeks, were provided by Beijing Sibeifu Experimental Animal Technology Co., Ltd. They were housed in the Animal Laboratory of the Institute of Traditional Chinese Medicine, Beijing University of Chinese Medicine, with a temperature of 20\u0026ndash;25\u0026deg;C and humidity of 50%-70%. Natural light, good ventilation. A standard rat maintenance pellet feed as normal feed, and a high-fat diet consisting of custom Research Diets-D12492 feed, both were provided by Beijing Sibeifu Biotechnology Co., Ltd. (Animal Ethics Approval Number: BUCM-4-2020082101-3127).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003e2.2 Main experimental reagents and drugs\u003c/h2\u003e \u003cp\u003eAnti-GPCR GPR43 (orb159222, Biorbyt), DAB chromogen kit for IHC (Solarbio, Beijing, China), Rat Motilin ELISA Kit, Rat Gastrin ELISA Kit, Rat Glucagon-like Peptide-1 ELISA Kit, Rat Leptin ELISA Kit (Jiangsu Kete Biotechnology Co., Ltd., Nanjing, China), TC and TG assay kits (Xinchuangyuan Biotechnology Co., Ltd., Beijing, China). Neomycin Sulphate and Streptomycin Sulfate were provided by Beijing BioDee Biotechnology Co., Ltd., mixed in a 1:1 ratio, dissolved in saline, and stirred to form a 200 mg/ml antibiotic mixture.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003e2.3 Experimental models, and drug administration\u003c/h2\u003e \u003cp\u003eAnimals were randomly divided into normal diet group (50 rats) and high-fat diet group (110 rats). They were fed continuously for 10 weeks, with animals in the high-fat diet group that did not meet obesity criteria being excluded, 72 rats were successfully induced obesity. Seven rats from the normal diet group, and 9 rats from the high-fat diet group, were randomly selected to prepare fecal microbiota suspensions. The following experimental groups were established: Normal Control 1 (NC1), Normal Control 2 (NC2), Fecal Microbiota Transplantation 1 (FMT1), receiving normal diet, and two groups within the high-fat diet group, each consisting of 9 rats: Model (M) and Fecal Microbiota Transplantation 2 (FMT2), receiving Research Diets-D12492 high-fat diet.\u003c/p\u003e \u003cp\u003eAccording to the literature\u003csup\u003e[\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e]\u003c/sup\u003e, rats in groups NC2, M, FMT1, and FMT2 were orally gavaged with a mixed solution of Neomycin Sulphate and Streptomycin Sulfate (1:1, 200 mg/kg) for six days, twice daily. Additionally, 6 rats that did not meet obesity criteria were randomly selected from the high-fat diet group for antibiotic intervention. Their intestinal contents were collected one day before antibiotic administration (MB) and one day after antibiotic cessation (MA). After antibiotic-induced pseudo-sterility, fecal microbiota transplantation was performed every other day for six times, i.e., normal feeding was given on days 7, 9, 11, 13, 15, and 17, and on days 8, 10, 12, 14, 16, 18, rats in groups NC1, NC2, and M received saline enema, while rats in groups FMT1 and FMT2 received high-fat microbiota and normal microbiota enema, respectively.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec6\" class=\"Section2\"\u003e \u003ch2\u003e2.4 Observations\u003c/h2\u003e \u003cp\u003eAnimals were assessed for general characteristics, with obesity modeling deemed successful when the body weight of rats in the high-fat diet groups exceeded that of the normal diet groups by 10%. Morphological evaluations of colon and liver tissues were conducted, alongside high-throughput sequencing of intestinal microbiota. Gas chromatography-mass spectrometry (GC-MS) was used to detect short-chain fatty acid expression in intestinal contents. IHC was employed to assess GPR43 expression, while ELISA was used to measure gastrointestinal peptide levels in serum. Furthermore, automated biochemical analyzers were employed to quantify cholesterol and triglyceride levels in serum.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec7\" class=\"Section2\"\u003e \u003ch2\u003e2.5 Statistical methods\u003c/h2\u003e \u003cp\u003eStatistical analyses were performed using SPSS version 25.0 software. Data were presented as mean\u0026thinsp;\u0026plusmn;\u0026thinsp;standard deviation (\u0026oline;\u003cem\u003ex\u003c/em\u003e\u0026thinsp;\u0026plusmn;\u0026thinsp;s). Non-parametric tests were applied when data were not normally distributed. For data normal distribution, one-way analysis of variance (ANOVA) was conducted, followed by the least significant difference (LSD) method to compare between groups. \u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05 indicated significant differences.\u003c/p\u003e \u003c/div\u003e"},{"header":"3 Results","content":"\u003cdiv id=\"Sec9\" class=\"Section2\"\u003e \u003ch2\u003e3.1 High-fat diet induced obese rats\u003c/h2\u003e \u003cp\u003eDuring the first to sixth weeks of high-fat diet-induced obesity modeling, the body weight of rats in the high-fat diet groups were significantly lower than that of the normal diet groups (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05). In the seventh and eighth weeks, there was no significant difference in body weight between the high-fat and normal diet groups (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026gt;\u0026thinsp;0.05). In the ninth and tenth weeks, the body weight of the high-fat diet groups were significantly higher than that of the normal diet groups (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05). By the tenth week, 72 out of 110 rats in the high-fat diet groups had an average body weight exceeding that of the normal diet groups by 10%, indicating successful induction of obesity, with a modeling success rate of approximately 65.45%.\u003c/p\u003e \u003cp\u003eDuring the first two weeks, rats in the high-fat diet group exhibited increased water intake, yellowish and disheveled fur, harder and more odorous feces, and prolonged defecation time. In the middle to late stages of modeling, these rats showed signs of slowed movement, reduced activity, poor mental state, decreased fur luster, curling up, hair loss, slightly yellow urine, delayed defecation with sticky feces, and a soiled anus.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec10\" class=\"Section2\"\u003e \u003ch2\u003e3.2 Histological Examination (HE Staining)\u003c/h2\u003e \u003cp\u003eThe structure of colonic tissue of rats in the NC1 group was clear and intact. In the M group, the mucosal epithelial cell arrangement was disordered compared to the NC1 group, with infiltration of inflammatory cells observed, similar conditions were observed in the FMT1 group. The FMT2 group showed improvements compared to the M group. The liver tissue structure in the NC1 group was intact, with neatly arranged hepatocytes and no visible fat droplets. In the M group, hepatocytes were loosely and disorderly arranged with diffusely accumulated fat droplets. Fat droplets were also observed in the FMT1 group. However, the FMT2 group showed a reduction in fat droplets compared to the M group.\u003c/p\u003e \u003cdiv id=\"Sec11\" class=\"Section2\"\u003e \u003ch2\u003e3.3 Gut Microbiota\u003c/h2\u003e \u003cp\u003eThe Firmicutes and Bacteroidetes phyla constitute the majority of the gut microbiota in all groups. Examining the Firmicutes/Bacteroidetes ratio, the ratio of M group was 16.76%, the NC1 group was 8.83%, NC2 group was 5.01%, FMT1 group was 28.11%, and FMT2 group was 5.25%. This indicated that a high-fat diet could increase the Firmicutes/Bacteroidetes ratio in rats, transplanting normal gut microbiota could restore the Firmicutes/Bacteroidetes ratio in obese rats to normal levels, while transplanting obese gut microbiota could increase this ratio in normal rats.\u003c/p\u003e \u003cp\u003eAdditionally, phyla such as Actinobacteria and Proteobacteria, although less dominant, showed significant changes after high-fat diet and fecal microbiota transplantation (FMT) intervention. A high-fat diet increased the structure of these phyla several times, and FMT intervention showed changes similar to those in the Firmicutes/Bacteroidetes ratio. At the class level, the major classes included Clostridia, Bacilli, Actinobacteria, Verrucomicrobia, and Bacteroidia. At the order level, the major orders included Bacteroidales, Clostridiales, Lactobacillales, Erysipelotrichales, Verrucomicrobiales, and Enterobacteriales. At the family level, the major families included Lactobacillaceae, Bacteroidaceae, Erysipelotrichaceae, Eubacteriaceae, Akkermansiaceae, Muribaculaceae, Bacteroidaceae, and Ruminococcaceae. At the genus level, the major genera included Lactobacillus, Allobaculum, Bacteroides, Blautia, Subdoligranulum, Faecalibacterium, and others.\u003c/p\u003e \u003cp\u003ePhylogenetic analysis showed that the major phyla in the gut microbiota were Firmicutes, Proteobacteria, Bacteroidetes, and Actinobacteria, with Firmicutes, Proteobacteria, and Bacteroidetes covering the majority of species. Corresponding to the relative abundance results, Lactobacillus, Allobaculum, and Faecalibacterium were the most abundant genera in all five groups. The FMT1 and M groups showed significantly lower abundance of Lactobacillus compared to the NC1, NC2, and FMT2 groups, indicating altered gut environments in these groups, and reflecting gut microbiota dysbiosis induced by a high-fat diet, and FMT can modulate the structure of gut microbiota.\u003c/p\u003e \u003cp\u003eResults of diversity analysis showed that the rarefaction curves for all five groups gradually plateaued after 10,000 sequences, indicating that the sample sequencing data were within a reasonable range and further increases in sequencing depth would not significantly affect the number of OTUs. After sampling 20,000 sequences, species abundance was highest in the NC1 group, followed by the FMT2, NC2, FMT1, and M groups. The NC1 group had the highest average species abundance, while the M group had the lowest. All five groups showed a trend of decreasing species abundance and increasing evenness as OTU abundance increased.\u003c/p\u003e \u003cp\u003eThe characteristics of these curves indicated good species abundance and evenness in the samples, and that the constructed libraries were sufficient to represent the majority of gut bacteria for microbiota diversity analysis, with sequencing depth within a reasonable range. The FMT1 and M groups showed similar dispersion, indicating more similar community composition compared to the NC1, NC2, and FMT2 groups. This demonstrates that different diets and interventions can lead to distinct developments in gut microbial communities.\u003c/p\u003e \u003cp\u003eLEfSe results showed significant increases in Gamma-proteobacteria, Proteobacteria, Rhizobiales (family), Atopobiaceae, Akkermansiaceae, Verrucomicrobia (order), Erysipelotrichales (genus), and Allobaculum in the M group compared to the NC1 group. In contrast, Lactobacillaceae (genus), Firmicutes, Blautia, Bacteroidetes (order), Muribaculaceae, Faecalibacterium, and Bacillus were significantly reduced. Compared to the M group, the FMT2 group had increased Lactobacillaceae (genus), Muribaculaceae, Bacteroidetes (order), and Bacillus, while Allobaculum, Erysipelotrichales, Verrucomicrobia (order), Akkermansiaceae, Erysipelotrichales, and Atopobiaceae were significantly reduced. Compared to the M group, the FMT1 group had increased Firmicutes and Clostridia, but decreased Lactobacillales (family, genus).\u003c/p\u003e \u003cp\u003eThe findings indicated significant differences in gut microbiota between the M group and the NC1 group. There were smaller differences between the M group and the FMT1 group. This suggests that normal microbiota transplantation can help obese rats\u0026rsquo; gut microbiota resemble that of normal rats, while obese microbiota transplantation make normal rats\u0026rsquo; gut microbiota resemble that of obese rats, showing that FMT could alter the gut microbial structure.\u003c/p\u003e \u003cdiv id=\"Sec12\" class=\"Section2\"\u003e \u003ch2\u003e3.4 GPR43 Expression\u003c/h2\u003e \u003cp\u003eIHC analysis showed that GPR43 was expressed in the liver tissues of all groups of rats, with lower expression levels in the M and FMT1 groups compared to the NC1 and FMT2 groups (Fig.\u0026nbsp;3).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec13\" class=\"Section2\"\u003e \u003ch2\u003e3.5 Expression of Short-Chain Fatty Acid (SCFA)\u003c/h2\u003e \u003cp\u003eThe results showed that: (1) Acetate: The M and FMT1 groups had significantly lower levels compared to the NC1, NC2, and FMT2 groups (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05). (2)Propionate: The FMT1 group had significantly lower levels compared to the NC1 group (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05), and both the M and FMT1 groups had significantly lower levels compared to the NC2 and FMT2 groups (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05). (3)Butyrate: The M and FMT1 groups had significantly lower levels compared to the NC1, NC2, and FMT2 groups (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05). These findings indicated that the levels of SCFAs were reduced in obese rats, suggesting that fecal microbiota transplantation (FMT) may exert effects by influencing the production of SCFAs.\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\u003eSCFA Content in Feces of Each Group [M (0.25, 0.75)]\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"5\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" 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 \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGroups\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003en\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eAcetate\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003ePropionate\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eButyrate\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNC1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e994.03\u003c/p\u003e \u003cp\u003e(875.26,1176.38)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e394.27\u003c/p\u003e \u003cp\u003e(268.30,472.32)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e555.52\u003c/p\u003e \u003cp\u003e(466.59,664.57)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNC2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1447.96\u003c/p\u003e \u003cp\u003e(1149.12,1491.21)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e694.19\u003c/p\u003e \u003cp\u003e(442.54,783.40)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e451.12\u003c/p\u003e \u003cp\u003e(374.11,577.33)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eM\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e148.90\u003c/p\u003e \u003cp\u003e(122.11,268.88)*\u003csup\u003e#\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e102.43\u003c/p\u003e \u003cp\u003e(79.22,134.76)\u003csup\u003e#\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e96.10\u003c/p\u003e \u003cp\u003e(74.64,134.40)*\u003csup\u003e#\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eFMT1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e153.76\u003c/p\u003e \u003cp\u003e(147.02,204.65)*\u003csup\u003e#\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e103.99\u003c/p\u003e \u003cp\u003e(56.472,114.98)*\u003csup\u003e#\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e100.39\u003c/p\u003e \u003cp\u003e(82.700,113.84)*\u003csup\u003e#\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eFMT2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e935.71\u003c/p\u003e \u003cp\u003e(623.44,1231.65)\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e501.50\u003c/p\u003e \u003cp\u003e(380.76,678.48)\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e494.26\u003c/p\u003e \u003cp\u003e(378.36,607.54)\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"5\"\u003eNote: Compared with NC1, *\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05; compared with NC2, \u003csup\u003e#\u003c/sup\u003e\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05; compared with M, \u003csup\u003ea\u003c/sup\u003e\u003cem\u003eP\u003c/em\u003e \u0026lt; 0.05; compared with FMT1, \u003csup\u003eb\u003c/sup\u003e\u003cem\u003eP\u003c/em\u003e \u0026lt; 0.05.\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec14\" class=\"Section2\"\u003e \u003ch2\u003e3.6 Expression of Gastrointestinal Peptide\u003c/h2\u003e \u003cp\u003eThe gastrointestinal tract secreted various regulatory peptides. This study observed the expression levels of PYY, GLP-1, GAS, and MTL in each group: (1) GLP-1: The M and FMT1 groups showed significantly lower levels compared to the NC1 and NC2 groups (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05); the FMT2 group had significantly higher levels compared to the M and FMT1 groups (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05); there was no significant difference between the M and FMT1 groups (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026gt;\u0026thinsp;0.05); no significant differences were observed among the NC1, NC2, and FMT2 groups (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026gt;\u0026thinsp;0.05).\u003c/p\u003e \u003cp\u003e(2)PYY: The M and FMT1 groups had significantly lower levels compared to the NC1 group (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05); the FMT2 group had significantly higher levels compared to the M group (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05); no significant differences were observed among the other groups (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026gt;\u0026thinsp;0.05). (3)MTL: The M and FMT1 groups showed significantly higher levels compared to the NC1, NC2, and FMT2 groups (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05); there was no significant difference between the M and FMT1 groups (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026gt;\u0026thinsp;0.05); no significant differences were observed among the NC1, NC2, and FMT2 groups (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026gt;\u0026thinsp;0.05). (4) GAS: No significant differences were observed among the groups (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026gt;\u0026thinsp;0.05).\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\u003eGastrointestinal Peptide Levels in Each Group of Rats (\u0026oline;\u003cem\u003ex\u003c/em\u003e\u0026thinsp;\u0026plusmn;\u0026thinsp;s)\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"6\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" 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=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGroups\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003en\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eGLP-1\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003ePYY\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eMTL\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eGAS\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNC1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e3.67\u0026thinsp;\u0026plusmn;\u0026thinsp;0.49\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e216.88\u0026thinsp;\u0026plusmn;\u0026thinsp;13.03\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e336.31\u0026thinsp;\u0026plusmn;\u0026thinsp;46.84\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c6\"\u003e \u003cp\u003e84.86\u0026thinsp;\u0026plusmn;\u0026thinsp;16.99\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNC2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e3.67\u0026thinsp;\u0026plusmn;\u0026thinsp;0.43\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e192.42\u0026thinsp;\u0026plusmn;\u0026thinsp;28.84\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e331.74\u0026thinsp;\u0026plusmn;\u0026thinsp;34.04\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c6\"\u003e \u003cp\u003e92.90\u0026thinsp;\u0026plusmn;\u0026thinsp;14.07\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eM\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2.49\u0026thinsp;\u0026plusmn;\u0026thinsp;0.51*\u003csup\u003e#\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e169.85\u0026thinsp;\u0026plusmn;\u0026thinsp;11.55*\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e453.17\u0026thinsp;\u0026plusmn;\u0026thinsp;24.58*\u003csup\u003e#\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c6\"\u003e \u003cp\u003e84.58\u0026thinsp;\u0026plusmn;\u0026thinsp;11.59\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eFMT1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2.93\u0026thinsp;\u0026plusmn;\u0026thinsp;0.30*\u003csup\u003e#\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e179.44\u0026thinsp;\u0026plusmn;\u0026thinsp;36.56*\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e415.39\u0026thinsp;\u0026plusmn;\u0026thinsp;77.30*\u003csup\u003e#\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c6\"\u003e \u003cp\u003e88.37\u0026thinsp;\u0026plusmn;\u0026thinsp;3.33\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eFMT2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e3.88\u0026thinsp;\u0026plusmn;\u0026thinsp;0.11\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e205.31\u0026thinsp;\u0026plusmn;\u0026thinsp;17.07\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e325.93\u0026thinsp;\u0026plusmn;\u0026thinsp;41.37\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c6\"\u003e \u003cp\u003e82.94\u0026thinsp;\u0026plusmn;\u0026thinsp;6.60\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"6\"\u003eNote: Compared with NC1, *\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05; compared with NC2, \u003csup\u003e#\u003c/sup\u003e\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05; compared with M, \u003csup\u003ea\u003c/sup\u003e\u003cem\u003eP\u003c/em\u003e \u0026lt; 0.05; compared with FMT1, \u003csup\u003eb\u003c/sup\u003e\u003cem\u003eP\u003c/em\u003e \u0026lt; 0.05.\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec15\" class=\"Section2\"\u003e \u003ch2\u003e3.7 Serum TC and TG Levels\u003c/h2\u003e \u003cp\u003eLong-term high-fat diets caused lipid metabolism disorders. Serum TC and TG levels showed that: (1) TC levels: The M, FMT1, and FMT2 groups had significantly higher serum TC levels compared to the NC1 and NC2 groups (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05); the FMT1 and FMT2 groups had significantly lower TC levels compared to the M group (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05); no significant difference was observed between the FMT1 and FMT2 groups (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026gt;\u0026thinsp;0.05). (2) TG levels: The NC2, FMT1, and FMT2 groups had significantly lower serum TG levels compared to the NC1 group (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05); the M group showed a decrease without significant differences (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026gt;\u0026thinsp;0.05); the FMT1 group had significantly lower TG levels compared to the M group (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05); no significant differences were observed among the NC2, FMT1, and FMT2 groups (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026gt;\u0026thinsp;0.05).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab3\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eTC and TG Levels in Each Group of Rats (\u0026oline;\u003cem\u003ex\u003c/em\u003e\u0026thinsp;\u0026plusmn;\u0026thinsp;s,mmol/L)\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"4\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" 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 \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGroups\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003en\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eTC\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eTG\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNC1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1.34\u0026thinsp;\u0026plusmn;\u0026thinsp;0.22\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1.06\u0026thinsp;\u0026plusmn;\u0026thinsp;0.28\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNC2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1.11\u0026thinsp;\u0026plusmn;\u0026thinsp;0.17\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.70\u0026thinsp;\u0026plusmn;\u0026thinsp;0.18*\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eM\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2.26\u0026thinsp;\u0026plusmn;\u0026thinsp;0.39*\u003csup\u003e#\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.89\u0026thinsp;\u0026plusmn;\u0026thinsp;0.23\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eFMT1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1.69\u0026thinsp;\u0026plusmn;\u0026thinsp;0.14*\u003csup\u003e#a\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.61\u0026thinsp;\u0026plusmn;\u0026thinsp;0.15*\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eFMT2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1.67\u0026thinsp;\u0026plusmn;\u0026thinsp;0.17*\u003csup\u003e#a\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.65\u0026thinsp;\u0026plusmn;\u0026thinsp;0.12*\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"4\"\u003eNote: Compared with NC1, *\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05; compared with NC2, \u003csup\u003e#\u003c/sup\u003e\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05; compared with M, \u003csup\u003ea\u003c/sup\u003e\u003cem\u003eP\u003c/em\u003e \u0026lt; 0.05.\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e"},{"header":"4 Discussion","content":"\u003cdiv id=\"Sec17\" class=\"Section2\"\u003e \u003ch2\u003e\u003cb\u003e4.1\u003c/b\u003e The Impact of FMT on Gut Microbiota in Obese Rats\u003c/h2\u003e \u003cp\u003eGut microbiota is closely related to energy balance and metabolism of its host. Obesity is associated with changes in the relative abundance of the two main phyla in the gut microbiota, Bacteroidetes and Firmicutes \u003csup\u003e[\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e]\u003c/sup\u003e. The tendency toward obesity or increased body fat may be determined by the ratio of Bacteroidetes to Firmicutes, which can serve as a biomarker for diagnosing obesity-related metabolic syndrome \u003csup\u003e[\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e]\u003c/sup\u003e. Obesity is one of the indications for FMT in treating dysbiosis-related diseases \u003csup\u003e[\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e]\u003c/sup\u003e. FMT is considered as an effective means of reconstructing gut microbiota. Colonization of beneficial microbiota in the gut of obese rats can improve the gut environment and correct dysbiosis\u003csup\u003e[\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e]\u003c/sup\u003e. In this study, a high-fat diet increased the Firmicutes/Bacteroidetes ratio in rats. Normal fecal microbiota transplantation restored this ratio in obese rats to that of normal rats, while fecal microbiota transplantation from obese rats increased this ratio in normal rats. The abundance and diversity of gut microbiota in the M and FMT1 groups were lower than those in the NC1 group, suggesting that a high-fat diet alters the abundance and diversity of gut microbiota.\u003c/p\u003e \u003cp\u003eOverall, the structure of gut microbiota of the M and FMT1 groups was more similar, and was different from the NC1 group, indicating that transplantation of gut microbiota from obese rats could disrupt the gut microbiota of normal rats to some extent. Previous studies have found that Clostridium, a bacterium that can reduce fat content, increases in abundance to reduce fat content to some extent. If Clostridium abundance decreases, serum leptin levels increase. Supplementing probiotics to increase Clostridium abundance can enhance host sensitivity to leptin. The Ruminococcaceae family is also closely related to obesity, with its abundance significantly increased in the gut of high-fat diet-induced obese mice\u003csup\u003e[\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e]\u003c/sup\u003e, which was consistent with the results in this study. Akkermansia muciniphila, a Gram-negative bacterium, its colonization is generally believed to be negatively correlated with waist circumference, body weight, and body fat. It can reduce fat accumulation, enhance metabolism, and promote weight loss, with lower abundance in obese individuals\u003csup\u003e[\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e]\u003c/sup\u003e. However, in this study, Akkermansia muciniphila significantly increased in the high-fat groups, possibly due to the self-regulatory response of rats to a high-fat diet or the use of antibiotics. After neomycin-containing antibiotic intervention, Verrucomicrobia significantly increased in mice\u003csup\u003e[\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e]\u003c/sup\u003e. Oral vancomycin reduced nutrient absorption and increased Akkermansia muciniphila by regulating intestinal barrier function\u003csup\u003e[\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e]\u003c/sup\u003e. Streptomycin and vancomycin used in this study are narrow-spectrum antibiotics, and Akkermansia muciniphila is a dominant genus in the Verrucomicrobia phylum. Therefore, the increase in Akkermansia muciniphila may be related to antibiotic use.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec18\" class=\"Section2\"\u003e \u003ch2\u003e\u003cb\u003e4.2\u003c/b\u003e Normal Fecal Microbiota Exerts Effects through the SCFAs-GPR43-Gastrointestinal Peptide Pathway\u003c/h2\u003e \u003cp\u003eOne of the main ways to produce energy in the colon is through the metabolism of SCFAs, which increases lipid metabolic rates \u003csup\u003e[\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e]\u003c/sup\u003e. SCFAs not only serve as substrates for lipid and energy metabolism, but also act as regulatory factors to modulate host physiological metabolism \u003csup\u003e[\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e]\u003c/sup\u003e. An improper diet is the main cause of obesity and metabolic syndrome. When the diet changes, gut microbiota rapidly respond to these changes \u003csup\u003e[\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e]\u003c/sup\u003e. Diet can affect SCFA levels by altering the type and abundance of gut microbiota, thereby influencing obesity. SCFAs play a crucial role in maintaining gut health and function, providing energy and nutrients necessary for bacterial growth and reproduction. They maintain normal osmotic pressure, epithelial cell integrity, and promote epithelial cell repair in the gut \u003csup\u003e[\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e]\u003c/sup\u003e. SCFAs can exert various effects to maintain and improve gut health, control blood glucose homeostasis by regulating systemic energy balance, and have the potential to alleviate obesity and diabetes \u003csup\u003e[\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e]\u003c/sup\u003e. The results indicated that SCFA levels were significantly reduced in the M group, suggesting that microbiota transplantation exerts effects by influencing SCFA production. In addition to energy metabolism, SCFAs can act as signaling molecules, binding to G-protein-coupled receptors (GPCRs), stimulating GLP-1 secretion and PYY production, inhibiting gut motility, controlling appetite, altering transit time, and affecting lipid accumulation \u003csup\u003e[\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e]\u003c/sup\u003e. GPR43 is closely related to gut function and lipid metabolism, controlling the body\u0026rsquo;s energy utilization rate and maintaining a stable metabolic environment. Mice lacking GPR43 become obese under normal dietary conditions, while overexpression of GPR43 in mice prevents obesity even with a high-fat diet \u003csup\u003e[\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e]\u003c/sup\u003e. Studies indicate that a high-fat diet affects metabolism by reducing GPR43 expression. GPR43 can co-express with GLP-1, mediating SCFA-induced GLP-1 secretion. The fat-reducing effect of GLP-1 is widely studied and recognized, and GLP-1 levels are reduced in patients with obesity and metabolic syndrome \u003csup\u003e[\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e]\u003c/sup\u003e. As an incretin, GLP-1 plays a key role in regulating blood glucose levels and insulin secretion. GLP-1 agonists have been shown to reduce appetite, decrease energy intake, delay gastric emptying, and subsequently reduce body weight \u003csup\u003e[\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e]\u003c/sup\u003e, and also impact cardiometabolic risk factors, reducing the risk of cardiovascular events associated with obesity \u003csup\u003e[\u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e]\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eIncreased PYY production can reduce gut motility, increasing energy acquisition post-meal and enhancing satiety \u003csup\u003e[\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e]\u003c/sup\u003e. The results indicate that GLP-1 levels and PYY expression were lower in the M and FMT1 groups compared to the NC1 group. Reduced MTL can slow digestion and absorption, reducing postprandial hyperglycemia and hyperlipidemia \u003csup\u003e[\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e]\u003c/sup\u003e. Reduced motilin promotes gastric emptying, playing an important role in regulating gastrointestinal motility \u003csup\u003e[\u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e38\u003c/span\u003e]\u003c/sup\u003e. Although there were no significant differences in GAS levels among the groups, the secretion volume may not change significantly, and the response may be enhanced. The MTL levels in the M and FMT1 groups were higher than in the NC1 group, suggesting MTL activation in obese rats, increasing hunger by acting on the feeding center, and subsequently increasing food intake. Chen Luman et al. found through animal experiments that activation of the MTL receptor increases appetite and body weight of rats \u003csup\u003e[\u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e39\u003c/span\u003e]\u003c/sup\u003e. Reports indicate that MTL levels are higher in obese patients than in healthy individuals \u003csup\u003e[\u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e40\u003c/span\u003e]\u003c/sup\u003e. Stabilizing gastrointestinal hormone levels helps improve gastrointestinal function, consistent with the MTL expression results of this study.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec19\" class=\"Section2\"\u003e \u003ch2\u003e\u003cb\u003e4.3\u003c/b\u003e Pathology Conditions and Changes in Serum TC and TG\u003c/h2\u003e \u003cp\u003eIn M group, intestinal mucosal epithelial cells were disordered, with visible inflammatory cell infiltration, suggesting that a high-fat diet affects normal gastrointestinal digestion and excretion functions, leading to abnormal and pathological changes. Hepatocytes in M group were loosely and disorderedly arranged, with diffuse accumulation of lipid droplets, indicating that obese rats exhibited fatty liver and increased intra-abdominal fat. Due to the gut-liver axis, the liver is directly exposed to gut microbiota and their products. When gut microbiota is imbalanced, the permeability and integrity of the intestinal wall are affected, allowing a large number of bacterial products to enter the liver, leading to liver dysfunction \u003csup\u003e[\u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e41\u003c/span\u003e]\u003c/sup\u003e. An intact intestinal mucosal barrier and liver defense function are crucial for maintaining internal homeostasis. Numerous studies have shown that once the balance between the liver and the intestine is disrupted, it will lead to pathological changes such as hepatic steatosis and hepatitis \u003csup\u003e[\u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e42\u003c/span\u003e]\u003c/sup\u003e. A retrospective cohort study indicated that obesity is a significant risk factor for fatty liver disease, as the liver is a key site for lipid metabolism \u003csup\u003e[\u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e43\u003c/span\u003e]\u003c/sup\u003e. SCFAs can maintain the integrity of the intestinal barrier, preventing intestinal toxins like lipopolysaccharides from being absorbed through the intestinal epithelium and invading the liver. In colon epithelial cells, SCFAs bind to GPR43, activate caspase-1, and release interleukin-18, promoting intestinal epithelial repair \u003csup\u003e[\u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e44\u003c/span\u003e]\u003c/sup\u003e. Obesity damages the intestinal barrier, accelerating obesity, while SCFAs help improve the intestinal barrier \u003csup\u003e[\u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e45\u003c/span\u003e]\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eObese individuals have high visceral fat content, leading to the release of large amounts of free fatty acids into the liver, promoting TG synthesis. Excessive fat deposition can alter lipase activity, subsequently increasing TC levels \u003csup\u003e[\u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e46\u003c/span\u003e]\u003c/sup\u003e. Obese patients often exhibit altered blood lipid levels, typically showing elevated TG and TC levels. A high-fat diet increases fat deposition in rats, potentially accompanied by an overall increase in metabolic levels. On one hand, the intake of a high-fat diet requires the production of large amounts of bile acids for digestion and absorption, promoting the liver to accelerate TC breakdown to form bile acids. On the other hand, a high-fat diet can increase liver enzyme activity, speeding up TC synthesis. Based on the results of this study, it is speculated that the total metabolic level of TC in high-fat diet rats tends to increase, with the efficiency of TC synthesis exceeding that of breakdown, resulting in significant changes in serum TC levels in the FMT1 group and M group compared to the NC1 group. The TG levels in the FMT1 group and M group were lower than those in the NC1 group, possibly due to fasting for 12 hours before sampling, causing abnormal elevation of indicators in the NC1 group due to hunger, thus interfering with serum TG levels and causing inconsistent results compared to previous findings. The results of this study were consistent with revious reports\u003csup\u003e[\u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e47\u003c/span\u003e]\u003c/sup\u003e. Additionally, the TG levels in the antibiotic intervened NC2 group were lower than those in the NC1 group, suggesting that antibiotic intervention may also be related to the atypical TG levels observed in this study.\u003c/p\u003e \u003c/div\u003e"},{"header":"5 Conclusions","content":"\u003cp\u003eIn summary, a high-fat diet could induce related pathological reactions by affecting the gut microbiota-SCFAs-GPR43-gastrointestinal peptide pathway, lead to gut microbiota dysbiosis, SCFA metabolism disorders, and subsequent activation of GPR43 to release PYY, GLP-1, GAS, and MTL, result in lipid and energy metabolism disorders in the body. Normal microbiota can be colonized in the gut of obese rats through fecal microbiota transplantation, improving reduced abundance, diversity, and disordered microbiota structure, activating GPR43, initiating downstream mechanisms, regulating the secretion of peptide hormones by endocrine cells, and alleviating metabolic disorders induced by a high-fat diet. This suggests that gut microbiota can be a new target for treating obesity and other metabolic diseases, promoting a positive cycle of body metabolism. The SCFAs in this study included a combination of acetate, propionate, and butyrate, but their individual mechanisms in regulating obesity remain unclear. The specific pathways of different SCFAs\u0026rsquo; mechanisms of action, the synergy or contradictions of the same SCFA in different signaling pathways, and the synergy or contradictions of different SCFAs in the same signaling pathway could be explored in future studies.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis study was supported by the the Fundamental Research Funds for the Central Universities for the Unveiling of the List of Commander-in-Chief Programs in Beijing University of Chinese Medicine（2024-JYB-JBZD-038）;\u003c/p\u003e\n\u003cp\u003eNational Natural Science Foundation of China (81704121);\u003c/p\u003e\n\u003cp\u003eResearch on the Four Famous Doctors in Beijing and their Main Inheritors, and the Inheritance of the Experiences of Famous Doctors of Yanjing（BUCM-2023-JS-FW-077）;\u003c/p\u003e\n\u003cp\u003eHigh-level traditional Chinese medicine key subjects construction project of National Administration of Traditional Chinese Medicine\u0026mdash;Beijing University of Chinese Medicine,Chinese Medicine Epidemic Disease（No.zyyzdxk-2023264）\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthor Contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eHe Yu, Xiaomei Wang, and Xu Wang designed this study; Lijun Cui, Shaoyang Liu, Chen Bai, Jianhua Zhen,and Tiegang Liu have undergone the experiment; Xiaohong Gu, Xiaomei Wang, and Xu Wang provided experimental guidance; Lijun Cui, Xiaomei Wang, Xu Wang, and He Yu wrote a manuscript; HeYu and Xiaohong Gu funded this study.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConflict of Interest Statement\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eChen L, Cong D, Wang G, et al. 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Laboratory animal research,2015,31(4):166\u0026ndash;173.\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Obesity, High fat diet, Fecal microbiota transplantation, Intestinal flora-SCFAs-GPR43-gastrointestinal peptide","lastPublishedDoi":"10.21203/rs.3.rs-6307329/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-6307329/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cb\u003eObjective\u003c/b\u003e\u003c/p\u003e \u003cp\u003eTo explore the effect of high-fat diet and fecal microbiota transplantation (FMT) on \u0026ldquo;intestinal flora-SCFAs-GPR43-gastrointestinal peptide\u0026rdquo; pathway, and provide evidence and clues for the prevention and treatment of obesity caused by eutrophic diet.\u003c/p\u003e\u003cp\u003e\u003cb\u003eMethods\u003c/b\u003e\u003c/p\u003e \u003cp\u003e160 male SD rats were used in this study, 50 of them were randomly selected to be fed a standard rat diet, while the remaining 110 rats were fed a high-fat diet (D12492). After excluding the rats that did not meet the obesity criteria, the remaining rats were subjected to treatment with normal microbiota enema and obesity-associated microbiota enema. The rats were divided into normal control group 1 (NC1), normal control group 2 (NC2), obesity model group (M), obesity fecal microbiota transplantation group (FMT1), and normal fecal microbiota transplantation group (FMT2). The study observed the general situation, the index of liver, spleen and thymus in rats. Morphological changes of colon and liver tissues were examined under an optical microscope, and the alterations in gut microbiota were detected by 16s rDNA. Gas chromatography-mass spectrometry (GC-MS) was ued to measure the levels of short-chain fatty acids (SCFAs), including acetic acid, propionic acid, and butyric acid. Immunohistochemistry (IHC) was used to evaluate the expression of GPR43 in liver tissue. Additionally, gastrointestinal peptides in rat serum were quantified using the ELISA method, while cholesterol and triglyceride levels in serum were measured using an automatic biochemical analyzer.\u003c/p\u003e\u003cp\u003e\u003cb\u003eResults\u003c/b\u003e\u003c/p\u003e \u003cp\u003eThe high-fat diet successfully induced obesity rat models. This led to significant changes in gut environment and the survival environment of microbiota, such as Lactobacillus, reflecting the intestinal microecological disorders in rats with high-fat diet induced obesity, Different dietary interventions can lead to varing developments in gut microbiota. After antibiotic intervention, gut microbiota in rats were significantly suppressed, with reduced species diveristy and abundance, establishing an antibiotic-induced rat model. High-fat diet interventions resulted in significant changes in the relative abundances of specific gut bacterial species. Further analysis of microbial metabolites displayed that a high-calorie diet reduced the content of short-chain fatty acids (SCFAs) in feces, and subsequently reduced the expression of GPR43, resulting in improved abnormal expression of downstream gastrointestinal peptide.\u003c/p\u003e\u003cp\u003e\u003cb\u003eConclusions\u003c/b\u003e\u003c/p\u003e \u003cp\u003eHigh-fat diet affects the intestinal flora-SCFAs-GPR43-gastrointestinal peptide pathway, leading to related pathological reactions, such as intestinal flora imbalance and short-chain fatty acid metabolism disorders, which in return activates GPR43, and releases PYY, GLP-1, GAS, MTL, causing lipid and energy metabolism disorders in the body. Fecal microbiota transplantation (FMT) can colonize the intestinal tract of obese rats, improving the abundance, diversity and the structure of the flora, activating GPR43 and the downstream mechanisms. This regulation of peptide hormone secretion by endocrine cells can improve metabolic disorders caused by a high-fat diet and may play a significant role in preventing and treating obesity.\u003c/p\u003e","manuscriptTitle":"Effect of fecal microbiota transplantation on “intestinal flora-SCFAs-GPR43 - gastrointestinal peptide” pathway in rats with high-fat diet","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-04-24 07:41:07","doi":"10.21203/rs.3.rs-6307329/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"cc1e50ad-3ac6-4e21-b6e9-22f288cb82cd","owner":[],"postedDate":"April 24th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":46603047,"name":"Biological sciences/Microbiology/Bacteria"},{"id":46603048,"name":"Health sciences/Endocrinology/Endocrine system and metabolic diseases/Obesity"}],"tags":[],"updatedAt":"2025-09-03T14:27:13+00:00","versionOfRecord":[],"versionCreatedAt":"2025-04-24 07:41:07","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-6307329","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-6307329","identity":"rs-6307329","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}