The Intestinal Microbiota Modulates the Visceral Sensitivity Involved in IBS Induced by Restraint Combined with Tail Clustering

Na Deng ¹ Shiqin Xie ¹ Qin Liu ¹ Huiyi Peng ¹ Leyao Fang ¹ Junxi Shen ¹ Xiaoyuan Lin ^2*

• ¹ School of Chinese Medicine, Hunan University of Chinese Medicine,, hunan changsha, China
• ² The First Hospital of Hunan University of Chinese Medicine, Changsha, Hunan, China, School of Chinese Medicine, Hunan University of Chinese Medicine,, hunan changsha, China

Objective: To compare three common stimuli that induce emotional stress to identify the optimal method for establishing an animal model that aligns with the clinical pathogenesis of irritable bowel syndrome (IBS) and to explore the gut microbiota mechanisms underlying IBS development.Methods: The NC group was fed normally . The BM, CTM, and BCTM groups were administered restraint, tail clamping, and restraint combined with tail clamping stimulation, respectively, and the modeling duration was 7 days. General behavioral changes in the mice were observed. The pain threshold, gastric residue rate, small intestine propulsion rate, and serum levels of SCFAs、5-HT、IL-10、TNF-α were assessed. Histopathological analysis was performed. 16S rRNA high-throughput sequencing was subsequently conducted. Results:Compared with the NC group, the BM, CTM, and BCTM groups of mice presented varying degrees of emotional hyperreactivity, accompanied by significantly reduced food intake and fecal water content and markedly elevated levels of inflammation, all of which are indicative of IBS symptoms. Among them, the BCTM group presented the most pronounced emotional hyperreactivity and irritability. The mice in the BCTM group had significantly higher gastric residue rates and 5-HT levels, with a marked reduction in pain tolerance. The gut microbiota of the mice in the BM, CTM, and BCTM groups all exhibited dysbiosis, with changes in the diversity, structural composition, and function of the microbial community. Specific bacterial taxa were enriched in each stress group, and their corresponding KEGG pathways were also significantly altered. Correlation analysis revealed that SCFAs were significantly positively correlated with the small intestine propulsion rate, whereas 5-HT was positively correlated with the gastric residue rate and negatively correlated with the pain threshold. SCFAs were positively correlated with IL-10 and TNF-α, and 5-HT was significantly positively correlated with IL-10 and TNF-α. In the BCTM group, the characteristic bacteria Acinetobacter and Akkermansia were significantly correlated with SCFAs and 5-HT.Conclusion: The restraint combined with the tail clamp stress method is superior among the three stress protocols and successfully induces the IBS mouse model. Acinetobacter and Akkermansia may contribute to the development of IBS induced by restraint combined with tail clamp stress through the regulation of SCFAs and 5-HT.