Effects of Jianpi Huashi Prescription and Its Different Components on Intestinal Microbiota in Rats Model of D-IBS

Xifeng LI¹Yu-Ling Liu²Ting-Ting Pan¹Nan Hu²Chun-Lei Du²Jianyu Zhou²Pei Liu²Ying-Han Wang^2*

• ¹Institute of Chinese Materia Medica, Chengde Medical University, Hebei Chengde, 067000, China, Chengde Medical University, Chengde, China
• ²Key Laboratory for Research and Development of Chinese Traditional Medicine of Hebei Province，Institute of Chinese Materia Medica, Chengde Medical University, Hebei Chengde, 067000, China, Chengde Medical University, Chengde, China

BACKGROUND: Diarrhea-predominant irritable bowel syndrome (D-IBS) is a clinically prevalent functional gastrointestinal disorder, categorised in Traditional Chinese Medicine (TCM) "diarrhea", "abdominal pain", and "depression syndrome" patterns. The gut microbiota plays a pivotal role in its pathogenesis. Jianpi Huashi prescription, a TCM formulation with demonstrated therapeutic efficacy, is widely employed in D-IBS treatment; however, its mechanism of action on gut microbiota remains incompletely understood. In this study, 16S rRNA sequencing was utilised to analyse gut microbial composition. Furthermore, the effects of different prescription components were investigated from a gut microbiota-centric perspective.To investigate the mechanism of Jianpi Huashi prescription and its constituent components in treating D-IBS from perspective of intestinal microbiota.Ninety male rats were randomly allocated into nine groups: normal control group (ZC), model group(M), positive control group (PC), Jianpi Huashi prescription groups (low [DJL], medium [ZJL], and high [GJL] doses), Qushi group (QS), Buyi (BY) group, and Liqirougan group (LQRG). All rats except the ZC group were received senna extract via oral administration and combined with restraint stress to simulate the pathological state of D-IBS with liver depression and spleen deficiency syndrome. Then, except for the normal group and model group, each group received the intervention treatment of corresponding drugs for 14 days.Visceral sensitivity was assessed using the abdominal withdrawal reflex (AWR) test.Subsequent analysis employed LC-MS targeted metabolomics to quantify short-chain fatty acid (SCFA) concentrations and 16S rRNA sequencing to characterise gut microbiota composition.Linear Discriminant Analysis Effect Size (LEfSe) was used to analyse the flora changes of rats in each group from door to genus, and the species with significant differences were also analysed. therapeutic efficacy, comprising ingredients that tonify the spleen and kidney, regulate qi, soften the liver, and eliminate dampness, is well-established employed in D-IBS management.Nonetheless, its mechanistic effects on gut microbiota remain incompletely elucidated. Our research group has demonstrated that Jianpi Huashi Granule can decrease the levels of 5-HT and its receptors in the colon mucosa of rats while increasing the levels of CGRP. This study aimed to elucidate the therapeutic mechanisms of the Jianpi Huashi prescription and its constituent components through 16S rRNA sequencing-based gut microbiota analysis.