Objective:To explore the effects of Jianpi Mixture on the glucose receptor/glucagon like peptide-1 (GLP-1)/5-hydroxytryptamine (5-HT) signaling pathway and the mechanism of Jianpi Mixture in improving visceral hypersensitivity in diarrhea-predominant irritable bowel syndrome (IBS-D) rats.Methods:Totally 40 healthy male SPF rats were divided into blank group, model group, Jianpi Mixture low- and high-dosage groups according to the random number table method, with 10 rats in each group. Except for the blank group, the rest of the rats were prepared by the "senna gavage combined with restraint stress" method to replicate the IBS-D spleen deficiency model. The aqueous extract of Jianpi Mixture was 2.52 and 7.56 g/kg in the low- and high-dosage groups, and the same volume of normal saline was given to the blank group and the model group, once a day, for 14 days. On the 1st, 7th and 14th days of administration, the body weight of rats was recorded, the fecal traits of the rats were scored with Bristol score, the visceral sensitivity of IBS-D rats was evaluated by the abdominal wall retraction reflex score (AWR) under rectal dilation, the morphology of colonic mucosal tissues was observed by HE staining, and the protein expressions of T1R2, SLGT1, GLUT2, GLP-1 and 5-HT in colon tissues were detected by immunohistochemical staining.Results:On the 1st, 7th and 14th days of administration, compared with the model group, the weight of the rats in the Jianpi Mixture high-dosage group increased ( P<0.05); on the 14th day of administration, the fecal Bristol score decreased significantly ( P<0.05) and the AWR score increased ( P<0.05); HE staining showed that the mucosal epithelium of colon tissue in the model group was slightly damaged, and local edema was visible. The pathological condition of colonic mucosa in the Jianpi Mixture low- and high-dosage groups was effectively improved; compared with the model group, the expressions of T1R2 and GLP-1 in the Jianpi Mixture low- and high-dosage groups increased ( P<0.05), and the expression of 5-HT decreased ( P<0.05); the protein expressions of GLUT2 and SGLT1 in the Jianpi Mixture high-dosage group increased ( P<0.05). Conclusion:Jianpi Mixture can treat spleen-deficiency IBS-D by restoring the normal expression of intestinal glucose receptors, regulating the secretion of GLP-1/5-HT, improving intestinal motility and alleviating visceral hypersensitivity.

The intestinal flora and gut barrier function are of great significance for gut function and human health. When the intestinal flora is disrupted and the gut barrier structure is disrupted, it can lead to bacterial translocation, endotoxin influx into the bloodstream, and the production of pro-inflammatory factors, leading to various tissue damage in the body. Tongfu method in TCM can affect the intestinal environment by regulating intestinal permeability and immune response, restoring normal intestinal movement, and regulating the structure and metabolites of intestinal flora, thereby maintaining intestinal homeostasis and body health. The research on regulating intestinal flora and improving intestinal barrier function by Tongfu method can provide reference for further research on the relationship between TCM and intestinal microecology, and provide ideas for clinical treatment.

Objective To observe the difference in the expression of micro-ribonucleic acid(miRNA)in the colon tissue of irritable bowel syndrome-diarrhea(IBS-D)rats and rats treated with Jianpi Mixture,and predict The miRNA and corresponding genes involved in the treatment of IBS-D with Jianpi Mixture,providing a basis for finding potential therapeutic targets for IBS-D.Methods The rats were randomly divided into a blank group,a model group and a traditional Chinese medicine group,with 8 rats in each group.Both the model group and the Chinese medicine group used acute and chronic stimulation + senna leaf gavage to make the IBS-D model.The Chinese medicine group was gavaged with Jianpi Mixture after model building for 14 days.General condition,body weight and Bristol score were observed and recorded.Hematoxylin-eosin(HE)staining was used to observe the histopathological changes of colon in each group of rats.Transcriptome sequencing of rat colon tissue in each group to screen and map differentially expressed miRNAs.Perform GO(Gene oncology)functional enrichment analysis and KEGG(Kyoto Encyclopedia of Genes and Genomes)pathway enrichment analysis on the target genes corresponding to the screened common miRNAs.Results Compared with the model group,the weight of the rats in the Chinese medicine group increased significantly(P<0.01),and the Bristol score of the feces decreased significantly(P<0.01).The pathological damage of the colon tissue of rats in the Chinese medicine group was significantly reduced.Compared with the blank group,there were 109 differentially expressed genes in the model group,of which 80 were up-regulated and 29 were down-regulated.Compared with the model group,there were 109 differentially expressed genes in the traditional Chinese medicine group,of which 19 were up-regulated and 90 were down-regulated.Mapping the genes of the two comparisons found that 74 miRNAs in the model group were increased and 7 were decreased.The changes of these miRNAs were all reversed after the intervention of traditional Chinese medicine.The GO function enrichment analysis of target genes showed that Jianpi Mixture was mainly involved in synaptic vesicle positioning and transport,cytoplasmic transport,negative regulation of growth and development,and sugar transport after acting on rats.KEGG pathway enrichment analysis showed that MAPK,Wnt,Hippo,TNF,oxygen toxin,cAMP and other pathways were enriched.Conclusion After the intervention of Jianpi Mixture,IBS-D-related miRNA expression profiles have changed significantly;Jianpi Mixture may play a protective role in the treatment of IBS-D by regulating the MAPK signaling pathway and Wnt signaling pathway.Screening the core genes and predicting the miRNAs interacting with them provide a theoretical basis for the study of the pathogenesis of IBS-D and the therapeutic targets of Jianpi Mixture.