基于网络药理学和16S rRNA测序探讨黄连解毒汤治疗2型糖尿病的作用机制OA

Objective:To investigate the mechanism of action of Huanglian Jiedu Decoction in the treatment of type 2 diabetes mellitus by network pharmacology and 16S rRNA sequencing methods.Methods:The active ingredients and targets of Huanglian Jiedu Decoction were screened in the TCM Systematic Pharmacology Database and Analysis Platform(TCMSP),and the ob-tained targets were imported into the protein database(UniProt)to find the corresponding gene names.Disease targets were col-lected in the human genome database(GeneCards),the online human Mendelian genetic database(OMIM),and the TTD data-base.The intersection targets of drugs and diseases were obtained through the Venn diagram,and the"drug ingredient-disease tar-get"network was constructed by using Cytoscape 3.7.2 software to determine the key active ingredients.Protein-protein interaction(PPI)analysis was performed through the STRING database,and key targets were screened using the plug-in function in Cyto-scape 3.7.2 software.Then,the KEGG enrichment analysis was carried out through the Metascape database,and the"drug component-disease target-signaling pathway"network was constructed to identify the key signaling pathways.Finally,the network pharmacology results were verified by H&E staining and 16S rRNA sequencing.Results:A total of 85 active ingredients and 236 corresponding targets were found in Huanglian Jiedu Decoction,and 1 549 disease targets were found by network pharmacology analysis.There were 133 intersecting targets and 62 corresponding active ingredients.The key active ingredients in Huanglian Jie-du Decoction are quercetin and wogonin,etc.The core targets of the PPI network analysis were protein kinase 1(AKT1),tumor necrosis factor(TNF),interleukin-6(IL-6),and phosphoinositide 3-kinase/protein kinase B(PI3K/Akt)signaling pathways are closely related.Through animal experiments,it was found that compared to the normal group,the colon tissue structure of the rats in the model group was abnormal,the glands were atrophied and disordered,the spacing between some glands was widened,and the goblet cells were significantly reduced.After the intervention of Huanglian Jiedu Decoction,the tissue structure of the co-lon was significantly improved,the glands were arranged neatly,and a large number of goblet cells could be seen.In addition,Huanglian Jiedu Decoction could affect the richness and diversity of intestinal microbiota,increasing the abundance of Firmicutes at the phylum level,decreasing the abundance of Bacteroides,and increasing the abundance of beneficial bacteria such as Lactoba-cillus,Allobaculum and Blautia at the genus level.LEfSe analysis showed that Allobaculum,Lactococcus and Blautia were dif-ferential markers in the treatment of type 2 diabetes mellitus by Huanglian Jiedu Decoction.Conclusion:Huanglian Jiedu Decoc-tion can promote the proliferation of beneficial bacteria,reduce intestinal inflammation,and repair the intestinal barrier to treat type 2 diabetes.