16s rRNA测序联合代谢组学技术探究逍遥抑激汤治疗IBS-D的作用机制

To explore the mechanism of Xiaoyao Yiji decoction in the treatment of IBS-D by the 16s rRNA sequencing combined with metabolomics

摘要:

目的 通过 16s rRNA 测序联合代谢组学探讨逍遥抑激汤治疗腹泻型肠易激综合征（diarrhea-predominant irritable bowel syndrome，IBS-D）的作用机制。方法 招募10例就诊于陕西省中医医院且符合纳入标准的IBS-D患者，收集患者逍遥抑激汤治疗前后的粪便样本，进行16s rRNA测序，多样性分析肠道菌群组成和群落结构的差异，筛选差异性菌群；通过粪便代谢组学分析筛选分析差异性表达代谢产物和通路。结果 IBS-D患者在逍遥抑激汤治疗后，门水平上肠道菌群中的厚壁菌门、变形菌门、放线菌门、拟杆菌门发生明显改变，厚壁菌门明显上调，属水平上巨单胞菌、志贺氏埃希氏菌、双歧杆菌、普拉梭菌是优势菌群，多样性分析表明治疗后肠道菌群物种丰富度减低，但均匀度和多样性均增加，群落结构和组成基本一致，Negativicutes、韦荣球菌属是组间具有统计学差异的特异性菌群。筛选出21种疾病相关的差异代谢产物，涉及16条代谢相关通路，苯丙氨酸代谢影响最显著。结论 逍遥抑激汤可能通过调控厚壁菌门、韦荣球菌等肠道菌群，调控苯丙氨酸代谢和组氨酸代谢等通路以达到治疗IBS-D的作用。

Abstract:

Objective To investigate the mechanism of action of Xiaoyao Yiji decoction in the treatment of Diarrhea-predominant irritable bowel syndrome（IBS-D） by 16s rRNA sequencing combined with metabolomics.Methods A total of 10 IBS-D patients who met the inclusion criteria in the outpatient clinic of the Department of Shaanxi Hospital of Chinese Medicine were enrolled，feces samples were collected before and after treatment by Xiaoyao Yiji decoction，the differences of intestinal flora composition and community structure，and screened differential flora were analyzed by the 16S rRNA. The differential expressed metabolites and pathways were screened and analyzed by fecal metabolomics analysis.Results Xiaoyao Yiji decoction can significantly regulate the abundance of intestinal firmicutes，proteobacteria，actinomycetes，Bacteroides，Megamonas，Escherichia shigella，Bifidobacterium，P. pristrilla，and other flora in IBS-D patients and identify 21 disease-related differential metabolites involving 16 metabolism-related pathways，among which the pathway affects phenylalanine metabolism the most and histidine metabolism followed. With the Xiaoyao Yiji decoction treatment，the gut microbiota in IBS-D patients had been changed as Firmicutes，Proteobacteria，Actinobacteria，Bacteroidetes in the phylum level and Firmicutes increased significantly. Monelensis，Escherichia-Shigella，Bifidobacterium，Faecalibacterium，Blautia were the dominant flora. The diversity analysis showed that the species richness of the intestinal flora group decreased after the treatment，but the evenness and diversity increased，and the community structure and composition were basically consistentdiversity. Negativicutes，Veillonellales_Selenomonadales is specific flora with statistical differences between groups. Twenty-one disease-related differential metabolites were selected，involving in 16 metabolism-related pathways，with the most prominent effects on phenylalanine metabolism.Conclusion Xiaoyao Yiji decoction may achieve the therapeutic effect of IBS-D by regulating the intestinal flora of Firmicutes and Veillonellales，regulating phenylalanine metabolism and histidine metabolism pathways.