基于16S rRNA技术与代谢组学探究茵陈蒿汤治疗非酒精性脂肪肝的作用机制

Mechanism of Yinchenhao Decoction in treating non-alcoholic fatty liver based on 16S rRNA technique and metabonomics

目的:联合16S rRNA技术与肝脏代谢组学探究茵陈蒿汤治疗非酒精性脂肪肝(NAFLD)的作用机制,并找寻相关肠道菌群、代谢产物及通路的关联性。方法:采用高脂饮食诱导大鼠建立NAFLD模型,造模成功后,给予茵陈蒿汤进行治疗2周。利用Illumina Miseq平台对大鼠16S rRNA V4区的肠道菌群进行测序,结合肝脏代谢组学及Pearson系数法构建差异菌群、代谢产物及通路的互作体系。结果:茵陈蒿汤可改善NAFLD模型大鼠厚壁菌门、拟杆菌门、放线菌门及变形菌门多样性的失调,并对其中16种菌属具有显著性的回调作用。鉴定出13个与治疗相关的肝脏代谢产物,涉及到最强的3个代谢通路分别为甘油磷脂代谢、嘌呤代谢和谷胱甘肽代谢。同时,发现具有强相关作用的肠道菌属与潜在生物标志物的互作关联性。结论:茵陈蒿汤通过干预相关肠道菌群与肝脏组织的代谢物及通路对NAFLD起到治疗作用,16S rRNA技术与代谢组学的联合分析可应用于对此类疾病的病理机制进行评价。

Objective:To explore the mechanism of action of Yinchenhao Decoction in the treatment of non-alcoholic fatty liver,and to find the correlation of relevant intestinal flora,metabolites and pathways by 16 s rRNA technology combined with metabonomics.Methods:High-fat diet was used to induce the NAFLD model of rats.After the model was established successfully,the rats were treated with Yinchenhao Decoction.The intestinal flora of rats in the 16 s rRNA V4 region was sequenced by Illumina Miseq platform,and the interaction system of different flora,metabolites and pathways was constructed by combining liver metabonomics and Pearson coefficient method.Results:Yinchenhao Decoction could improve the diversity disorder of firmicutes,bacteroides,actinobacteria and proteobacteria in NAFLD model rats,and has a significant callback effect on 16 species of bacteria.Thirteen liver metabolites related to treatment were identified,and the three strongest metabolic pathways were glycerophosphatide metabolism,purine metabolism and glutathione metabolism.At the same time,the strong correlation between intestinal bacteria and potential biomarkers was found.Conclusion:Yinchenhao Decoction has a therapeutic effect on NAFLD by intervening the metabolites and pathways of relevant intestinal flora and liver tissues.The combined analysis of 16 s rRNA technique and metabolomics can be used to evaluate the pathological mechanism of such diseases.