16S rRNA高通量测序研究柴芪汤对代谢综合征大鼠肠道菌群的影响

彭龙 张立平

摘要 目的：采用16S rRNA高通量测序技术分析柴芪汤对代谢综合征（MS）大鼠肠道菌群的影响。方法：通过Illumina Miseq PE300型高通量测序仪对大鼠粪便肠道菌群进行16S rRNA基因V3-V4可变區检测，运用Usearch、Mothur、LEfSe分析肠道菌群的组成结构及不同菌属相对丰度变化。结果：柴芪汤能够逆转MS大鼠肠道菌群物种多样性的下降，并纠正MS导致的菌群组成结构改变。研究发现MS大鼠的肠道菌群在菌属水平有57个菌属相对丰度发生显著变化，而柴芪汤能将其中的54个菌属恢复至正常水平。结论：从肠道微生态角度阐释了MS的可能病机，柴芪汤可以通过调节肠道菌群防治MS。

关键词 代谢综合征;柴芪汤;健脾疏肝;肠道菌群;16S rRNA高通量测序;炎症反应;短链脂肪酸;脂多糖

Effects of Chaiqi Decoction on Gut Microbiota in MS Rats Based on 16S rRNA

High-Throughput Gene Sequencing Study

PENG Long，ZHANG Liping

（Beijing University of Chinese Medicine，Beijing 100029，China）

Abstract Objective：To explore the effects of Chaiqi Decoction on gut microbiota in metabolic syndrome rats based on high-throughput 16S rRNA gene sequencing.Methods：Illumina Miseq PE300 high-throughput sequencer was used to detect the 16S rRNA gene V3-V4 variable region of rat fecal intestinal flora.Usearch，Mothur，and LEfSe were used to analyze the composition and structure of intestinal flora and the relative abundance of different bacteria.Results：Chaiqi Decoction can reverse the decline in the species diversity of the intestinal flora of rats with metabolic syndrome and correct the changes in the composition and structure of the flora caused by the metabolic syndrome.The study found that the relative abundance of 57 genera of the intestinal flora of rats with metabolic syndrome changed significantly at the level of genus，and Chaiqi Decoction could restore 54 of them to normal levels.Conclusion：The possible pathogenesis of metabolic syndrome from the perspective of intestinal microecology is explained.Chaiqi Decoction could prevent and treat metabolic syndrome by regulating the intestinal flora.

Keywords Metabolic syndrome; Chaiqi Decoction; Invigorating spleen and soothing liver; Gut microbiota; 16S rRNA high-throughput gene sequencing; Inflammation reaction; Short-chain fatty acids; Lipopolysaccharide

中图分类号：R285.5文献标识码：Adoi：10.3969/j.issn.1673-7202.2021.05.014

代谢综合征（Metabolic Syndrome，MS）是一组复杂的代谢紊乱症候群，包括糖代谢异常、腹型肥胖、血脂代谢异常和高血压，是心脑血管疾病、慢性肾病、非酒精性脂肪肝等疾病的重要危险因素之一[1]。近年来研究发现，肠道菌群是影响人体物质能量代谢的重要因素，同时也是导致炎症介质过度表达、胰岛素抵抗，并进一步引起MS重要原因[2]，而中医学认为脾胃主运化，与肠道菌群功能相符，因此从调理脾胃即调节肠道菌群入手改善MS患者的代谢状态成为近年来的研究热点。张立平教授认为，肝失疏泄、脾失健运状态导致物质代谢能力下降是其发病的核心病机[3]，并提出应用健脾疏肝调理脾胃法治疗MS，拟方“柴芪汤”。前期研究中课题组已经证实柴芪汤能够纠正代谢异常状态，改善MS大鼠的胰岛素抵抗，减轻血管内皮损伤[4-10]。……