不同中医证型2型糖尿病患者肠道菌群差异研究  

Study on Difference of Intestinal Flora in Patients with Type 2 Diabetes with Different TCM Syndromes

作　　者：殷美琦[1,2] 刘元君 牟宗平[1] 金丽娟 江国荣 黄菲[1,2] YIN Meiqi;LIU Yuanjun;MOU Zongping;JIN Ljuan;JIANG Guorong;HUANG Fei(Suzhou Traditional Chinese Medicine Hospital Affiliated to Nanjing University of Chinese Medicine,Suzhou 215009,Jiangsu,China;Clinical Pharmaceutical Laboratory of Traditional Chinese Medicine,Suzhou Academy of Wumen Chinese Medicine,Suzhou 215009,Jiangsu,China)

机构地区：[1]南京中医药大学附属苏州市中医医院,江苏苏州215009 [2]苏州市吴门医派研究院中药临床药学实验室,江苏苏州215009

出　　处：《辽宁中医杂志》2023年第4期1-7,I0001,共8页Liaoning Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金项目(81973740);苏州市科技发展计划(民生科技)项目(SS202006);苏州市姑苏卫生人才培养项目(GSWS2020079);苏州市“临床医学专家团队”引进项目(SZYJTD201718)。

摘　　要：目的探讨不同中医证型2型糖尿病(T2DM)患者肠道菌群差异。方法选择住院T2DM患者200例,按中医证型分为阴虚热盛证40例,湿热困脾证33例,气阴两虚证103例,阴阳两虚证24例,同时选取健康体检者26名作为健康对照,收集临床资料,采集血清标本测定糖脂代谢等指标,采集粪便标本采用16S rRNA高通量测序技术进行测序。采用R 3.4.1软件分析肠道菌群组成和Alpha、Beta多样性指标,并进行差异性分析,使用LEfSe方法挖掘不同证型标志肠菌并分析与临床指标相关性,PICRUSt预测肠道菌群功能变化特征。结果T2DM各证候组厚壁菌门与拟杆菌门的比例均低于健康对照组。与健康人群相比,T2DM患者肠道菌群多样性降低(P<0.05),湿热困脾证肠道菌群多样性最低。Venn图分析发现T2DM气阴两虚证有70个独有OTUs,阴虚热盛证、湿热困脾证和阴阳两虚证独有的OTUs分别是13、4和8个。MRPP分析显示T2DM不同证候肠道菌群结构组间差异大于组内差异(P<0.05)。属水平物种显著性差异分析显示,4种证候之间具有统计学意义的菌属共有16种。LEfSe分析显示在T2DM的4种证候中,阴阳两虚证多样性最突出,拥有8种代表菌,其次是气阴两虚证及阴虚热盛证,各有2种代表菌;代表菌与临床指标相关性分析显示与病程、体质量指数(BMI)、脂代谢指标等存在相关性;湿热困脾证贯穿糖尿病全程,与各证型存在重叠,无代表菌群。菌群基因功能预测也证实4种不同证候肠道菌群功能在糖类物质代谢和氨基酸代谢等途径存在统计学意义(P<0.05)。结论不同中医证型T2DM患者存在菌群结构差异,可能影响着糖尿病病程进展及不同中医证候的发生、发展及结局。Objective To explore the differences of intestinal flora in patients with type 2 diabetes mellitus(T2DM)with different TCM syndromes through the high-throughput 16S rRNA sequencing analysis.Methods According to TCM syndrome types,200 inpatients with T2DM were divided into Yin deficiency and heat excess syndrome(40 cases),damp-heat accumulating spleen syndrome(33 cases),QI-YIN deficiency syndrome(103 cases)and Yin-Yang deficiency syndrome(24 cases).At the same time,26 healthy patients were also involved as the healthy control group.The clinical data,serum and fecal specimens were collected,and the fecal specimens were sequenced by 16s rRNA high-throughput sequencing technique.The software R3.4.1 was used to analyze the composition and diversity of intestinal flora.LEfSe method was used to compare intestinal flora markers differences and analyze the correlation of these markers and clinical data.The change characteristics of intestinal flora function were predicted by the PICRUSt.Results The ratio of Firmicutes to Bacteroidetes in each T2DM syndrome group was lower than that in the healthy control group.The intestinal flora diversity of T2DM patients was significantly different from that of the healthy control group(P<0.05),and that of the damp-heat accumulating spleen syndrome group was the lowest.Venn diagram analysis showed that 70 OTUs only appeared in the Qi-Yin deficiency syndrome group,while 13,4 and 8 OTUs in Yin deficiency and heat excess syndrome group,damp-heat accumulating spleen syndrome group and Yin-Yang deficiency syndrome group,respectively.MRPP analysis showed that the difference of intestinal flora structure in different syndromes of T2DM among groups was greater than that within groups(P<0.05).The significant difference analysis of species at the level of genus showed that there were 16 genera with statistical differences among the four groups.LEfSe analysis showed that among the four groups of T2DM,Yin-Yang deficiency syndrome group showed much more diversity,with 8 representative intestinal fl

关 键 词：2型糖尿病 中医证候 肠道菌群 16S rRNA测序 

分 类 号：R698.2[医药卫生—泌尿科学]