16S rDNA基因测序分析阳虚质人群肠道菌群特征的研究

Study on Characteristics of Intestinal Flora in Yang Deficiency Constitution through rDNA Gene Sequencing Analysis

目的:研究阳虚质和平和质肠道微生物菌落构成变化,探讨阳虚质肠道菌群特点。方法:选取阳虚质及平和质人群,每组各30例,提取两组人群新鲜粪便样本中细菌DNA,运用Illumina MiSeq高通量测序平台对样本DNA进行338F806R区16S rDNA测序以检测肠道菌群。应用I-Sanger生信云平台分析门水平和属水平的肠道菌群结构变化。结果:PLS-DA显示阳虚质与平和质在肠道菌群组成上差异有统计学意义(P<0.05)。阳虚质Alpha多样性有下降趋势。在门水平上两组优势菌门都是厚壁菌门、变形菌门、拟杆菌门、放线菌门,阳虚质拟杆菌门/厚壁菌门的比值低于平和质。属水平上与平和质相比,阳虚质中粪栖杆菌、直肠真杆菌属、拟杆菌属、巨单胞菌属、罗氏菌属丰度下降。埃希氏杆菌/志贺氏杆菌属、链球菌属、消化链球属、柯林斯菌属等菌属丰度上升。阳虚质拟杆菌丰度低于平和质,差异具有统计学意义(P<0.05)。结论:阳虚质多样性呈下降趋势,丰度最高的菌门为厚壁菌门,其次为变形菌门,拟杆菌属相对丰度的显著下降是阳虚质肠道菌群的特征。

Objective:To study the changes to intestinal microbial composition of yang deficiency constitution,and to explore the characteristics of intestinal flora in yang deficiency constitution.Methods:People of yang deficiency constitution and of balanced constitution were recruited as 2 groups,30 people in each group.The bacterial DNA from fresh stool samples were extracted,and were subjected to 16S rDNA sequencing of the 338F_806R region using the Illumina MiSeq high-throughput sequencing platform to detect intestinal flora.The I-Sanger Bio-cloud platform was used to analyze the structural changes of the intestinal flora at the level of phylum and genus.Results:PLS-DA showed a significant difference in the composition of the intestinal flora between the yang deficiency constitution and the balanced constitution.The alpha diversity of yang deficiency showed a downward trend.At the level of phylum,the two most abundant abundances were Phylum Firmicutes,Proteobacteria,Bacteroidetes and Actinobacteria,and the ratio of Bacteroides/Phylum Firmicutes in the yang deficiency constitution was lower than that in the balanced constitution.At the genus level,yang deficiency constitution was lower than the balanced constitution on the abundance of faecalis,megamonas,rosenbergia,erectal bacilli,bacteroides,while higher on the abundance of the Escherichia/Shigella,Streptococcus,Peptostreptococcus and Collins.On the abundance of the Bacteroides,the yang deficiency type was lower than the balanced constitution(P<0.01).Conclusions:The diversity of yang deficiency constitution is decreased.Phylum with the highest abundance is the Phylum Firmicutes,followed by the Proteobacteria.The obvious decrease of the abundance of Bacteroides is the characteristic of intestinal flora in the yang-deficiency constitution.