基于16S rDNA技术分析化瘀温胆汤对糖耐量受损大鼠肠道菌群的影响

Effects of Huayu Wendan Decoction on Intestinal Flora of IGT Rats Based on 16S rDNA

目的基于16S rDNA技术分析糖耐量受损(IGT)大鼠肠道菌群结构特征,初步探讨化瘀温胆汤可能的干预机制。方法70只雄性SD大鼠中随机抽取10只作为空白组,其余60只予高脂高糖饲料喂养21周构建IGT大鼠模型,抽取30只IGT模型大鼠随机分为模型组、化瘀温胆汤组和二甲双胍组,每组10只。化瘀温胆汤组给予中药水煎剂灌胃,二甲双胍组给予二甲双胍悬浊液灌胃,空白组及模型组给予生理盐水灌胃。药物干预4周后测定大鼠空腹血糖(FPG)及灌胃葡萄糖溶液后2 h血糖(2 h PG),采集结肠内粪便,利用16S rDNA技术,基于OTU聚类结果,分析样本菌群的多样性和丰度,同时分析各样本在门、属水平的物种组成。结果基于OTU的韦恩图结果显示,有445个OTU为4组共有,4组间相似性较高,其中空白组和模型组相似性最低。α多样性分析显示,与空白组比较,模型组α多样性指数下降,化瘀温胆汤组、二甲双胍组α多样性指数显著升高。菌群均匀度和菌群丰度从高到低依次为空白组、化瘀温胆汤组、二甲双胍组、模型组。物种相对丰度结果显示,门水平各样本以厚壁菌门(Firmicutes)为优势菌,Saccharibacteria在空白组数量最多,模型组数量最少,广古菌门(Euryarchaeota)在空白组数量最多,模型组该门几乎为零;属水平各样本以乳酸菌属(Lactobacillus)数量最多,Lactobacillus在空白组数量最少,模型组数量最多。群落结构差异性分析显示,组间两两比较,肠道菌群分布具有明显差异性。结论IGT大鼠肠道菌群稳态失衡,菌群多样性及丰度在一定程度上降低,化瘀温胆汤可能通过提高肠道菌群多样性和相对丰度影响IGT大鼠肠道菌群稳态。

Objective To analyze the structural characteristics of intestinal flora in rats with impaired glucose tolerance(IGT)based on 16S rDNA technology;To preliminarily explore the possible intervention mechanism of Huayu Wendan Decoction.Methods Ten of 70 male SD rats were randomly selected as blank group,and the remaining 60 rats were fed with high fat and high sugar diet for 21 weeks to induce IGT rat model.30 IGT model rats were randomly divided into model group,Huayu Wendan Decoction group and metformin group,with 10 rats in each group.Huayu Wendan Decoction group was given the TCM decoction for gavage;the metformin group was given the metformin suspension for gavage;the blank group and the model group were given normal saline for gavage.After 4 weeks of drug intervention,FPG and 2 h PG were measured,and feces in the colon were collected.Using 16S rDNA technology,based on OTU clustering results,the diversity and abundance of each group of samples were analyzed,and the species composition of each sample at the phylum and genus level was analyzed.Results The results of Venn diagram based on OTU showed that there were 445 OTUs shared by the four groups,and the similarity among the four groups was high,and the similarity between the blank group and the model group was the lowest.The alpha diversity analysis showed that compared with the blank group,the alpha diversity index of the model group decreased,while the alpha diversity index of Huayu Wendan Decoction group and metformin group increased significantly.The homogeneity and abundance of the microbiome from high to low were the blank group,Huayu Wendan Decoction group,metformin group,and model group.The relative abundance results of species showed that,at the phylum level,Firmicutes was the dominant bacteria in each sample;Saccharibacteria had the largest number in the blank group and the least number in the model group;Euryarchaeota had the largest number in the blank group and almost zero in the model group.At the genus level,the number of Lactobacillus in each sample was the most.The number of Lactobacillus in the blank group was the least,and the number of Lactobacillus in the model group was the most.The analysis of structural differences between the groups showed that comparing the four groups of samples in pairs,the distribution of intestinal flora was obviously different.Conclusion The homeostasis of intestinal flora in IGT rats is unbalanced,and the diversity and abundance of intestinal flora has decreased to a certain extent.Huayu Wendan Decoction may affect the intestinal flora homeostasis of IGT rats by increasing the diversity and relative abundance of gut microbiome.