化瘀温胆汤对糖耐量受损大鼠肠道菌群多样性及丰度影响

Effect of Huayu Wendan Decoction(化瘀温胆汤)on Diversity and Abundance of Gut Microflora in Impaired Glucose Tolerance Rats

目的通过分析糖耐量受损(IGT)大鼠肠道菌群的差异性,初步探讨化瘀温胆汤对IGT大鼠肠道菌群多样性及丰度的干预情况。方法除空白组(n=10)的SD大鼠,其余SD大鼠予高脂高糖饮食诱导IGT大鼠模型,造模成功后随机分为模型组、化瘀温胆汤组和二甲双胍组(n=10),化瘀温胆汤组给予化瘀温胆汤煎剂1.25 g/mL灌胃,二甲双胍组予5 mg/mL二甲双胍悬浊液灌胃,药物干预4周后采集结肠内粪便。利用QIAamp DNA Stool Mini Kit试剂盒进行粪便样本DNA的提取,分析PCR扩增基因组DNA,构建Illumina平台文库并测序,通过生物信息分析软件对测序结果进行分析,比较各组样本菌群多样性及丰度。结果Rank-Abundance曲线显示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;α多样性指数结果显示与空白组比较,模型组大鼠ACE、Chao1、Shannon、Simpson指数明显降低;经化瘀温胆汤组、二甲双胍组干预后上述指数明显升高;β多样性分析结果显示空白组与模型组各自相对聚集,区分良好,化瘀温胆汤、二甲双胍组虽与空白组、模型组有部分聚集,但整体可以区分开;物种相对丰度结果显示:(1)在门水平下,各样本以Firmicutes(厚壁菌门)为优势菌,Saccharibacteria及Euryarchaeota(广古菌门)在空白组数量最多,模型组数量最少;(2)在属水平下,模型组Lactobacillus(乳酸菌属)数量最多,在空白组数量最少;通过Metastats差异分析显示,在属水平上化瘀温胆汤组Blautia(布劳特氏菌属)、Streptococcus(链球菌属)的相对丰度明显低于模型组。结论IGT大鼠肠道菌群多样性及丰度均降低,化瘀温胆汤可以调节IGT大鼠肠道菌群,降低IGT模型大鼠Blautia(布劳特氏菌属)及Streptococcus(链球菌属)的丰度。

Objective To investigate the effect of Huayu Wendan Decoction(化瘀温胆汤)on the diversity and abundance of gut microbiome in impaired glucose tolerance(IGT)rats by analyzing the difference of gut microbiome in IGT rats.Methods Except for the blank group(n=10),the other SD rats were given high-fat and high sugar diet to induce IGT rat model.After successful modeling,they were randomly divided into model group,Huayu Wendan Decoction group and metformin group(n=10).Huayu Wendan Decoction group was given Huayu Wendan Decoction 1.25 g/mL by gavage,and metformin group was given metformin suspension 5 mg/m L by gavage.The feces in colon were collected after 4 weeks of drug intervention.After 4 weeks of drug intervention,the rats’FPG and 2 h PG were measured,and the colon feces were collected.The DNA of stool samples was extracted by QIAamp DNA Stool Mini Kit.Analyze PCR amplified genomic DNA.The Illumina platform library was constructed and sequenced.The sequencing results were analyzed by bioinformatics analysis software,and the composition and diversity and abundance of bacteria in each group were compared.Results The Rank-Abundance curve showed that the uniformity and abundance of the microbiome from high to low was blank group,Huayu Wendan Decoction group,metformin group,and model group.The alpha diversity index results showed:compare with blank group,the index of ACE,Chao1,Shannon and Simpson were significantly reduced in the model group;compared with the model group,after the intervention of the Huayu Wendan Decoction group and the metformin group,the above indexes increased significantly.The results ofβdiversity analysis showed that the blank group and the model group were relatively clustered and distinguished well.Although the Huayu Wendan Decoction and metformin groups were partially clustered with the blank group and the model group,they could be distinguished as a whole.The relative abundance results of species showed that:(1)At the phylum level,Firmicutes was the dominant bacteria in each sample;Saccharibacteria and Euryarchaeota have the largest number in the blank group and the least number in the model group.(2)At the genus level,the number of Lactobacillus in the model group was the most,while that in the blank group was the least;Metastats difference analysis showed that the relative abundance of Blautia and Streptococcus in the Huayu Wendan Decoction group was significantly lower than the model group at the genus level.Conclusion The diversity and abundance of the gut microbiome of IGT rats are reduced.Huayu Wendan Decoction can regulate the gut microbiome of IGT rats and reduce Blautia and Streptococcus of IGT model rats genus abundance.