高通量测序研究中药经方对正常小鼠肠道菌群的影响

Effects of Traditional Chinese Medicine Prescriptions on the Intestinal Flora of Normal Mice via High-throughput Sequencing

目的:探讨四君子汤、麻杏石甘汤、麻杏石甘汤合四君子汤对正常小鼠肠道菌群多样性的影响。方法:将20只Balb/c小鼠随机分为正常组、四君子汤组、麻杏石甘汤组、麻杏石甘汤合四君子汤组4组,每组5只,除正常组给予无菌双蒸水灌胃外,其余3组分别以对应汤药灌胃。于灌胃第4、7天无菌收集各组小鼠粪便,16S rRNA高通量测序法检测各组药物对小鼠粪便中微生物群落结构及多样性变化的调节。结果:各药物的干预使正常小鼠肠道菌群的OTU(operational taxonomic unit)数量产生了变化,麻杏组菌群丰富度增加尤为显著( P <0.05);2时间段各中药组的菌群香农指数(Shannon index)较正常组比较均有变化、麻杏组的菌群多样性增加更为明显;正常组分别与给药组比较,正常组与四君组间共有的OTUs数目最多,其菌群间的相似性最高;药物的使用会使小鼠肠道的优势菌群发生结构及比例的变化;给药组与正常组小鼠的总体菌群在结构上分离明显、差异有统计学意义;四君子汤、麻杏石甘汤合四君子汤的使用会显著性地使与短链脂肪酸(SCFA)产生相关的菌属丰度增加。结论:与正常组比较,各中药组小鼠肠道菌群结构及多样性均发生明显变化。

Objective: To explore the effects of Sijunzi Decoction,Maxingshigan Decoction,the combination of Maxingshigan and Sijunzi Decoction on the diversity of intestinal flora in normal mice. Methods: A total of 20 Balb/c mice were randomly divided into 4 groups:the normal group,the Sijunzi Decoction group,the Maxingshigan Decoction group,the combined Maxingshigan and Sijunzi Decoction group,with 5 mice in each group.Sterile double distilled water was administrated to the normal group by gavage,while the other 3 groups were given the corresponding decoction by gavage.The feces of mice in each group were collected aseptically on the 4th and 7th day of gavage.The 16S rRNA high-throughput sequencing method was used to detect the changes of microbial community structure and diversity in the feces of mice in each group. Results: The intervention of each drug resulted in changes in the numbers of OTU(operational taxonomic unit)in the intestinal flora of normal mice,and the increase in the abundance of the Maxingshigan Decoction group was especially significant( P <0.05).In the two time periods,the Shannon indexes of all traditional Chinese medicine groups were different from that of the normal group,and the increase of the flora diversity of the Mmaxingshigan Decoction group was more significant.Compared individually with each drug group,the number of OTUs shared between the normal group and the Sijunzi Decoction group was the highest,and the similarity of the two intestinal floras was the highest.The use of drugs changed the structure and proportion of dominant bacteria in the intestinal tract of mice.The total microflora of mice in the drug groups and the normal group were significantly isolated from each other.The use of Sijunzi Decoction and the combination of Maxingshigan and Sijunzi Decoction can significantly increase the abundance of bacteria related to SCFA production. Conclusion: Compared with the normal group,the structure and diversity of intestinal flora of mice in every Chinese medicine group changed significantly.