糖肾灌肠方经肠道干预糖尿病肾病小鼠模型的肠道菌群研究

Study on Tangshen Guanchang Prescription(糖肾灌肠方)Intervening Intestinal Flora via Intestine of Diabetic Kidney Disease Mice

目的观察中药经肠道干预糖尿病肾病(DKD)小鼠肠道菌群的多样性。方法将72只小鼠建立DKD动物模型后随机分成模型组、糖肾灌肠方组、替米沙坦组、替米沙坦+糖肾灌肠方组和预防组,每组15只,另取12只小鼠做正常组,各组小鼠给予相应的28天治疗后收集新鲜粪便,用Illumina-MiSeq高通量测序技术分析微生物群落结构和组成的变化。结果通过Alpha多样性中Simpson指数整体结果显示,正常组、模型组、糖肾灌肠方组和替米沙坦组之间差异无统计学意义(P>0.05);替米沙坦+糖肾灌肠方组和预防组与其他各组之间有差异有统计学意义(P<0.05)。从门水平上,相对丰度大于5%的门包括:Firmicutes(厚壁菌门,43.9%),Bacteroidetes(拟杆菌门,41.0%),Proteobacteria(变形菌门,6.0%)。这三个菌门总共占到总体菌落的90.9%。其中占比最多的是厚壁菌门,其次是拟杆菌门和变形菌门。从属水平上,Lactobacillus(乳酸菌属,27.0%),Clostridiales(梭菌目属,7.1%),Akkermansia(阿克曼菌属,5.3%),Prevotella(普氏菌属,4.1%),Allobaculum(3.2%),Bacteroides(拟杆菌属,3.1%)占总体的72%。通过QIIME软件,结果显示经药物干预后,乳酸菌属、梭菌目属、普氏菌属、双歧杆菌属明显上升。结论糖肾灌肠方可能通过增加DKD小鼠肠道菌群多样性,影响了肠道菌群的结构,从而对DKD小鼠模型的肾脏起保护作用。

Objective To observe the diversity of intestinal flora in mice with diabetic kidney disease(DKD)treated with traditional Chinese medicine.Methods After establishing DKD animal model,72 mice were randomly divided into model group,Tangshen Guanchang Prescription enema group,telmisartan group,Tangshen Guanchang Prescription enema+telmisartan group and prevention group,with 15 mice in each group.Another 12 mice were taken as the normal group.Fresh feces were collected after 28 days of treatment,and the changes of microbial community structure and composition were analyzed by Illumina miseq high-throughput sequencing technology.Results The overall results of Simpson index in alpha diversity showed that there was no significant difference among normal group,model group,Tangshen Guanchang Prescription enema group and telmisartan group(P>0.05).There was significant difference between Tangshen Guanchang Prescription enema group+telmisartan group and between prevention group and other groups(P<0.05).At the phylum level,phyla with relative abundance greater than 5%included Firmicutes(43.9%),Bacteroidetes(41.0%)and Proteobacteria(6.0%).These three phyla accounted for 90.9%of the total colonies.Among them,Firmicutes accounted for the largest proportion,followed by Bacteroidetes and Proteus.At the subordinate level,Lactobacillus(lactic acid bacteria,27.0%),clostridiales(clostridia,7.1%),akkermansia(akman bacteria,5.3%),Prevotella(Prevotella,4.1%),allobaculum(3.2%)and Bacteroides(Bacteroides,3.1%)accounted for 72%of the total.Through QIIME software,the results showed that after drug intervention,lactic acid bacteria,Clostridium,Prevotella and Bifidobacterium increased significantly.Conclusion The decoction may affect the structure of intestinal flora by increasing the diversity of intestinal flora in DKD mice,and thus protecting the kidney in DKD mice model.