基于16S rRNA基因测序的黄连对2型糖尿病大鼠肠道微生物多样性影响研究

目的探索黄连提取物对2型糖尿病模型大鼠肠道微生物多样性的影响,探讨黄连治疗消渴证的机制。方法随机选取10只SD大鼠为对照组,其余大鼠采用特殊膳食诱导与ip链脲佐菌素(STZ)联合制备2型糖尿病模型,造模成功大鼠随机分为3组:模型组、黄连提取物(4.36 g/kg)组、二甲双胍(0.097 g/kg)组,每组10只;各组均ig给药,对照组与模型组ig等体积的0.5%羟甲基纤维素钠(CMC-Na),每周监测体质量、血糖值各1次,给药4周后测定口服糖耐量以及胰岛素水平,给药4周后取其结肠内容物,进行16S rRNA基因测序。结果黄连提取物对2型糖尿病大鼠症状均有改善作用,16S rRNA基因得到了951个OTU,15个门、25个纲、43个目、69个科、182个属、357个种;门水平,模型组放线菌门(Actinobacteria)细菌的量明显高于对照组与黄连提取物组(P0.05);脱铁杆菌门(Deferribacteres)只在模型组中检出;厚壁菌门(Firmicutes)、螺旋体门(Spirochaetae)、柔膜菌门(Tenericutes)、迷踪菌门(Elusimicrobia)在模型组中的量均高于对照组,而给予黄连提取物后,丰度有所降低;同时黄连提取物组中变形菌门(Proteobacteria)、梭杆菌门(Fusobacteria)、疣微菌门(Verrucomicrobia)的丰度略高于对照组与模型组。结论经膳食联合STZ诱导的2型糖尿病大鼠中放线菌门与脱铁杆菌门可能是潜在的标志菌;给予黄连提取物后,肠道菌群多样性发生改变,提示黄连提取物可以改善2型糖尿病大鼠肠道菌群紊乱。

Based on sequencing of 16 S rRNA gene elaborate effect of Coptis chinensis on intestinal microbial diversity in type 2 diabetes mellitus

Objective To explore the effect of Coptis chinensis on the intestinal microflora diversity of rats with type 2 diabetes mellitus and further elucidate the mechanism of C. chinensis in treatment of type 2 diabetes mellitus. Methods SD rats were randomly divided into control group (CON), model group (M), C. chinensis group (HL), and positive control group (POS), 10 in each group respectively. Type 2 diabetes mellitus model was caused by special diet fed and ip injection of STZ together. CON with M, HL, and POS was respectively given gavage volume 0.5% CMC-Na, 4.36 g/kg C. chinensis water extract and 0.097 g/kg of metformin. The body weight and blood glucose were monitored once a week separately. After four weeks of administration, the oral glucose tolerance and insulin level were measured. After four weeks of administration, the contents of rectum were collected and sequenced by a 16 S rRNA gene sequencing Results A total of 951OTU, 15 Phylum, 25 Class, 43 Order, 69 Family, 182 Genus and 357 Species were obtained. At the level of Phylum, the content of Actinobacteria in M was significantly higher than that in CON and HL (P<0.05), while Deferribacteres was only detected in M. In M, Firmicutes, Spirochaetae, Tenericutes, and Elusimicrobia were higher than CON. Interestingly, they were all reduced after giving C. chinensis, meanwhile the abundance of Proteobacteria, Fusobacteria and Verrucomicrobia in HL were slightly higher than that in CON and M. Conclusion Actinobacteria and Deferribacteres were probably potential markers in type 2 diabetes mellitus. After administrating C. chinensis, the change of intestinal microflora diversity revealed C. chinensis could improve the intestinal microflora disorder of type 2 diabetes mellitus.