高通量测序技术研究糖尿病大鼠与正常大鼠粪便菌群的结构差异

Comparision of Faecal Microbiota in Rats with Type 2 Diabetes and Non-Diabetic Rats Using Miseq High-throughput Sequencing

为研究糖尿病大鼠和正常大鼠粪便菌群的结构差异,选取SD雄性大鼠,随机分为正常组和糖尿病组,于第4周末,对大鼠粪便菌群16S r RNA的V1~V3区进行扩增,Miseq高通量测序平台测定其基因序列,与silva数据库中微生物基因序列进行比对、分类。研究发现:糖尿病大鼠体重较正常组显著下降;糖尿病大鼠粪便菌群多样性高于正常大鼠;糖尿病大鼠粪便菌群中肠杆菌目,假单孢杆菌目等机会致病菌增多;同时克里斯滕森菌科,明串珠菌科,黄杆菌科,Clostridium sensu stricto 1,Gastranaerophilales_norank,Ruminococcaceae_incertae_sedis等在糖尿病组粪便中大量存在。结果表明:糖尿病大鼠肠道菌群结构与正常大鼠差异显著,存在肠道菌群紊乱的状况,提示临床上可通过调节饮食结构来改善肠道菌群,达到防治2型糖尿病的目的。

The objective of this study was to characterize the composition of fecal microbiota in rats with diabetes type 2 as compared to non-diabetic controls using Miseq high-throughput sequencing of the V1 ~V3 region of the 16 S r RNA gene. Male SD rats were randomly divided to diabetic and non-diabetic groups. The feces were collected after the4-week experimental period. The fecal bacterial composition was investigated by Miseq high-throughput sequencing of the V1~V3region of the 16 S r RNA gene. The results showed that during the 4-week feeding course, the body weight of diabetic rats was significantly reduced compared to non-diabetic rats. Bacterial diversity and richness in diabetic subjects was somewhat higher than in non-diabetic rats. Using the taxonomic characterization from the gene sequence, we found that most of T2D-enriched microbials were from opportunistic pathogens, such as Enterobacteriales, Pseudomonadales.This study also found some microbials enriched in diabetic rats, such as Christensenellaceae, Leuconostocaceae,Flavobacteriaceae, Clostridiumsensustricto1, Gastranaerophilalesnorank, Ruminococcaceaeincertaesedis. Our findings indicated that T2 D rats had a moderate degree gut bacterial dysbiosis, with a decline in beneficial bacteria, and an increase in several opportunistic pathogens.