基于ERIC-PCR技术分析黄连解毒汤对2型糖尿病大鼠肠道菌群结构及DNA同源性的影响OA

Objective:To analyze the the influence of Coptis Toxin-Resolving Decoction on the structure and DNA homology of intestinal flora in type 2 diabetic rats based on ERIC-PCR technique.Methods:A total of 6 were randomly selected from 36 SD rats as the control group,and the rest rats were given high-fat diet combined with1%streptozotocin(30 mg·kg-1)to establish the diabetes models.The successfully established diabetic rats were randomly divided into the model group,the high-dose group of Coptis Toxin-Resolving Decoc-tion(6.25 g·kg-1),the medium-dose group of Coptis Toxin-Resolving Decoction(3.13 g·kg-1),the low-dose group of Coptis Tox-in-Resolving Decoction(1.56 g·kg-1)and the metformin group(100 mg·kg-1),with 6 rats in each group.The drug groups were given the corresponding dose by gavage;the control group and the model group were given equal volume of saline by gavage for 21 con-secutive days.After gavage on the 7th,14th and 21st day,the rats'feces were collected and intestinal flora DNA was extracted respec-tively.The structure and DNA homology of intestinal flora in rats were detected by ERIC-PCR technique and Sanger sequencing.Re-sults:On the 7th day after gavage,the bands in the control group were mostly concentrated in 100-750 bp.Compared with the control group,the brightness of the model group was enhanced in the 1 000-2 000 bp band and weakened in the 300-750 bp band;compared with the model group,the brightness of the Coptis Toxin-Resolving Decoction dose groups and the metformin group was enhanced in the 750 bp band.On the 14th day after gavage,compared with the model group,the brightness of the Coptis Toxin-Resolving Decoction dose groups and the metformin group was weakened in the 1 000 bp band.On the 21st day after gavage,compared with the model group,the brightness of the Coptis Toxin-Resolving Decoction dose groups and the metformin group was enhanced in the 300-750 bp band.The results of DNA sequencing showed that on the 21st day after gavage,the dominant flora of 500 bp band in the control group was Bacte-roides sp.,Duncaniella dubosii and Chryseobacterium sp.;the dominant flora of 400 bp and 1 000 bp band in the model group was Clos-tridiales bacterium CCNA10,Faecalibacterium prausnitzii,Pseudomonas aeruginosa and Flavonifractor plautii.On the 7th day after ga-vage,the dominant flora of 750 bp band in the high-dose group of Coptis Toxin-Resolving Decoction was Brautia and Bacteroides sp.;the dominant flora of 200 bp band in the medium-dose group of Coptis Toxin-Resolving Decoction was Bacteroides fragilis and Bacteroides sp.;the dominant flora of 1 200 bp band in the low-dose group of Coptis Toxin-Resolving Decoction was Paraclostridium bifermentans;and the dominant flora of 1 000 bp band in the metformin group was Escherichia coil.On the 14th day after gavage,the dominant flora of 700 bp band in the high-dose group of Coptis Toxin-Resolving Decoction was Bacteroidales bacterium and Pseudomonas sp.;the domi-nant flora of 550 bp band in the medium-dose group of Coptis Toxin-Resolving Decoction was Pseudoclavibacter sp.and Streptomyces sp.;the dominant flora of 350 bp band in the low-dose group of Coptis Toxin-Resolving Decoction was Muribaculum gordoncarteri,Muri-baculum intestinale,Duncaniella dubosii and Muribaculaceae bacterium;and the dominant flora of 290 bp band in the metformin group was Escherichia fergusonii and Escherichia coli.On the 21st day of gavage,the dominant bacterial community of the 1 000 bp band in the high-dose group of Coptis Toxin-Resolving Decoction was Prevotella sp.;the dominant bacterial community of the 600 bp band in the medium-dose group of Coptis Toxin-Resolving Decoction was Muribaculum gordoncarteri,Duroc-Deng's bacteria and Bacteroides;the dominant bacterial community of the 400 bp band in the metformin group was Muribaculum gordoncarteri,Enterobacteriaceae,Prevotella sp.,Bacteroides and Bacillus.