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  

An Analysis of 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

作　　者：陈吕 张庚鑫 韩雪 唐秋梅 苏钢 杨光勇 陈瑞 何光志 CHEN Lyu;ZHANG Gengxin;HAN Xue;TANG Qiumei;SU Gang;YANG Guangyong;CHEN Rui;HE Guangzhi(Guizhou University of Traditional Chinese Medicine,Guiyang,Guizhou,China,550025;Dejiang People's Hospital,Dejiang,Guizhou,China,565200)

机构地区：[1]贵州中医药大学,贵州贵阳550025 [2]德江县人民医院,贵州德江565200

出　　处：《河南中医》2024年第4期549-555,共7页Henan Traditional Chinese Medicine

基　　金：国家自然科学基金项目(82060796);贵州省科学技术基金项目{黔科合基础[2020]1Y392};贵州省中医药管理局中医药、民族医药科学技术研究课题项目(QZYY-2022-005);贵州省发展和改革委员会工程研究中心建设项目[黔发改高技(2020)896号]。

摘　　要：目的:基于ERIC-PCR技术分析黄连解毒汤对2型糖尿病(diabetes mellitus type 2,T2DM)大鼠肠道菌群结构及DNA同源性的影响。方法:从36只SD大鼠中随机选取6只作为对照组,其余大鼠给予高脂饲料联合1%链脲佐菌素(30 mg·kg^(-1))建立糖尿病模型。将造模成功的大鼠随机分为模型组、黄连解毒汤高剂量组(6.25 g·kg^(-1))、黄连解毒汤中剂量组(3.13 g·kg^(-1))、黄连解毒汤低剂量组(1.56 g·kg^(-1))及二甲双胍组(100 mg·kg^(-1)),每组各6只。各药物组按照相应剂量灌胃给药,对照组和模型组灌胃等体积生理盐水,连续21 d。第7天、第14天及第21天灌胃后,收集大鼠粪便,提取肠道菌群DNA。利用ERIC-PCR技术和Sanger测序检测大鼠肠道菌群结构与DNA同源性。结果:灌胃第7天,对照组条带较多集中在100~750 bp;与对照组比较,模型组条带在1 000~2 000 bp的亮度增强,在300~750 bp的亮度减弱;与模型组比较,黄连解毒汤各剂量组和二甲双胍组条带在750 bp的亮度增强。灌胃第14天,与模型组比较,黄连解毒汤各剂量组和二甲双胍组条带在1 000 bp以上的亮度减弱。灌胃第21天,与模型组比较,黄连解毒汤各剂量组和二甲双胍组条带在300~750 bp的亮度增强。DNA测序结果显示,灌胃第21天,对照组500 bp条带的优势菌群为拟杆菌属、杜氏邓氏菌和金黄杆菌属;模型组400 bp和1 000 bp条带的优势菌群为梭菌目细菌、普拉梭菌、铜绿假单胞菌和杆菌属。灌胃第7天,黄连解毒汤高剂量组750 bp条带的优势菌群为布劳特菌和拟杆菌属;黄连解毒汤中剂量组200 bp条带的优势菌群为脆弱拟杆菌和拟杆菌属;黄连解毒汤低剂量组1 200 bp条带的优势菌群为双发酵副梭菌;二甲双胍组1 000 bp条带的优势菌群为大肠埃希杆菌。灌胃第14天,黄连解毒汤高剂量组700 bp条带的优势菌群为拟杆菌目细菌和假单胞菌;黄连解毒汤中剂量组550 bp条带的优势菌群为假锁杆菌属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 with 1%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 coresponding 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 1000-2000 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 o00 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

关 键 词：黄连解毒汤 2型糖尿病 肠道菌群结构 ERIC-PCR DNA同源性 大鼠 

分 类 号：R285.55[医药卫生—中药学]