基于16S rDNA测序技术探讨参苓白术散对溃疡性结肠炎大鼠肠道菌群的影响  

Effect of Shenling Baizhu Powder on Intestinal Flora in Rats with Ulcerative Colitis Based on 16S rDNA Sequencing

作　　者：徐霞 王玮玮 涂钰 张文彤 李冰涛[1,3] 张启云 刘红宁[1,3,4] 徐国良 姜丽[1,3,4] XU Xia;WANG Wei-wei;Tu Yu;ZHANG Wen-tong;LI Bing-tao;ZHANG Qi-yun;LIU Hong-ning;XU Guo-liang;and JIANG Li(Institute of Research Center for Differentiation and Development of CM Basic Theory,Jiangxi University of Chinese Medicine,Nanchang,330004;Department of Pharmacy,Affiliated Hospital of Jinggangshan University,Jiangxi,343000;Jiangxi Provincial Key laboratory of CM Etiopathogenisis,Nanchang,330004;Key Laboratory of Pharmacology of Traditional Chinese Medicine in Jiangxi,Nanchang,330004)

机构地区：[1]江西中医药大学中医基础理论分化发展研究中心,南昌330004 [2]井冈山大学附属医院药剂科,江西343000 [3]江西省中医病因生物学重点实验室,南昌330004 [4]江西省中药药理学重点实验室,南昌330004

出　　处：《中国中西医结合杂志》2023年第11期1334-1342,共9页Chinese Journal of Integrated Traditional and Western Medicine

基　　金：江西省中医药管理局科技计划(No.2021B599);江西省教育厅科学技术研究项目(No.GJJ221256);第四批中青年骨干人才计划;(No.赣中医药科教字[2022]6号);江西中医药大学校级科技创新团队发展计划(No.CXTD22007)。

摘　　要：目的探讨参苓白术散通过调节肠道微生物的组成改善溃疡性结肠炎(UC)的作用及其机制。方法采用连续灌胃5%聚糖硫酸钠(DSS)制备UC大鼠模型,将造模成功的12只大鼠按随机数字表法分为模型组(n=6)、参苓白术散组(n=6),另设正常对照组(n=7)。正常对照组、模型组大鼠灌胃灭菌纯净水(2 mL/d),参苓白术散组灌胃去离子水超声溶解的参苓白术散,剂量为每千克大鼠1.89 g/d,药液浓度为0.125 g/mL,给药30天。观察大鼠的体重、疾病活动指数(DAI)变化,检测各组大鼠血清中的C反应蛋白(CRP)、血清降钙素(PCT)以及诱导型一氧化氮合成酶(iNOS)水平,并通过16S rDNA技术探讨大鼠经参苓白术散干预后肠道微生物结构的变化。结果与正常对照组比较,模型组体重逐渐下降(P<0.05),iNOS水平亦下降(P<0.01),CRP、PCT水平及DAI评分升高(P<0.01);与模型组比较,参苓白术散组大鼠血清CRP、PCT水平下降(P<0.05,P<0.01)。Alpha多样性指数显示:与正常对照组比较,UC模型组Shannon、Chao指数升高(P<0.01,P<0.05),Simpson指数降低(P<0.01)。与模型组比较,参苓白术散组Shannon、Ace指数均降低(P<0.05)。Beta多样性分析中主成分分析(PCA)、主坐标分析(PCoA)、非度量多维尺度分析(NMDS)的结果显示各组内样本相对聚集,各组间样本距离较远,通过Venn图发现了正常对照组、模型组、参苓白术散组中分别特有81、83、73个OTUs。肠道菌群结构分析结果显示:与正常组比较,模型组放线菌门相对丰度及乳酸杆菌降低(P<0.05),变形菌门、拟杆菌门相对丰度增加(P<0.05),脱硫弧菌属、瘤胃梭菌属-9、拟杆菌属升高(P<0.01);与模型组比较,参苓白术散组厚壁菌门丰度增加(P<0.05),拟杆菌门、软壁菌门丰度减少(P<0.05)。物种差异分析结果显示,在门到属水平上,正常对照组中区别于其他组的特征性差异菌有21个,参苓白术散组中区别于其他组的特征性差异菌有10个。最�Objective To explore the effect and mechanism of Shenling Baizhu Powder(SLBZP)on improving ulcerative colitis(UC)by regulating the composition of intestinal microorganisms.Methods The UC rat model was established by continuous gavage of 5%sodium glycan sulfate(DDS).Totally 12 successfully modeled rats were randomly divided into the model group(n=6)and the SLBZP group(n=6).A normal control group consisting of 7 rats were also set up.Rats in the normal control group and model group were administered with sterilized pure water(2 mL·d-1)by gastrogavage,while those in the SLBZP group administered with ultrasound dissolved SLBZP in deionized water by gastrogavage.The dose was 1.89 g·d-1 per kg of rats,and the drug concentration was 0.125 g·mL-1 for 30 days.The changes of body weight and disease activity index of rats were observe.The C-reactive protein(CRP),procalcitonin(PCT),and inducible nitric oxide synthase(iNOS)of rat serum were determine,and to explore the changes of intestinal microbial structure of rats after the intervention of SLBZP through 16S rDNA technology.Results Compared with the normal control group,the weight of the rats gradually decreased(P<0.05),the iNOS also decreased(P<0.01),and the CRP,PCT levels,and DAI scores increased in the model group(P<0.01).Compared with the model group,the serum CRP and PCT levels decreased in the SLBZP group(P<0.05,P<0.01).The Alpha diversity index showed that compared with the normal control group,the Shannon and Chao indices of the UC model group increased(P<0.01,P<0.05),while the Simpson indices decreased(P<0.01).Compared with the model group,the Shannon and Ace indices of the SLBZP group decreased(P<0.05).The Principal Component Analysis(PCA),Principal CO-ordinates Analysis(PCoA),and Non-metric Multidimensional Scaling(NMDS)results in the Beta diversity analysis showed that the samples within each group were relatively clustered,and the distance between the samples was relatively long.Through Venn plots,it was found that there were 81,83,and 73 unique OTUs in the

关 键 词：参苓白术散 16S rDNA 溃疡性结肠炎 肠道菌群 大鼠 中药复方 

分 类 号：R285.5[医药卫生—中药学]