高通量测序分析慢性阻塞性肺疾病模型大鼠肠道菌群结构差异及功能  

Analysis of gut microbiome structure and function in a chronic obstructive pulmonary disease model rat using high-throughput sequencing

作　　者：平昕翀 张常喜 马琴 孙雪 PING Xinchong;ZHANG Changxi;MA Qin;SUN Xue(College of Integrated Chinese and Western Medicine,Gansu University of Chinese Medicine,Lanzhou,Gansu,730101,China;Department of Respiratory and Critical Care Medicine,Ningxia Hui Autonomous Region Hospital of Chinese Medicine and Academy of Chinese Medicine,Yinchuan,Ningxia Hui Autonomous Region,750021,China)

机构地区：[1]甘肃中医药大学中西医结合学院,甘肃兰州730101 [2]宁夏回族自治区中医医院暨中医研究院呼吸与危重症医学科,宁夏银川750021

出　　处：《甘肃中医药大学学报》2024年第6期8-16,共9页Journal of Gansu University of Chinese Medicine

基　　金：宁夏回族自治区重点研发项目东西部合作专项(2020BFG03004)。

摘　　要：目的利用16S rRNA基因测序技术探究大鼠肠道菌群差异及其在慢性阻塞性肺疾病(COPD)进程中的潜在作用。方法将10只3月龄SPF级雄性SD大鼠采用完全随机数字表法分为正常组和模型组,每组5只。正常组呼吸新鲜空气联合生理盐水气管滴注,模型组通过烟熏联合脂多糖(LPS)气管滴注法构建COPD大鼠模型,造模7周后采集结肠内粪便进行16S rRNA基因测序及生物信息学分析,比较2组大鼠肠道微生物丰度及多样性变化,并进行差异菌属的鉴定及功能预测。结果与正常组比较,模型组菌群结构科水平丰度下降,属水平丰度提高;物种差异分析显示,模型组和正常组所特有的扩增产物序列变量(ASVs)分别为5545个和5389个,二者相同样本ASVs 633个;线性判别分析效应量(LEfSe)分析显示,模型组与正常组之间差异物种23条,其中棒状杆菌属、放线菌目等在模型组中富集;关联网络分析从门水平对优势物种进行注释,厚壁菌门、拟杆菌门等存在显著聚集并呈正相关,具体到属水平包括罗伊氏乳杆菌属、志贺氏菌属等;京都基因与基因组百科全书(KEGG)功能注释显示,代谢功能模块中氨基酸与碳水化合物代谢等是最大的功能类群。结论COPD模型大鼠肠道内关联炎症活动、肺部基质沉积的菌群表达增加,短链脂肪酸产生菌属丰度下降,肠道菌群或通过影响机体对呼吸系统黏膜免疫的调控而参与COPD疾病进程。Objective To investigate the differences in gut microbiota and its potential role in the progression of chronic obstructive pulmonary disease(COPD)using 16S rRNA gene sequencing technology.Methods 103-month-old SPF male SD rats were randomly divided into a normal group and a model group using a completely random number table,with 5 rats in each group.The normal group was exposed to fresh air and intra-tracheal injection of normal saline,while the model group was exposed to smoke and intra-tracheal injection of lipopolysaccharide(LPS)to establish a COPD rat model.Gut contents were collected from the colon of the model rats 7 weeks after the model was established for 16S rRNA gene sequencing and bioinformatics analysis to compare the changes in gut microbial abundance and diversity between the two groups of rats and identify differential bacterial genera and their functional prediction.Results Compared with the normal group,the abundance of the model group decreased at family level and increased at genus level.Species difference analysis showed that the ASVs specific to the model group and the normal group were 5545 and 5389,respectively,with 633 common ASVs between the two groups.Linear discriminant analysis effect size(LEfSe)analysis showed that there were 23 species that differed between the model group and the normal group,including Bacillus spp.and Actinobacteria orders,which were enriched in the model group.The association network analysis annotated the dominant species at the phylum level,showing significant clustering and positive correlation for Firmicutes and Bacteroidetes,and at the genus level including Lactobacillus and Shigella.The Kyoto Enoyclopedia of Genes and Genomes(KEGG)functional annotation showed that the largest functional group was metabolic functions,including amino acid and carbohydrate metabolism.Conclusion The microbial expression in the intestine of COPD model rats associated with inflammatory activity and pulmonary matrix deposition increased,while the abundance of short-chain fatty ac

关 键 词：慢性阻塞性肺疾病 肠道菌群 16S rRNA基因测序 功能预测 

分 类 号：R563[医药卫生—呼吸系统] R318.04[医药卫生—内科学]