非酒精性脂肪肝与酒精性肝病大鼠肠道菌群比较研究  

Comparison of the gut microbiota in non-alcoholic fatty liver disease and alcoholic liver disease in rats

作　　者：许文琦 王生[1] 黄逸安 王艳 梅其炳[1,2] 刘莉 

机构地区：[1]中国医药工业研究总院上海医药工业研究院创新药物与制药工艺国家重点实验室,上海200437 [2]上海市生物物质成药性评价专业技术服务中心,上海200437

出　　处：《世界临床药物》2018年第1期35-42,共8页World Clinical Drug

基　　金：上海市青年科技杨帆计划项目(编号:17YF1419000)

摘　　要：目的观察非酒精性脂肪肝与酒精性肝病模型大鼠肠道菌群结构变化。方法以高脂饲料(HFD)和酒精液体饲料(LD)分别构建非酒精性脂肪肝和酒精性肝病模型。30只雄性Wistar大鼠随机分成3组,每组10只,按喂养饲料分别分为正常组、HFD组和LD组,喂养6周后解剖取肝脏,检测各组大鼠肝脏病理学改变;收集各组大鼠粪便,提取粪便脱氧核糖核酸进行Illumina MiSeq测序,检测肠道菌群变化。结果肝脏苏木精-伊红染色结果显示,HFD组及LD组大鼠肝脏细胞出现明显脂肪变性病理改变;油红O染色结果显示,HFD组及LD组大鼠肝细胞内大量脂肪沉积。测序结果显示,与正常组相比,HFD组和LD组大鼠菌群多样性明显下降,差异具有统计学意义(P<0.05或P<0.01)。主成分分析结果显示,各组之间群落结构存在明显差异,在门水平上,LD组硬壁菌门相对丰度水平明显高于HFD组(P<0.05);在属水平上,LD组大肠埃希菌-志贺菌属、理研菌科_RC9_gut_组相对丰度低于HFD组,而瘤胃球菌属_1的相对丰度高于HFD组(P<0.05或P<0.01)。组间群落差异分析结果显示,HFD组理研菌科、梭杆菌科、巴斯德氏菌科和气球菌科的相对丰度显著高于LD组,瘤胃球菌科_NK4A214_组、理研菌科_RC9_gut_组、另枝菌属、Marvinbryantia、梭杆菌属、帕匹杆菌属、巴斯德氏菌属、厌氧棍状菌属及格鲁卡比氏菌属等在HFD组中相对丰度较高,而瘤胃球菌属_1、毛螺菌科_NK4A136_组及拟普雷沃菌属在LD组中相对丰度较高。结论非酒精性脂肪肝与酒精性肝病大鼠肠道菌群均处于紊乱状态,且两者存在一定差异。Objective To compare the structural changes of gut microbiota in non-alcoholic fatty liver disease（NAFLD）and alcoholic liver disease（ALD） in rats. Methods Thirty Wistar rats were randomized into three groups（10 rats each group）： normal group, high-fat diet（HFD） group, Lieber-De Carli liquid diet（LD） group. The NAFLD and ALD rat models were established by feeding HFD and LD, respectively. After six weeks, the changes of liver histopathology were assessed in all groups. Their stools were collected and the composition of gut microbiota was determined by Illumina MiSeq sequencing. Results HE staining showed obvious hepatic pathological change and ORO staining showed obvious hepatic fat deposition in HFD group and LD group. The diversity of gut microbiota in HFD group and LD group was significantly decreased compared to normal group（P〈0.05 or P〈0.01）. The PCA showed obvious difference in microbial community structure among three groups. At phylum level, the relative abundance of Firmicutesin in LD group was increased compared to HFD group（P〈0.05）. At genus level, the relative abundance of Escherichia-Shigella and Rikenellaceae_RC9_gut_group was decreased while Ruminococcus_1 was increased in LD group compared to HFD group（P〈0.05 or P〈0.01）. The LEf Se analysis showed that the Rikenellaceae, Fusobacteriaceae, Pasteurellaceae and Aerococcaceaewere were significantly enriched in HFD group compared to LD group. The Ruminococcaceae_NK4 A214_group,Rikenellaceae_RC9_gut_gro up, Alistipes, Marvinbryantia, Fusobacterium, Papillibacter, Pasteurella, Anaerotruncus and Globicatella were the key phylotypes of HFD group while Ruminococcus_1, Lachnospiraceae_NK4 A136_group and Alloprevotella were the key phylotypes of LD group. Conclusion The gut microbiota dysbiosis is present both in NAFLD and ALD in rats, but their gut microbiota is still different significantly.

关 键 词：非酒精性脂肪肝 酒精性肝病 肠道菌群 Illumina测序 

分 类 号：R575.5[医药卫生—消化系统]