二甲双胍对高脂饮食诱导的非酒精性脂肪性肝病大鼠肠道菌群的影响  

Effects of metformin on the intestinal microbiota of rats with high-fat diet-induced non-alcoholic fatty liver disease

作　　者：李瑞芳 卜淑蕊 Li Ruifang;Bu Shurui(Department of Gastroenterology,Jinshan Hospital of Fudan University,Shanghai 201508,China)

机构地区：[1]复旦大学附属金山医院消化内科,上海201508

出　　处：《中华消化杂志》2020年第12期861-867,共7页Chinese Journal of Digestion

摘　　要：目的探讨二甲双胍对非酒精性脂肪性肝病(NAFLD)大鼠肠道菌群的影响。方法雄性Sprague Dawley大鼠24只,适应性喂养2周后,将其分为健康对照组、NAFLD组和二甲双胍治疗组,每组8只。采用高脂饮食诱导建立NAFLD大鼠模型。采集血液标本检测各组ALT、AST、总胆固醇和三酰甘油水平。采用H-E和油红O染色观察肝脏组织病理学变化。根据操作分类单元聚类信息对大鼠肠道菌群结构进行α多样性分析(Chao指数和Shannon指数等)和主坐标分析(PCoA)。采用t检验、单因素方差分析、秩和检验进行统计学分析。结果二甲双胍治疗组大鼠血清ALT、AST、总胆固醇和三酰甘油水平均低于NAFLD组[分别为(58.43±4.89) U/L比(80.14±4.18) U/L、(160.14±13.04) U/L比(203.29±15.11) U/L、(1.534±0.113) mmol/L比(2.003±0.180) mmol/L、(0.724±0.105) mmol/L比(1.030±0.149) mmol/L],差异均有统计学意义(t=-8.926、-5.719、-5.828、-4.446,P均<0.05)。H-E和油红O染色结果显示,二甲双胍治疗后大鼠肝细胞排列较整齐,肝细胞肿胀程度较NAFLD组减轻,脂肪空泡数量减少,肝细胞红染明显减轻,脂滴数量减少,脂滴体积小且分布不均匀。二甲双胍治疗组Chao指数和Shannon指数均低于NAFLD组[690(668,703)比722(683,743)和4.16(4.09,4.42)比4.40(4.20,4.53)],差异均有统计学意义(Z=-2.106、-2.083,P均<0.05)。PCoA结果表明,NAFLD组和二甲双胍治疗组间的整体群落组成不同。在门水平,NAFLD组大鼠肠道菌群中厚壁菌门、变形菌门、放线菌门丰度均高于健康对照组(54.7%比45.5%、3.5%比2.1%、0.2%比0.1%),而拟杆菌门、柔膜菌门丰度均低于健康对照组(41.3%比51.9%、9.4×10^(-7)比2.7×10^(-3)),差异均有统计学意义(Z=-2.866、-2.726、-2.351、-2.901、-4.881,P均<0.05)。在属水平,与健康对照组相比,NAFLD组丰度增加的菌属主要有布劳特氏菌属、拟杆菌属、罗斯氏菌属(0.6%比11.8%、0.9%比6.5%、1.1%比7.1%),而普氏菌属、未�Objective To explore the effects of metformin on the intestinal microbiota of rats with non-alcoholic fatty liver disease(NAFLD).Methods After two weeks of adaptive feeding,24 male Sprague Dawley rats were divided into health control group,NAFLD group and metformin treatment group.NAFLD rat model was induced by high-fat diet.The blood samples were collected and the levels of alanine aminotransferase(ALT),aspartate aminotransferase(AST),total cholesterol(TC)and triglyceride(TG)of each group were detected.The pathological changes of liver tissues were detected by hematoxylin-eosin(H-E)staining and oil red O staining.Theαdiversity analysis,including Chao index and Shannon index,and principal co-ordinates analysis(PCoA)were performed to study the structure of intestinal microbiota of rats according to the clustering information of operational taxonomic unit.T test,one-way analysis of variance and rank sum test were used for statistical analysis.Results The levels of ALT,AST,TC and TG of the metformin treatment group were all lower than those of the NAFLD group((58.43±4.89)U/L vs.(80.14±4.18)U/L,(160.14±13.04)U/L vs.(203.29±15.11)U/L,(1.534±0.113)mmol/L vs.(2.003±0.180)mmol/L,(0.724±0.105)mmol/L vs.(1.030±0.149)mmol/L,respectively),and the differences were statistically significant(t=-8.926,-5.719,-5.828 and-4.446,all P<0.05).The results of H-E staining and oil red O staining showed that after metformin treament,the liver cells of rats were arranged more orderly,the degree of swelling of hepatocytes was reduced,and the number of fat vacuoles decreased,the red staining of hepatocytes was significantly reduced,the number of lipid droplets reduced and lipid droplets were small in size and distributed unevenly in hepatocytes.The Chao and Shannon indexes of the metformin treatment group were both significantly lower than those of the NAFLD group(690(668,703)vs.722(683,743)and 4.16(4.09,4.42)vs.4.40(4.20,4.53)),and the differences were statistical significant(Z=-2.106 and-2.083,both P<0.05).The results of PCoA show

关 键 词：非酒精性脂肪性肝病 高脂饮食 二甲双胍 肠道菌群 16S rDNA测序 

分 类 号：R575.5[医药卫生—消化系统]