过表达miR-140-5p对高脂饲养诱导的非酒精性脂肪性肝病大鼠的作用及机制  

Effect and mechanism of miR-140-5p overexpression on rats with non-alcoholic fatty liver disease induced by high-fat feeding

作　　者：卢的一 郭茂东 叶晓华[1] LU Deyi;GUO Maodong;YE Xiaohua(Department of Gastroenterology,Jinhua Central Hospital,Jinhua 321000,China)

机构地区：[1]金华市中心医院消化内科,321000

出　　处：《国际消化病杂志》2021年第1期28-32,共5页International Journal of Digestive Diseases

摘　　要：目的探究过表达miR-140-5p对高脂饲养诱导的非酒精性脂肪性肝病(NAFLD)大鼠的作用及机制。方法40只大鼠随机分为对照组、模型组、空载体组和过表达组,每组各10只。对照组给予普通饲料饲养,模型组给予高脂饲料饲养构建NAFLD大鼠模型,空载体组在模型组基础上于饲养第13周、第18周尾静脉注射含空载质粒的慢病毒,过表达组在模型组基础上于饲养第13周、第18周尾静脉注射含过表达miR-140-5p质粒的慢病毒。采用酶联免疫吸附法检测各组血清天冬氨酸氨基转移酶(AST)、丙氨酸氨基转移酶(ALT)、总胆固醇(TC)、三酰甘油(TG)、低密度脂蛋白胆固醇(LDL-C)、白细胞介素-6(IL-6)和肿瘤坏死因子-α(TNF-α)的表达水平。采用蛋白质免疫印迹法检测各组肝脏组织中Toll样受体4(TLR4)、核因子-κB(NF-κB)的表达水平。采用实时荧光定量PCR法检测miR-140-5p的相对表达量。结果与对照组比较,模型组miR-140-5p的相对表达量降低,而AST、ALT、TC、TG、LDL-C、IL-6、TNF-α、TLR4和NF-κB水平升高,差异均有统计学意义(P均<0.05)。与模型组和空载体组比较,过表达组miR-140-5p的相对表达量升高,AST、ALT、TC、TG、LDL-C、IL-6、TNF-α、TLR4和NF-κB水平降低,差异均有统计学意义(P均<0.05)。结论过表达miR-140-5p可改善NAFLD大鼠的肝功能,纠正脂代谢紊乱,其机制可能与调控TLR4/NF-κB信号通路有关。Objective This paper aims to investigate the effect and mechanism of miR-140-5p overexpression on rats with non-alcoholic fatty liver disease(NAFLD)induced by high-fat feeding.Methods Forty rats were randomly assigned to the control group,the model group,the empty vector group,and the overexpressed group,with 10 rats in each group.The control group was fed with ordinary diet,the model group was fed with high-fat diet to construct the NAFLD rat model,while the empty vector group was injected with lentivirus containing empty plasmid through tail vein at the 13th and 18th week based on the model group,and the overexpressed group was injected with lentivirus containing the overexpressed miR-140-5p plasmid through tail vein at the 13th and 18th week based on the model group.The levels of aspartate aminotransferase(AST),alanine aminotransferase(ALT),total cholesterol(TC),triglyceride(TG),low-density lipoprotein cholesterol(LDL-C),interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α)were detected by enzyme-linked immunosorbent assay.The toll like receptor 4(TLR4)and nuclear factorκB(NF-κB)in the liver were detected by Western blot.The levels of miR-140-5p were detected by real-time fluorescence quantitative PCR.Results Compared with the control group,the relative expression of miR-140-5p in the model group was reduced,and the levels of AST,ALT,TC,TG,LDL-C,IL-6,TNF-α,TLR4,and NF-κB were increased(P<0.05).Compared with the model group and the empty vector group,the relative expression of miR-140-5p was increased in the overexpression group,and the levels of AST,ALT,TC,TG,LDL-C,IL-6,TNF-α,TLR4,and NF-κB were reduced,with statistically significant differences(P<0.05).Conclusions Overexpression of miR-140-5p can improve liver function and correct lipid metabolism disorders in NAFLD rats.The mechanism may be related to the regulation of TLR4 and NF-κB expression.

关 键 词：非酒精性脂肪性肝病 miR-140-5p TOLL样受体4 核因子-ΚB 

分 类 号：R575.5[医药卫生—消化系统]