miR-490-3p过表达对大鼠非酒精性脂肪性肝病的改善作用及其机制  

Improvement effect of miR-490-3p over-expression on non-alcoholic fatty liver disease in rats and its mechanism

作　　者：龚杰 雷泽华[1] 张远威 黄雄[3] 杜波[1] 王志旭[1] GONG Jie;LEI Zehua;ZHANG Yuanwei;HUNANG Xiong;DU Bo;WANG Zhixu(Department of Hepatopancreatobiliary Surgery,Leshan People’s Hospital,Leshan Treatment Center of Hepatopancreatobiliary System Diseases,Sichuan Province,Leshan 614000,China;Department of Gastrointestinal Surgery,Second Affiliated Hospital,Zunyi Medical University,Zunyi 563000,China;Department of General Surgery,Chengdu Sixth People’s Hospital,Sichuan Province,Chengdu 610051,China)

机构地区：[1]四川省乐山市人民医院肝胆胰外科四川省乐山市肝胆胰系统疾病诊疗中心,四川乐山614000 [2]遵义医科大学第二附属医院胃肠外科,贵州遵义563000 [3]四川省成都市第六人民医院普外科,四川成都610051

出　　处：《吉林大学学报（医学版）》2021年第6期1495-1501,共7页Journal of Jilin University：Medicine Edition

基　　金：四川省卫健委科研基金项目(20PJ296)。

摘　　要：目的:探讨miR-490-3p过表达对大鼠非酒精性脂肪性肝病(NAFLD)的改善作用,并初步分析其作用机制。方法:32只雄性SD大鼠随机分为对照组、模型组、阴性对照组(mimic-NC组)和miR-490-3p模拟物组(miR-490-3p mimic组),每组8只。采用高脂饲料喂养复制NAFLD大鼠模型,对照组大鼠不进行任何处理,mimic-NC组和miR-490-3p mimic组大鼠分别尾静脉注射含无意义序列和miR-490-3p mimic质粒的腺病毒。采用实时荧光定量PCR(RT-qPCR)法检测各组大鼠肝组织中miR-490-3p表达水平,HE染色和油红O染色观察各组大鼠肝组织病理形态表现,ELISA法检测各组大鼠血清丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)、肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6)、白细胞介素1β(IL-1β)、丙二醛(MDA)、超氧化物歧化酶(SOD)和谷胱甘肽过氧化物酶(GSH-Px)水平,RT-qPCR法检测各组大鼠肝组织中Toll样受体4(TLR4)和核因子κB(NF-κB)mRNA表达水平,Western blotting法检测各组大鼠肝组织中TLR4和NF-κB蛋白表达水平。结果:与对照组比较,模型组大鼠血清ALT、AST、TNF-α、IL-6、IL-1β和MDA水平和肝组织中TLR4 mRNA及蛋白、NF-κB mRNA表达水平均升高(P<0.05),血清SOD和GSH-Px水平及肝组织中miR-490-3p表达水平均降低(P<0.05);与模型组比较,miR-490-3p mimic组大鼠血清ALT、AST、TNF-α、IL-6、IL-1β、MDA水平和肝组织中TLR4 mRNA及蛋白、NF-κB mRNA表达水平均降低(P<0.05),血清SOD和GSH-Px水平和肝组织中miR-490-3p表达水平升高(P<0.05)。HE染色,对照组大鼠肝细胞排列均匀;模型组和mimic-NC组大鼠肝细胞中堆积大量脂肪,门管区伴有明显炎性细胞浸润,部分肝细胞呈坏死,并出现纤维化病变;miR-490-3p mimic组大鼠肝组织病变程度较模型组减轻。油红O染色,对照组大鼠肝组织中未见明显红染脂肪滴;模型组和mimic-NC组大鼠肝组织中可见明显红染的脂肪滴,数量多且体积大;miR-490-3p mimic组大鼠肝Objective:To explore the improvement effect of miR-490-3p over-expression on nonalcoholic fatty liver disease(NAFLD)of the rats,and to preliminarily analyze its action mechanism.Methods:A total of 32 male SD rats were randomly divided into control group,model group,negative control group(mimic-NC group),and miR-490-3p mimic group(miR-490-3p mimic group),and there were8 rats in each group.The rats were fed with high-fat diet to replicate the NAFLD rat models.The rats in control group were given nothing,and the rats in mimic-NC and miR-490-3p mimic groups were injected with adenovirus containing meaningless sequence and miR-490-3p mimic plasmid through tail vein,respectively.The expression levels of miR-490-3p in liver tissue of the rats in various groups were detected by Real-time fluorescence quantitative(RT-qPCR)method.The serum levels of alanine aminotransferase(ALT),aspartate aminotransferase(AST),tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),interleukin-1β(IL-1β),malondialdehyde(MDA),superoxide dismutase(SOD),glutathione peroxidase(GSH-Px)in liver tissue of the rats in various groups were detected by ELASA method.The pathomorphology of liver tissue of the rats in various groups were detected by HE staining and Oil red O staining.The expression levels of Toll-like receptor 4(TLR4)and nuclear factorκB(NF-κB)mRNA in liver tissue of the rats in various groups were detected by RT-qPCR method.The expression levels of TLR4 and NF-κB proteins in liver tissue of the rats in various groups were detected by Western blotting method.Results:Compared with control group,the serum levels of ALT,AST,TNF-α,IL-6,IL-1βand MDA,the expression levels of TLR4 mRNA and protein and NF-κB mRNA in liver tissue of the rats in model group were increased(P<0.05);while the serum levels of SOD and GSH-Px and expression level of miR-490-3p in liver tissue were decreased(P<0.05).Compared with model group,the serum levels of ALT,AST,TNF-α,IL-6,IL-1β,and MDA and the expression levels of TLR4 mRNA and protein and NF-κB mRNA in l

关 键 词：miR-490-3p 非酒精性脂肪性肝病 炎症反应 氧化应激 Toll样受体4/核因子κB信号通路 

分 类 号：R575.5[医药卫生—消化系统]