MicroRNA-194缓解高脂环境下肝细胞脂质沉积、炎症反应的分子机制研究  

Study on the molecular mechanism of MicroRNA-194 alleviating lipid deposition and inflammation in lipotoxic hepatocytes

作　　者：罗昕 徐梓馨 周璀 徐铭益[1] LUO Xin;XU Zi-xin;ZHOU Cui;XU Ming-yi(Department of Gastroenterology,Shanghai General Hospital,Shanghai Jiao Tong University School of Medicine,Shanghai 200080 China)

机构地区：[1]上海交通大学附属第一人民医院消化内科,200080

出　　处：《肝脏》2021年第4期435-438,450,共5页Chinese Hepatology

基　　金：国家自然科学基金面上项目(81770597);国家科技部十三五项目(2017ZX10203202003005)。

摘　　要：目的研究microRNA-194(miR-194)缓解高脂环境下肝细胞脂质代谢及炎症反应的分子机制。方法以低脂饲料(low-fat diet,LFD)及高脂饲料(high-fat diet,HFD)喂养构建对照组及脂肪肝小鼠模型,HE染色、油红染色观察建模情况。qPCR检测二组小鼠肝组织中miR-194、促脂质合成因子SCD1、ACC、促炎因子TNF-α、IL-6表达水平。棕榈酸(palmitic acid,PA)刺激肝细胞株LO2,过表达miR-194后qPCR检测人肝细胞系LO2细胞内SCD1、ACC、TNF-α、IL-6表达水平,油红染色检测LO2细胞内脂质沉积情况,免疫荧光检测LO2细胞内TNF-α变化。结果脂肪肝小鼠模型构建成功。与LFD组小鼠相比,HFD组小鼠miR-194表达明显降低[LFD:1.000±0.147 vs HFD:0.634±0.116,(t=3.478,P=0.025)]。SCD1、ACC1表达明显升高[SCD1 LFD:1.000±0.287 vs HFD:1.658±0.216,(t=4.802,P=0.009);ACC LFD:1.000±0.252 vs HFD:1.851±0.245,(t=4.194,P=0.015)],TNF-α、IL-6表达明显升高[TNF-αLFD:1.000±0.172 vs HFD:1.952±0.147,(t=7.288,P=0.002);IL-6 LFD:1.000±0.207 vs HFD:1.452±0.108,(t=3.242,P=0.029)]。在高脂环境下,过表达miR-194后,qPCR结果显示LO2细胞内SCD1、ACC表达明显下降[SCD1 miR-NC:1.000±0.149 vs miR-194:0.625±0.112,(t=3.340,P=0.029);ACC miR-NC:1.000±0.204 vs miR-194:0.572±0.124,(t=3.105,P=0.038)],TNF-α、IL-6表达明显减少[TNF-αmiR-NC:1.000±0.149 vs miR-194:0.563±0.059,(t=4.723,P=0.009);IL-6 miR-NC:1.000±0.156 vs miR-194:0.685±0.112,(t=2.853,P=0.048)]。油红染色显示LO2细胞内脂质沉积明显减少,免疫荧光结果显示细胞内TNF-α表达明显下[miR-NC:1.000±0.124 vs miR-194:0.655±0.152,(t=3.020,P=0.039)],炎症反应明显减轻。结论miR-194可以延缓肝细胞内脂质沉积,减轻高脂处理诱导的炎症反应,延缓脂肪肝的进展,这可能成为治疗非酒精性脂肪性肝病的新靶点。Objective To study the molecular mechanism of microRNA-194(miR-194)alleviating lipid metabolism and inflammation in lipotoxic hepatocytes.Methods Low-fat diet(LFD)and high-fat diet(high-fat diet,HFD)were applied to construct a control group and a fatty liver mouse model.HE staining and oil red staining were used to observe the modeling.The expression levels of miR-194,lipid synthesis factors SCD1,ACC,proinflammatory factors TNF-αand IL-6 in the two groups of mice liver tissues were detected by qPCR.Palmitic acid(PA)was used to stimulate the hepatocyte cell line LO2.After miR-194 overexpression in LO2,qPCR was used to detect the expression levels of SCD1,ACC,TNF-α,IL-6 in LO2.Oil red staining was used to detect the lipid deposition in LO2,and immunofluorescence was performed to analyze the changes of TNF-αin LO2.Results HE and oil red staining showed that the fatty liver mouse model was successfully constructed.Compared with the mice in the LFD group,the expression of miR-194 in the HFD group was significantly reduced(LFD:1.000±0.147 vs HFD:0.634±0.116,t=3.478,P=0.025).The expression of SCD1 and ACC1 were significantly increased(SCD1 LFD:1.000±0.287 vs HFD:1.658±0.216,t=4.802,P=0.009;ACC LFD:1.000±0.252 vs HFD:1.851±0.245,t=4.194,P=0.015),and the expression of TNF-αand IL-6 were also greatly promoted(TNF-αLFD:1.000±0.172 vs HFD:1.952±0.147,t=7.288,P=0.002;IL-6 LFD:1.000±0.207 vs HFD:1.452±0.108,t=3.242,P=0.029).In the high-fat environment,after overexpression of miR-194,qPCR results showed that the expression of SCD1 and ACC in LO2 decreased significantly(SCD1 miR-NC:1.000±0.149 vs miR-194:0.625±0.112,t=3.340,P=0.029;ACC miR-NC:1.000±0.204 vs miR-194:0.572±0.124,t=3.105,P=0.038),and the expression of TNF-αand IL-6 decreased significantly(TNF-αmiR-NC:1.000±0.149 vs miR-194:0.563±0.059,t=4.723,P=0.009;IL-6 miR-NC:1.000±0.156 vs miR-194:0.685±0.112,t=2.853,P=0.048).Oil red staining showed that the lipid deposition in LO2 was significantly alleviated,and the immunofluorescence results showed t

关 键 词：微小RNA-194 脂质沉积 炎症反应 非酒精性脂肪性肝病 

分 类 号：R57[医药卫生—消化系统]