miR-22靶向SIRT1促进人脂肪肝细胞中脂肪沉积的机制研究  

The mechanism of miR-22 targeting SIRT1 to promote fat deposition in the human steatotic hepatocyte model

作　　者：郑晓筠 黄浩辉 何杰[1] 李瑜元[1] 聂玉强[1] 杜艳蕾[1] 周永健[1] ZHENG Xiaojun;HUANG Haohui;HE Jie;LI Yuyuan;NIE Yuqiang;DU Yanlei;ZHOU Yongjian(Guangzhou Digestive Disease Center,Guangzhou First People’s Hospital,Guangzhou Medical University,Guangzhou 510180,China)

机构地区：[1]广州医科大学附属广州市第一人民医院广州消化疾病中心,广东广州510180

出　　处：《胃肠病学和肝病学杂志》2018年第2期182-187,共6页Chinese Journal of Gastroenterology and Hepatology

摘　　要：目的探讨正常人肝细胞株L02肝脂肪变时miR-22的表达变化及其靶向SIRT1促进脂肪沉积的功能机制研究。方法用油酸和棕榈酸混合物诱导法建立L02脂肪变肝细胞株模型,验证建模成功后,PCR检测miR-22的表达变化,Western blotting检测SIRT1蛋白表达变化;生物学预测miR-22靶基因;过表达miR-22和抑制miR-22表达,观察油红O染色、油红脱色含量测定、甘油三酯和肝酶比,Western blotting检测SIRT1蛋白表达变化。结果成功建立脂肪肝细胞模型,miR-22在脂肪肝细胞表达显著增加(P<0.05);SIRT1在脂肪肝细胞中蛋白表达显著减少(P<0.05);生物学预测SIRT1是miR-22的靶基因;过表达miR-22后脂滴明显变大、增加,TG含量和AST/ALT显著增大(P<0.05),SIRT1表达减少(P<0.05),抑制miR-22后脂滴明显减少,TG含量显著减少(P<0.05),SIRT1蛋白表达呈增加趋势(P>0.05)。结论 miR-22在肝细胞株脂肪变时表达显著增加,miR-22表达上调负调控SIRT1的表达,促进脂肪性肝病中脂肪沉积。Objective To investigate the expression of miR-22 in the human steatotic hepatocyte model and its function and mechanism of targeting SIRT1 to promote fat deposition.Methods Human hepatic cell line L02 steatosis hepatocyte model was induced with oleic acid and palmitic acid.After successful validation model,the expression of miR-22 was measured by qRT-PCR while the expression of SIRT1 protein was measured by Western blotting.Subsequently,miR-22 mimic and miR-22 inhibitor were transfected into steatotic hepatocytes,the results of oil red O staining,oil red decolorization determination,triglyceride lipid content and liver enzyme ratio were observed and the expression of SIRT1 protein was detected by Western blotting.Results The steatosis hepatocyte model was successfully established.The expression of miR-22 in steatotic hepatocytes was significantly higher than that in the control group(P<0.05),while the expression of SIRT1 protein was significantly decreased(P<0.05).SIRT1 was the predicted target gene for miR-22.After transfection,fat drops were obviously larger and increased in the miR-22 mimic group than those in the control group,its triglyceride lipid and AST/ALT ratio were significantly higher(P<0.05),while the expression of SIRT1 protein was significantly lower(P<0.05).Nevertheless,fat drops were obviously smaller and decreased in the miR-22 inhibitor group than those in the control group,its triglyceride lipid and AST/ALT ratio were significantly lower(P<0.05).The expression of SIRT1 protein was significantly lower in the miR-22 mimic group than that in the control group(P<0.05),while the expression of SIRT1 protein tended to increase in the miR-22 inhibitor group(P>0.05).Conclusion The miR-22 is up-regulated in steatotic hepatocytes model.Up-regulated miR-22 expression negatively regulates the expression of SIRT1 to promote fat deposition in the human steatotic hepatocyte model.

关 键 词：非酒精性脂肪性肝病 miR-22 脂肪变 SIRT1 

分 类 号：R575.5[医药卫生—消化系统]