体外诱导的酒精性脂肪肝细胞模型的建立、鉴定及机制探讨  

Establishment and identification of alcohol induced steatotic hepatocytes model

作　　者：廖于[1] 李龙辉[2] 左国庆[2] 吴海峰[1] 秦观海[1] 宋世卿[1] 袁英[1] 邱烈[1] 周天寒[1] 朱照静[1] 

机构地区：[1]重庆医药高等专科学校基础部,400051 [2]重庆医科大学附属第二医院消化内科,400010

出　　处：《重庆医学》2010年第8期902-904,I0001,共4页Chongqing medicine

基　　金：重庆市卫生局医学科研计划资助项目(2009-2-445)

摘　　要：目的建立体外诱导的酒精性脂肪肝细胞模型,初步探讨酒精性脂肪肝发生的机制。方法采用MTT法观察乙醇对人正常肝细胞株L02生长的影响,筛选最佳乙醇诱导浓度,全自动生化仪检测细胞内三酰甘油(TG)水平,油红O染色法观察细胞内脂滴,透射电镜观察细胞超微结构,免疫细胞化学方法观察PPARγ、SREBP-1、SCAP的表达。结果MTT法筛选出浓度0.28%的乙醇作为建立体外诱导的酒精性脂肪肝细胞模型的作用浓度;与L02细胞相比,0.28%乙醇诱导至第30代细胞(Ld30细胞)内TG显著增多(P<0.05);油红O染色发现Ld30细胞内大量桔红色颗粒聚积;透射电镜观察超微结构发现胞浆中有许多大小不一的脂滴。Ld30细胞PPARγ、SREBP-1、SCAP的MOD值和阳性表达指数均显著增高(P<0.05)。结论本研究在体外成功建立酒精性脂肪肝的细胞模型,提示酒精性脂肪肝发生的机制可能与PPARγ、SREBP-1、SCAP上调有关。Objective To establish and identify the alcohol induced steatotic hepatocytes model,and to explore the mechanism. Methods The effect of alcohol on a normal hepatocytes line, L02,was observed using MTT method. The best concentration of alcohol was screened out. The triglyceride （TG） concentration was measured by Automatic Biochemical Tester and Analyzer,the lipid droplets were observed by Oil Red O Method, the ultrastructure of the hepatocytes were observed by Transmission Electron Microscope,and the expression of PPART,SREBP-1 and SCAP was observed by immunocytochemical method. Results The 0.28% alcohol was chosen as the best concentration of alcohol for the induction of steatotic hepatocytes. Comparing with normal 1.02 cells,TG was significantly increased in the Alcohol induced steatotic hepatocytes of passage 30 （Ld30） （P〈0.05）. Lots of orange particles, which were stained by Oil Red O,were found inside the Ld30 cells. Many lipid droplets with different size were found inside the cytoplasm when observed by TEM. And the MOD value and index of positive cells of the PPART.SREBP 1 .SCAP for the Ld30 cells were significantly increased （P〈0.05）. Conclusion An alcohol induced steatotic hepatocytes model is established successfully. The possible mechanism for the hepatocytes steatosis can be related with the upregulation of PPART,SREBP-1 and SCAP.

关 键 词：酒精性脂肪肝 细胞模型 机制 

分 类 号：R575.5[医药卫生—消化系统]