高表达过氧化物酶体增殖子激活受体γ诱导小鼠原代肝细胞发生脂肪变性  

Overexpression of PPARγinduces adipogenic steatosis in mouse primary hepatocytes

作　　者：白亮[1,2] 张亚莉[1,2] 谢晨[1,2] 王蓉[1,2] 赵四海[1,2] 贾玉枝 刘恩岐[1,2] 

机构地区：[1]西安交通大学医学部心血管中心动脉粥样硬化与脂代谢研究室,陕西西安710061 [2]西安交通大学医学部医学实验动物中心,陕西西安710061 [3]Department of Pathology, Feinberg School of Medicine, Northwestern University, Chicago, IL 60611, USA

出　　处：《浙江大学学报（医学版）》2016年第1期68-74,共7页Journal of Zhejiang University(Medical Sciences)

基　　金：国家自然科学基金(81200207)；中央高校基本科研业务费专项资金

摘　　要：目的：研究外源过氧化物酶体增殖子激活受体（ PPAR ）γ高表达对小鼠原代肝细胞脂肪变性的影响。方法：从5～6周龄C57BL／6J小鼠分离培养原代肝细胞，分别用LacZ腺病毒（ Ad／LacZ）或PPARγ腺病毒（ Ad／PPARγ）感染细胞48 h，油红O染色检测原代肝细胞脂肪积聚情况；实时定量PCR和蛋白质印迹法分析PPARγ、成脂相关基因aP2和CideA等mRNA和蛋白表达水平。结果：原代分离培养的小鼠肝细胞透光度好，胞核圆且透亮，细胞呈圆形生长，有双核，连接成片状或岛状。用Ad／LacZ或Ad／PPARγ感染48 h后，Ad／LacZ组肝细胞几乎无脂肪积聚，而Ad／PPARγ感染肝细胞中有大量脂肪滴沉积。外源PPARγ刺激作用下，肝细胞中PPARγ、aP2、FGF21、CideA mRNA表达增加，而adiponectin mRNA表达下降（均P＜0.05）。 Ad／PPARγ感染后，PPARγ和aP2蛋白表达也增加。结论：高表达PPARγ诱导小鼠原代肝细胞脂肪变性和脂肪相关基因表达。Objective： To investigate the effects of PPARγoverexpression on steatosis in mouse primary hepatocytes.Methods： Primary hepatocytes isolated from C57BL/6J mice were infected with either Ad/LacZ or Ad/PPARγfor 48 h.Steatosis of the primary hepatocytes was checked by Oil Red O staining.The mRNA and protein expression of adipocyte-specific genes PPARγ, aP2 and CideA were analyzed by using RT Real-time PCR and Western Blot.Results： Primary hepatocytes were small and even.Hepatocyte nuclei were round with dispersed chromatin and prominent nucleoli. Accumulated lipid droplets were observed in Ad/PPARγ-infected hepatocytes, but in Ad/LacZ-infected hepatocytes. Moreover, compared with Ad/LacZ-infected hepatocytes, the mRNA expression of PPARγ, aP2, FGF21 and CideA in Ad/PPARγ-infected hepatocytes were significantly induced, the protein expression of PPARγand its target aP2 strongly increased. Conclusion： over expression of PPARγinduces adipogenic steatosis in mouse primary hepatocytes.

关 键 词：肝细胞/细胞学 PPARγ/生物合成 脂肪生成 细胞 培养的 

分 类 号：R575[医药卫生—消化系统]