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浙江大学学报(医学版)  2016, Vol. 45 Issue (1): 68-74    DOI: 10.3785/j.issn.1008-9292.2016.01.11
原著     
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
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3, 刘恩岐1,2
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
Overexpression of PPARγ induces adipogenic steatosis in mouse primary hepatocytes
BAI Liang1,2, ZHANG Yali1,2, XIE Chen1,2, WANG Rong1,2, ZHAO Sihai1,2, JIA Yuzhi3, LIU Enqi1,2
1. Research Institute of Atherosclerotic Disease, Xi'an Jiaotong University School of Medicine, Xi'an 710061, China;
2. Laboratory Animal Center, Xi'an Jiaotong University School of Medicine, Xi'an 710061, China;
3. Department of Pathology, Feinberg School of Medicine, Northwestern University, Chicago, IL 60611, USA
全文: PDF(990 KB)  
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。

关键词 小鼠,近交C57BL肝细胞/细胞学PPARγ/生物合成脂肪生成细胞,培养的    
Abstract

Objective: To investigate the effects of PPARγ overexpression on steatosis in mouse primary hepatocytes. Methods: Primary hepatocytes isolated from C57BL/6J mice were infected with either Ad/LacZ or Ad/PPARγ for 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 PPARγ and its target aP2 strongly increased. Conclusion: over expression of PPARγ induces adipogenic steatosis in mouse primary hepatocytes.

Key wordsMice, inbred C57BL    Hepatocytes/cytology    PPAR gamma/biosynthesis    Lipogenesis    Cells, cultured
收稿日期: 2015-09-30     
CLC:  R575  
基金资助:

国家自然科学基金(81200207);中央高校基本科研业务费专项资金

通讯作者: 刘恩岐(1965-),男,博士,教授,从事动物模型与动脉粥样硬化研究;E-mail:liuenqi@mail.xjtu.edu.cn;http://orcid.org/0000-0002-2092-3158     E-mail: liuenqi@mail.xjtu.edu.cn
作者简介: 白亮(1981-),女,博士,讲师,从事脂质代谢与动脉粥样硬化研究;E-mail:bailiang0922@mail.xjtu.edu.cn;http://orcid.org/0000-0001-7972-0892
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白亮 等. 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[J]. 浙江大学学报(医学版), 2016, 45(1): 68-74.
BAI Liang, ZHANG Yali, XIE Chen, WANG Rong, ZHAO Sihai, JIA Yuzhi, LIU Enqi. Overexpression of PPARγ induces adipogenic steatosis in mouse primary hepatocytes. Journal of ZheJiang University(Medical Science), 2016, 45(1): 68-74.

链接本文:

http://www.zjujournals.com/xueshu/med/CN/10.3785/j.issn.1008-9292.2016.01.11      或      http://www.zjujournals.com/xueshu/med/CN/Y2016/V45/I1/68

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