上海交通大学学报（医学版）

上海交通大学学报（医学版）

• 论著（基础研究） • 上一篇    下一篇

分子伴侣4-苯基丁酸对HepG2脂肪变模型中细胞损伤的影响

范竹萍，艾罗燕，陈荔萍，吴昌维，陈志威，苏大芝，许青青，王晓晗   

  1. 上海交通大学 医学院附属仁济医院健康保健中心 上海市消化疾病研究所， 上海 200127
  • 出版日期:2014-12-28 发布日期:2014-12-30
  • 作者简介:范竹萍（1965—）， 女， 主任医师， 博士， 硕士生导师; 电子信箱: zhuping_fan@163.com。
  • 基金资助:

    上海市公共卫生重点学科项目（12GWZX0903）

Effects of molecular chaperone 4-phenylbutyric acid on cell damage of steatotic HepG2 cells

FAN Zhu-ping, AI Luo-yan, CHEN Li-ping, WU Chang-wei, CHEN Zhi-wei, SU Da-zhi, XU Qing-qing, WANG Xiao-han   

  1. Department of Health Care Center, Renji Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai Institute of Digestive Disease, Shanghai 200127, China
  • Online:2014-12-28 Published:2014-12-30
  • Supported by:

    Key Discipline Construction Project of Shanghai Public Health，12GWZX0903

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摘要：

目的 观察人肝癌HepG2细胞脂肪变模型中内质网应激发生的情况，探讨分子伴侣4-苯基丁酸（4-PBA）对该脂肪变模型中内质网应激、氧化应激和凋亡的影响。方法 使用0.5、1.0 mmol/L的混合脂肪酸分别干预HepG2细胞，在不同时间点采用Real-time PCR方法检测内质网应激相关蛋白CHOP和葡萄糖调节蛋白78 （GRP78）mRNA的表达水平。以2 mmol/L的4-PBA干预脂肪变性HepG2细胞，在不同时间点检测CHOP和GRP78 mRNA的表达，氧化应激指标丙二醛(MDA)含量及超氧化物歧化酶（SOD）和还原型谷胱甘肽（GSH）水平，以及凋亡相关蛋白Caspase-3活性的变化。结果 与对照组比较，混合脂肪酸干预组HepG2细胞内CHOP、GRP78 mRNA的表达水平显著升高（P<0.05）。4-PBA干预可使脂肪变性HepG2细胞内CHOP和GRP78 mRNA的表达水平下调（P<0.05），MDA含量明显下降，SOD和GSH水平显著升高（P<0.05），Caspase-3活性减低（P<0.05）。结论 HepG2细胞脂肪变性时发生明显的内质网应激。分子伴侣4-PBA能减轻氧化应激和内质网应激，下调脂肪变性HepG2细胞Caspase-3活性，减轻细胞损伤。

关键词: 脂肪变性HepG2细胞, 氧化应激, 内质网应激, Caspase-3, 4-苯基丁酸

Abstract:

Objective To observe the occurrence of endoplasmic reticulum stress in steatotic HepG2 cells and to explore the effects of chemical molecular chaperone 4-phenylbutyric acid (4-PBA) on the endoplasmic reticulum stress, oxidative stress, and apoptosis of steatotic HepG2 cells. Methods HepG2 cells were intervened by fatty acid mixtures of 0.5 mmol/L and 1.0 mmol/L. The mRNA expressions of endoplasmic reticulum stress related protein CHOP and glucose regulated protein 78 (GRP78) were detected by the Real-time PCR at different time points. Then steatotic HepG2 cells were intervened by 4-PBA of 2 mmol/L. At different time points, the mRNA expressions of CHOP and GRP78 and levels of malonaldehyde (MDA), which was the oxidative stress index, superoxide dismutase (SOD), and reduced glutathione (GSH) were detected and variations of the viability of apoptosis related protein Caspase-3 were observed. Results Compared to the control group, the mRNA expressions of CHOP and GRP78 in HepG2 cells of fatty acid intervened group increased significantly (P<0.05). The intervention of 4-PBA decreased the mRNA expressions of CHOP and GRP78 in steatotic HepG2 cells (P<0.05) and MDA level; significantly increased SOD and GSH levels (P<0.05); and decreased the viability of Caspase-3 (P<0.05). Conclusion Significant endoplasmic reticulum stress occurs in steatotic HepG2 cells. Molecular chaperone 4-PBA may alleviate oxidative stress and endoplasmic reticulum stress, decrease the viability of Caspase-3 of steatotic HepG2 cells, and alleviate the cell damage.

Key words: steatotic HepG2 cells, oxidative stress, endoplasmic reticulum stress, Caspase-3, 4-phenylbutyric acid