J4 ›› 2010, Vol. 36 ›› Issue (3): 505-509.

• 基础研究 • 上一篇    下一篇

人肝细胞生长因子基因表达对CCl4损伤的人肝细胞及大鼠肝星状细胞株的影响及其机制

李玉香, 张凯宇, 张一宁, 王峰, 姜艳芳, 牛俊奇   

  1. 吉林大学第一医院感染症科, |吉林 长春 130021
  • 收稿日期:2009-12-30 出版日期:2010-05-28 发布日期:2010-05-28
  • 通讯作者: 牛俊奇 (Tel:0431-88782413,E-mail: junqiniu@yahoo.com.cn) E-mail:junqiniu@yahoo.com.cn
  • 作者简介:李玉香(1964-)|女|吉林省长春市人|副教授|医学博士|主要从事感染性疾病及病毒性肝炎的研究。
  • 基金资助:

    吉林省科技厅科研基金资助课题(20060417-2)

Effects of human hepatocyte growth factor gene on CCl4-injured human hepatocytes and rat hepatic stellate cell line and their mechanisms

LI Yu-Xiang, ZHANG Kai-Yu, ZHANG Yi-Ning, WANG Feng, JIANG Yan-Fang, NIU Dun-Qi   

  1. Department of  |Infectious Diseases,First Hospital,Jilin University,Changchun 130021,China
  • Received:2009-12-30 Online:2010-05-28 Published:2010-05-28

摘要:

目的:探讨人肝细胞生长因子(hHGF) 基因的表达对CCl4损伤的人肝细胞及大鼠肝星状细胞株(CFSC-2G)生物学效应的影响及其对CCl4损伤的人正常肝细胞株(HL-7702)的保护作用,进一步探讨凋亡产生的机制。方法:将获得的稳定转染HL-7702细胞克隆,分为4组,Ⅰ组为转染PCI-neo-hHGF实验组,Ⅱ组为转染PCI-neo空载体对照组,Ⅲ组为仅加入脂质体的对照组,Ⅳ组为空白对照组。采用MTT法测定细胞增殖活性。采用Western blotting测定转染PCI-neo-hHGF的HL-7702细胞和CFSC-2G细胞中hHGF、caspase-3、caspase-8、caspase-9蛋白质的表达。结果:HL-7702细胞转染PCI-neo-HGF并培养48 h后,细胞的增殖活性明显高于PCI-neo空载体对照组、脂质体转染对照组和空白对照组HL-7702细胞(P<0.01),而且随着转染PCI-neo-HGF剂量增加,细胞的增殖活性有一定的增长趋势,但各组间比较差异无显著性(P>0.05);PCI-neo-HGF转染的HL-7702细胞caspase-3蛋白质表达量较CCl4损伤的HL-7702细胞和PCI-neo转染的HL-7702细胞明显降低,未检测到caspase-8和caspase-9蛋白质的表达;PCI-neo-HGF转染的CFSC-2G细胞caspase-3蛋白质表达量较CCl4诱导的CFSC-2G增加,caspase-9蛋白质表达也呈阳性。结论:PCI-neo-HGF可促进体外培养的HL-7702细胞的生长增殖,能够抑制CCl4损伤的HL-7702细胞的凋亡,促进大鼠CFSC-2G细胞的凋亡,其作用机制可能与上调caspase-3和caspase-9蛋白质表达有关。

关键词: 人肝细胞生长因子; CFSC-2G细胞;细胞凋亡;caspase-3;caspase-8;caspase-9

Abstract:

Abstract:Objective To explore the biological effects of human hepatocyte growth factor (hHGF) gene expression on CCl4-injured human hepatocytes and rat hepatic stellate cell line and the protective effect of hHGF gene expression on CCl4-injured human normal liver cell line (HL-7702),and   explore the mechanism of apoptosis.Methods The  stably transfected HL-7702 cell clones were divided into four groups:PCI-neo-hHGF  group,   PCI-neo empty vector group,liposome control group,and blank control group.The proliferative activity of cells was determined by MTT method.The protein expressions of hHGF,caspase-3,caspase-8, and caspase-9 were measured using Western blotting in PCI-neo-hHGF transfected HL-7702 cells and CFSC-2G cells.Results After the HL-7702 cells were transfected with PCI-neo-HGF and cultivated for 48 h,the cell proliferation activity was significantly higher than those in  PCI-neo,liposome and control groups (P<0.01);and with the increasing of transfected dose of PCI-neo-HGF,the proliferation activity of the cells had a trend of increasing,but there was no significant difference between various groups(P>0.05).The protein expression of caspase-3 in PCI-neo-HGF transfected HL-7702 cells was decreased significantly compared with those in CCl4-injured HL-7702 cells and PCI-neo transfected HL-7702 cells;the protein expressions of caspase-8 and caspase-9 were not detected;the protein expression of caspase-3 in PCI-neo-HGF transfected CFSC-2G cells was increased compared with those in CCl4-induced CFSC-2G cells,the protein expression of caspase-9 was also positive.Conclusion PCI-neo-HGF can promote the growth and proliferation of HL-7702 cells,and inhibit  the  apoptosis of CCl4-injured HL-7702 cells,and promote the apoptosis of  rat CFSC-2G cells  in vitro.Its mechanism may be related to the up-regulation of the protein expressions of caspase-3 and caspase -9.

Key words: human hepatocyte growth factor;CFSC-2G cells;apoptosis;caspase-3;caspase-8;caspase-9

中图分类号: 

  • Q78