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

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

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

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

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/Abstract

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:

李玉香, 张凯宇, 张一宁, 王峰, 姜艳芳, 牛俊奇. 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[J]. J4, 2010, 36(3): 505-509.

LI Yu-Xiang, ZHANG Kai-Yu, ZHANG Yi-Ning, WANG Feng, JIANG Yan-Fang, NIU Dun-Qi. Effects of human hepatocyte growth factor gene on CCl4-injured human hepatocytes and rat hepatic stellate cell line and their mechanisms[J]. J4, 2010, 36(3): 505-509.

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