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

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。

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

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.