丙型肝炎病毒F蛋白上调肝星形细胞胶原表达

Up-Regulation of Collagen Expression in Hepatic Stellate Cells by F Protein of Hepatitis C Virus

目的探讨丙型肝炎病毒(HCV)的F蛋白对肝星形细胞生长及胶原蛋白表达的影响。方法将含有HCV f基因的pcDNA3.1-f质粒或pcDNA3.1空载体分别转染肝星形细胞株LX2,G418筛选后获得稳定转染的细胞,分别命名为LX-f细胞或LX-p细胞,MTT法测定细胞增殖情况,ELISA法检测细胞分泌的Ⅰ型和Ⅲ型胶原蛋白含量。结果自培养24 h起,LX-f细胞各时间点的增殖率均大于LX-p细胞(P<0.01)。培养48 h时,LX-f细胞的Ⅰ型胶原蛋白及Ⅲ型胶原蛋白分泌量分别为(25.89±0.42)ng/ml和(18.21±0.49)ng/ml,明显高于LX-p细胞的(22.65±0.49)ng/ml和(15.29±0.62)ng/ml(P<0.01)。结论 HCV的F蛋白能促进肝星形细胞增殖,并上调细胞分泌Ⅰ型和Ⅲ型胶原蛋白,诱发肝纤维化。

Objective To investigate the effect of hepatitis C virus（HCV） F protein on proliferation and collagen expression of hepatic stellate cells.Methods After pcDNA3.1-f plasmid containing HCV f gene or empty pcDNA3.1 plasmid was transfected hepatic stellate cells LX2 by liposome,LX-f or LX-p cells were obtained by G418 screening.The proliferation of LX-f or LX-p cells was analyzed by MTT,and the contents of collagen type Ⅰand Ⅲ secreted by LX-f or LX-p cells were detected by ELISA.Results After 24 h cultivation,the proliferation rate of LX-f cells was higher than that of LX-p cells at each time point（P0.01）.After 48 h cultivation,the contents of collagen typeⅠand Ⅲ secreted by LX-f were（25.89±0.42） ng/ml and（18.21±0.49） ng/ml,which was significantly higher than those of LX-p cells（（22.65±0.49） ng/ml and（15.29±0.62） ng/ml）,P0.01.Conclusion HCV F protein is able to promote proliferation of hepatic stellate cells,and up-regulate the excretion of collagen type Ⅰand Ⅲ in those cells,which induces hepatic fibrosis.