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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 effects of hepatitis C virus (HCV) F protein on hepatic stellate cell growth and collagen expression. Methods The pcDNA3.1-f plasmid or pcDNA3.1 empty vector containing HCV f gene was transfected into LX-2 cells, respectively, and then transfected into LX-f cells or LX-p cells Cell proliferation was measured by MTT assay. Collagen type I and III secreted by cells were detected by ELISA. Results The proliferation rate of LX-f cells was higher than that of LX-p cells at 24 h after culture (P <0.01). After cultured for 48 h, the levels of collagen Ⅰ and collagen Ⅲ in LX-f cells were (25.89 ± 0.42) ng / ml and (18.21 ± 0.49) ng / ml, respectively, which were significantly higher than those in LX-p cells 22.65 ± 0.49 ng / ml and 15.29 ± 0.62 ng / ml respectively (P <0.01). Conclusion The F protein of HCV can promote the proliferation of hepatic stellate cells and upregulate the secretion of collagen type Ⅰ and Ⅲ and induce hepatic fibrosis.