摘要目的 研究HepG2.2.15细胞株在体外能否促进肝星状细胞(HSC)中肝纤维化相关因子的表达,进而探索HBV促肝细胞纤维化的机制.方法 将HepG2和HepG2.2.15细胞株分别在体外与HSC共培养,以单独培养的HSC为对照组.取培养后24、48和72 h 3个时间点,以实时定量PCR检测HSC中基质金属蛋白酶-2(MMP-2)和金属蛋白酶组织抑制剂-1(TIMP-1)mRNA的表达;以免疫印迹法定量检测HSC中MMP-2和TIMP-1的表达.结果 与对照组和与HepG2共培养的HSC比较,与HepG2.2.15细胞株共培养的HSC中MMP-2和TIMP-1 mRNA的表达明显增高,以72 h的差异最为显著(F值分别为11.91和23.13,P值分别为0.008和0.001);与HepG2.2.15共培养的HSC中MMP-2和TIMP-1蛋白的表达亦明显增高(F值分别为20.70和6.54,P值分别为0.002和0.03).结论 与HepG2.2.15细胞株共培养后,HSC中肝纤维化相关因子的表达明显增强.体外试验证明HBV具有诱导肝细胞纤维化的重要作用.

abstractsObjective To investigate the effect of HBV on the expression of fibrosis-related factors in hepatic stellate cells(HSC)and its relation with liver fibrosis.Methods HSCs were co-cultured with HepG2 or HepG2.2.15 in vitro and HSCs cultured alone served as the control.The mRNA expression of matrix metalloproteinase(MMP)-2 and tissue inhibitor of metalloproteinase(TIMP)-1 was detected by realtime PCR.The protein expression of MMP-2 and TIMp-1 was detected by Western-blot.Results Compared with the control and the HSCs co-cultured with HepG2,the expression of MMP-2 and TIMP-1 mRNA in HSCs co-cultured with HepG2.2.15 was increased remarkably and the most significant difference was found at 72 h(F=11.91,23.13;P=0.008,0.001);the expression of MMP-2 and TIMP-1 protein in HSCs co-cuhured with HepG2.2.15 was also increased remarkably and the most significant difference was found at 72 h(F=20.70,6.54;P=0.002,0.003)too.Conclusion The expression of fibrosis-related factors in HSCs increased significantly after co-cultured with HepG2.2.15,which suggests that HBV could promote liver fibrosis.