丙型肝炎病毒对SMMC 7721细胞体外直接致病作用的研究

Study of Hepatitis C Virus Direct Pathogenesis on SMMC 7721 in Vitro

目的:探讨丙型肝炎病毒(HCV)的直接致病作用。方法:用HCV阳性血清体外感染SMMC 7721细胞,观察细胞形态的变化,并运用四甲基偶氮唑盐(MTT)法及流式细胞仪(FCM)分别检测培养细胞的存活率及凋亡率,同时用套式逆转录-聚合酶链反应(nest RT-PCR)检测其中的正、负链HCV RNA。结果:HCV感染后部分SMMC7721细胞发生皱缩、脱壁,8h细胞存活率为74.2％,细胞凋亡率为18.7％,至56h后细胞存活率回复至84.3％以上,而细胞凋亡率又降至7.4％以下;各时间组的细胞存活率显著低于未感染HCV的SMMC7721细胞,而凋亡率则显著高于未感染的SMMC 7721细胞。感染后24h细胞及上清中HCV RAN检测结果均阴性,48h后在细胞中可检出正、负链HCV RNA,96h后细胞中HCV RNA正链呈阳性、负链转为阴性。结论:1.HCV具有一定的直接致病作用;2.HCV感染SMMC7721后的复制不稳定。

Objective, To investigate the direct pathogenesis of hepatitis C virus (HCV). Methods: After SMMC 7721 cells were infected by HCV positive serum, the change of cell's conformation was observed. RT-PCR, MTT and flow cytometry(FCM) were used to detect HCV RNA, the rate of cell livability and apotosis. Results: After HCV infected SMMC 7721, parts of cells wrinkled and fell off from the wall of the culture bottle. Although cell persistence was 74. 2% and the rate of cell apotosis was 18. 7% in 8h, the former was up to more than 84. 3% and the latter was down to less than 7. 4% after 56h. Contrasting with SMMC 7721 cells not infected by HCV, cell persistence was significantly lower, and the rate of cell apotosis was significantly higher in each time group. Plus and minus strand HCV RNA was negative at 24h, positive after 48h or 96h, while minus strand HCV RNA become negative after 96h. Conclusions HCV has definite direct pathogenesis. HCV replication in SMMC 7721 is not steady.