目的探讨靶向肝细胞的反义RNA抗HBV作用。方法以糖化多聚赖氨酸为载体，将能转录HBV C区反义RNA的真核质粒pREP4-aC定向导入2.2.15细胞，潮霉素筛选阳性克隆。研究过程中用ELISA和斑点杂交方法检测多个时期培养上清液中HBsAg、HBeAg 和HBV DNA，同时观察对细胞的毒性作用。结果导向后24h出现对HBsAg、HBeAg和HBV DNA的抑制，6d左右抑制率达最高水平。未发现对2.2.15细胞的毒性作用。结论糖化多聚赖氨酸导向肝细胞的C区反义RNA，能有效抑制HBV抗原表达和DNA复制。%Objective To investigate the anti-HBV effect of targeted antisense RNA to hepatic cells. Methods pREP4-aC which would transcript antisense RNA against HBV C gene in eukaryotic cells were delivered into 2.2.15 cells by glactosylated poly-L-lisine(Gal-PLL), and the positive cells were selected. HBsAg, HBeAg and HBV DNA produced by 2.2.15 cells were detected with ELISA or Southern blot during the experiment, and the cytotoxicity of targeted antisense on 2.2.15 cells was observed. Results The inhibition effect on HBsAg, HBeAg and HBV DNA occurred at the 24th hour after delivery and reached the highest level at the 6th day, and kept at lest 2 months. No cytotoxicity on 2.2.15 cells was observed. Conclusion The targeted antisense against HBV C gene by delivery of Gal-PLL could effectively inhibit the antigen expression of HBV and DNA replication.
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