Objective To investigate the cleavage activities of ribozyme RCP in blocking HBV gene expression and replication in eukaryotic cells. Methods: The recombinant plasmid PCR/pSVL which can transiently express ribozyme RC...Objective To investigate the cleavage activities of ribozyme RCP in blocking HBV gene expression and replication in eukaryotic cells. Methods: The recombinant plasmid PCR/pSVL which can transiently express ribozyme RCP in eukaryotic cells was constructed and introduced into HepG2215 cell line with the technique of lipofectamine-mediated gene transfer. The vector pSVL transfection group served as the control. HB-sAg and HBsAg from culture medium of the cells were tested with solid-phase radioimmunoassay. Results:The cpm obtained from the medium samples showed that the inhibition rate of ribozyme RCP for HBsAg andHBeAg was 26. 7% and 24. 8% respectively in this transient expression system. Conclusion:Hammerhead ribozyme RCP can inhibit to some extent the expression of HBV gene.展开更多
目的:研究汗衣台体外抗乙型肝炎病毒(hepatitis B virus,HBV)的疗效.方法:不同浓度汗衣台与体外培养的2.2.15细胞共同孵育,以XTT方法检测其对2.2.15细胞的不良反应,以此决定汗衣台的安全浓度范围;3d更换一次含药培养液,9d后收集上清液;...目的:研究汗衣台体外抗乙型肝炎病毒(hepatitis B virus,HBV)的疗效.方法:不同浓度汗衣台与体外培养的2.2.15细胞共同孵育,以XTT方法检测其对2.2.15细胞的不良反应,以此决定汗衣台的安全浓度范围;3d更换一次含药培养液,9d后收集上清液;用ELISA法测定上清液中HBsAg和HBeAg含量,荧光定量PCR法检测上清液中HBV DNA的分泌.结果:各浓度汗衣台对HBsAg、HBeAg均有明显抑制作用,抑制率呈剂量依赖性:500-800μg/mL汗衣台对HBsAg的抑制明显高于同浓度的拉米夫定(P=0.002-0.000)及干扰素(P=0.006-0.003),100-800μg/mL浓度汗衣台对HBeAg的抑制明显高于同浓度拉米夫定(P=0.002-0.000)及干扰素(P=0.003-0.002),差异均有显著性.各浓度汗衣台对HBV DNA抑制亦呈剂量依赖性,500-800μg/mL浓度汗衣台短期内抑制HBV DNA分泌的疗效优于同浓度干扰素(P=0.018-0.031),但不如拉米夫定.结论:壮药汗衣台具有一定的体外抗HBV作用.展开更多
文摘Objective To investigate the cleavage activities of ribozyme RCP in blocking HBV gene expression and replication in eukaryotic cells. Methods: The recombinant plasmid PCR/pSVL which can transiently express ribozyme RCP in eukaryotic cells was constructed and introduced into HepG2215 cell line with the technique of lipofectamine-mediated gene transfer. The vector pSVL transfection group served as the control. HB-sAg and HBsAg from culture medium of the cells were tested with solid-phase radioimmunoassay. Results:The cpm obtained from the medium samples showed that the inhibition rate of ribozyme RCP for HBsAg andHBeAg was 26. 7% and 24. 8% respectively in this transient expression system. Conclusion:Hammerhead ribozyme RCP can inhibit to some extent the expression of HBV gene.
