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:检测乙型肝炎病毒(hepatitis B virus,HBV)抗原和HBV DNA,比较壮药汗衣台的复方制剂与单味药在抗乙型肝炎病毒方面的疗效.方法:根据壮族民间常用的经验组方,不同浓度汗衣台及其配伍药分别与体外培养的2215细胞共同孵育,以XTT方法...目的:检测乙型肝炎病毒(hepatitis B virus,HBV)抗原和HBV DNA,比较壮药汗衣台的复方制剂与单味药在抗乙型肝炎病毒方面的疗效.方法:根据壮族民间常用的经验组方,不同浓度汗衣台及其配伍药分别与体外培养的2215细胞共同孵育,以XTT方法检测他们对2215细胞的毒性作用,选择各种药物对细胞无明显毒害的浓度进行组配;3d更换一次含药培养液,9d后收集上清液;用ELISA法测定上清液中乙型肝炎表面抗原(hepatitis B surface antigen,HBsAg)和乙型肝炎e抗原(hepatitis B e antigen,HBeAg)含量,荧光定量PCR法检测上清液中HBV-DNA的分泌.结果:复方组对HBsAg抗原的抑制作用稍高于单药组,但统计学上无显著意义;中低浓度的复方制剂F3-F5抑制HBeAg以及HBV-DNA分泌的作用明显强于相对应浓度的汗衣台单药组(P=0.002-0.009;P=0.038-0.05),而且抑制HBV-DNA分泌的疗效明显优于高浓度的复方制剂F1-F2(P=0.048-0.015).结论:组方配伍后,中低浓度的复方制剂抗病毒的综合疗效优于相对应浓度汗衣台单味药及高浓度复方制剂,各种药物成分相互影响、协同作用,不仅可以降低单个药物浓度,而且可以产生叠加增效作用.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.
文摘目的:检测乙型肝炎病毒(hepatitis B virus,HBV)抗原和HBV DNA,比较壮药汗衣台的复方制剂与单味药在抗乙型肝炎病毒方面的疗效.方法:根据壮族民间常用的经验组方,不同浓度汗衣台及其配伍药分别与体外培养的2215细胞共同孵育,以XTT方法检测他们对2215细胞的毒性作用,选择各种药物对细胞无明显毒害的浓度进行组配;3d更换一次含药培养液,9d后收集上清液;用ELISA法测定上清液中乙型肝炎表面抗原(hepatitis B surface antigen,HBsAg)和乙型肝炎e抗原(hepatitis B e antigen,HBeAg)含量,荧光定量PCR法检测上清液中HBV-DNA的分泌.结果:复方组对HBsAg抗原的抑制作用稍高于单药组,但统计学上无显著意义;中低浓度的复方制剂F3-F5抑制HBeAg以及HBV-DNA分泌的作用明显强于相对应浓度的汗衣台单药组(P=0.002-0.009;P=0.038-0.05),而且抑制HBV-DNA分泌的疗效明显优于高浓度的复方制剂F1-F2(P=0.048-0.015).结论:组方配伍后,中低浓度的复方制剂抗病毒的综合疗效优于相对应浓度汗衣台单味药及高浓度复方制剂,各种药物成分相互影响、协同作用,不仅可以降低单个药物浓度,而且可以产生叠加增效作用.
