摘要
研究以宿主基因ASGPR为靶的反义核酸与抗乙肝病毒(hepatitis B virus,HBV)药物联合应用的抗-HBV作用,为HBV感染的联合治疗和用药提供新的思路。以HepG2.2.15细胞为靶细胞,脂质体为载体将靶向ASGPR的硫代反义寡核苷酸(ASODN)转染至HepG2.2.15细胞中,6h后加入抗病毒药拉米夫定(3TC)或阿地福韦(ADV),72h后收集细胞培养液,采用酶联免疫法及荧光定量PCR法测定ASODN与抗-HBV药物联合应用后对细胞培养液中HBsAg、HBeAg及细胞分泌HBV DNA的抑制作用。采用金正均Q值法对数据进行分析。拉米夫定(3TC)和阿地福韦(ADV)分别与ASODN联用对HBsAg的抑制呈相加或协同作用。随着ADV浓度的增加,两药的协同作用下降。3TC与ASODN联用对HBeAg的抑制呈拮抗、相加和协同作用,其拮抗作用随着3TC浓度的增加逐渐减弱。ADV与ASODN联用对HBeAg的抑制只呈相加作用。3TC和ADV分别与ASODN联用对HBV DNA的抑制均表现为相加作用或协同作用,取ADV0.5μmol/L与取ASODN0.2μmol/L联用时对HBV DNA的抑制率可提高到72.6%。ASODN在一定条件下分别与3TC及ADV联用有相加或协同抗HBsAg、HBeAg及HBV DNA的表达,尤以抗HBsAg作用显著增强。
This paper was to investigate the inhibitory effects against HBV of a phosphorothioate antisense oligodeoxynucleotide ＂ASODN＂ targeted to ASGPR combined with anti-HBV drugs in vitro. The assay attempted to access a new method and idea to refresh the thinking way about hepatitis B treatment by the combination of ASODN and drugs. A phosphorothioate antisense oligodeoxynucleotide targeted to asialoglycoprotein receptor （ASGPR） mRNA was synthesized. Using HepG2.2.15 cell as the target cell, ASODN was transfected into HepG2.2.15 cell by lipofectin. Six hours after the incubation of ASODNs, the medium was replaced with different concentrations of Lamivudine（3TC） and Adefovir Dipivoxil（ADV） and cultures were incubated for 72h. Then cell media were collected and detected. HBV DNA level in cell medium was examined by real-time PCR. HBsAg and HBeAg were detected by ELISA. Jin Zhengjun Method was applied to evaluate the interaction of ASODN and the anti-HBV drugs. Results showed that 3TC or ADV accompanying with ASODN separately showed an additive or synergistic effect towards the inhibition of HBsAg. With the increasing of ADV concentration, the synergistic effect between the two drugs declined gradually. The combination of 3TC and ASDON showed antagonistic, additive or synergistic effects on HBeAg. Especially, antagonistic effect decreased gradually along with the increasing of 3TC concentration. Meanwhile, the combined use of ADV and ASODN demonstrated the additive effect only on the inhibitive effect of HBeAg. The combined process of 3TC and ADV respectively with ASDON displayed the additive or synergistic effect on suppressing of HBV DNA. When ADV 0.5μmol/L combined with ASODN 0.2μmol/L, the inhibitive rate against HBV DNA could be promoted to 72.6 %. It is concluded that under certain experimental conditions, the combination of ASODN with 3TC and ADV respectively displayed the additive or synergistic effect against HBV. Especially the inhibition of HBsAg was significant.
出处
《病毒学报》
CAS
CSCD
北大核心
2006年第1期17-21,共5页
Chinese Journal of Virology
基金
军队"十五"重点项目(01z019)
国家863资助项目(2002AA2310512004AA234031)
国家自然科学基金项目(30171111)
国家重点基础研究发展计划(973)(2004CB518904)
关键词
寡核苷酸
反义
乙型肝炎病毒
联合治疗
协同作用
antisense oligodeoxynucleotides
hepatitis B virus
combined therapy
synergistic effect
