反义X区RNA和反义P区RNA小鼠体内抗HBV的比较

目的:探讨针对乙型肝炎病毒(HBV)X区反义RNA和P区反义RNA对乙型肝炎转基因小鼠HBV复制和表达的影响.方法:构建表达HBV X区反义RNA和P区反义RNA的重组载体质粒,与脂质体混合,经尾静脉注入小鼠体内,用荧光聚合酶链反应定量法检测血清HBV DNA含量,用Nested-PCR法检测血清HBV DNA转阴率.结果:PLXSN-asX小鼠血清HBV DNA的复制于给药后2 d被抑制,至给药后8周抑制依然存在;PLXSN-asP小鼠血清HBV DNA的复制于给药后4 wk被抑制,至给药后8周抑制处于上升趋势. 与给药前相比,小鼠血清HBV DNA的复制,在PLXSN-asX组和PLXSN-asP组分别于2和8 wk达到抑制高峰,抑制率均为58%; 8周后小鼠血清HBV DNA转阴率PLXSN-asX组和PLXSN-asP组分别为25.0%(2/8), 12.5%(1/8).结论:HBV X区反义RNA与HBV P区反义RNA对乙型肝炎病毒转基因鼠抗HBV作用效率无显著差异,但X区反义RNA的起效时间明显早于P区反义RNA.

Comparison of antiviral effect between X-region and P-region antisense RNA in HBV transgenic mice

AIM: To investigate the effect of antisense RNA of hepatitis B virus (HBV) in X region and P region on replication and expression of HBV gene in HBV transgenic mice. METHODS: Recombinant vector plasmids expressing antisense RNA complementary to HBV X region and antisense RNA complementary to HBV P region were constructed and injected into transgenic mice through tail vein. The experimental mice were divided into blank, PLXSN, PLXSN-asX and PLXSN-asP groups. HBV DNA in serum was tested by fluorescent quantitative PCR and Nested-PCR. RESULTS: The expression of HBV DNA in serum was inhibited in PLXSN-asX group at day 2 after injection and still inhibited at week 8, while it was inhibited in PLXSN-asP group at week 4 and reached the lowest at week 8. In PLXSN-asX group and PLXSN-asP group, the strongest inhibitory effect on the expression of HBV DNA reached 58% at the 2nd week and 8th week after treatment respectively. The serum HBV DNA of 2/8 mice in the PLXSN-asX group and 1/8 mice in the PLXSN-asP group became negative.CONCLUSION: The antiviral effect of antisense RNA in X region of HBV was earlier than that of antisense RNA in P region of HBV. But the difference of antiviral effect between the 2 regions was not significant.