双靶区反义RNA抗乙型肝炎病毒的实验研究

目的 研究双靶区反义RNA重组载体质粒的转染、表达及抗乙型肝炎病毒的作用。方法 构建表达乙型肝炎病毒X、P双靶区正、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乙型肝炎病毒DNA含量，用Nested—PCR法检测血清乙型肝炎病毒DNA转阴率。结果 与给药前相比，小鼠血清乙型肝炎病毒DNA的复制，在逆转录病毒载体-asX组和逆转录病毒载体-asP组分别于第2、第8周达到抑制高峰，抑制率均为58％；在逆转录病毒载体-asXP组于第1、第8周出现两次抑制高峰，抑制率分别为66％、77%；其余组未出现明显变化。给药后8周，小鼠血清乙型肝炎病毒DNA，在逆转录病毒载体-asX组、逆转录病毒载体-asP和逆转录病毒载体-asXP分别有2只、1只，1只转阴，转阴率分别为25.0％、12．5％、16．7％。结论 乙型肝炎病毒X、P双靶区反义RNA对乙型肝炎病毒转基因鼠乙型肝炎病毒DNA的抑制作用优于单靶区反义RNA，但其对血清乙型肝炎病毒DNA的转阴率与单靶区反义RNA无显著性差异。

Experimental study on anti-hepatitis B virus by dual-target antisense RNA in HBV transgenic mice

Objective To study the effects of dual-target antisense RNA in HBV transgenic mice.Methods Recombinant vector plasmids expressing dual-target antisense RNA complementary to HBV X and P region were constructed and injected into transgenic mice through tail vein.The experimental mice were divided into blank,PLXSN,PLXSN-asX,PLXSN-asP,PLXSN-asXP and PLXSN-seXP groups.HBV DNA in serum was tested by FQ-PCR and Nested-PCR.Results The expression of HBV DNA in serum was inhibited distinctively(58%) in PLXSN-asX group in the 2nd week while in PLXSN-asP group in the 8th week,and in PLXSN-asXP group the inhibitory effect on the expression of HBV DNA reached 66% in the 1st week and 77% in the 8th week after treatment. In the 8th week after treatment,the expression of HBV DNA in serum became negative in 2/8 of PLXSN-asX group,in 1/8 of PLXSN-asP group and in 1/6 of PLXSN-asXP group,while the expression of HBV DNA in serum was all positive in other groups.Conclusion The inhibitory effect of dual-target antisense-RNA is stronger than that of single-target antisense-RNA,but no significant was found in negative transfer rate between dual-target antisense-RNA and single-target antisense-RNA.