用重组8型腺相关病毒载体介导的乙型肝炎病毒持续感染小鼠模型评价核苷类似物的抗病毒效果

Anti-HBV effect of nucleotide analogues on mouse model of chronic HBV infection mediated by recombinant adeno-associated virus 8

摘要:

探索用重组8型腺相关病毒载体携带1.3拷贝乙型肝炎病毒 (rAAV8-1.3HBV) 介导的HBV持续感染小鼠模型评价核苷酸类似物抗病毒药物的抗病毒效果。首先，通过将rAAV8-1.3HBV经尾静脉注射到30只C57BL/6小鼠体内，建立HBV持续感染模型并对模型成功率进行检测，将建模成功的27只小鼠随机分成6组。然后采取灌胃的方式给予不同剂量的抗病毒药物恩替卡韦 (ETV) 及拉米夫定 (LAM)，每日1次，连续10 d，后停药15 d，同时设置生理盐水及空白对照组。其中ETV分为高剂量 (1.0 mg/(kg?d)) 和低剂量 (0.1 mg/(kg?d)) 两组；LAM分为高剂量 (500 mg/(kg?d)) 和低剂量 (100 mg/(kg?d)) 两组。检测给药前后和停药前后小鼠模型血清中HBV DNA、HBeAg和HBsAg表达水平并比较变化情况。结果发现连续给药10 d后，各给药组与生理盐水组相比，血清中HBV DNA水平均显著下降，具有统计学差异 (P＜0.05)。停药15 d后，低剂量的ETV与LAM两组血清HBV DNA水平出现反弹，差异存在统计学意义 (P＜0.05)。在整个实验过程中，各组小鼠血清中HBeAg和HBsAg表达水平均未出现明显变化。上述结果表明，ETV和LAM能有效抑制模型小鼠中HBV病毒的复制，而对HBeAg和HBsAg表达水平无明显影响；提示AAV8-1.3HBV介导的HBV持续感染小鼠模型制备简单，成模率高，可有效体现出ETV和LAM抗HBV的作用效果，从而用于核苷酸类似物抗HBV药物的筛查。

Abstract:

We evaluated the anti-HBV effects of nucleotide analogues, Entecavir (ETV) and Lamivudine (LAM) targeting mouse model of HBV persistent infection with recombinant adeno-associated virus 8 carrying 1.3 copies of HBV genome (rAAV8-1.3HBV). Ninety percent (27 of 30 mice) of rAAV8-treated mice were chosen as mouse model. Four groups were orally administrated with different doses of ETV (1 mg/(kg?d) or 0.1 mg/(kg?d)) and LAM (500 mg/(kg?d) or 100 mg/(kg?d)) once a day for 10 days. The other two groups were set as normal saline treated and untreated control. We detected the levels of HBV DNA, HBeAg and HBsAg in sera at different time. Results indicate that HBV DNA level decreased significantly (P<0.05) in drug-treated groups compared with normal saline group after drug administration. Fifteen days after the drug withdrawal, HBV DNA level rebounded back obviously (P<0.05) in groups with low doses of ETV and LAM. However, there was no apparent change of HBeAg and HBsAg in the whole process among all groups. These results showed that our model could reflect the anti-viral effect of nucleotide analogues. This model can be a useful and convenient tool for anti-HBV drug discovery.