Xiao S R, Xu G D, Wei W J, Peng B, Deng Y B
Centre for Medical Laboratory Science, the Affiliated Hospital of Youjiang Medical College for Nationalities, Baise 533000, China.
Zhonghua Gan Zang Bing Za Zhi. 2018 Jan 20;26(1):17-22. doi: 10.3760/cma.j.issn.1007-3418.2018.01.006.
To investigate the antiviral effect of hepatitis B virus (HBV) S gene-specific anti-gene locked nucleic acid (LNA) in transgenic mice. A total of 30 HBV transgenic mice were randomly divided into blank control group (5% glucose + liposome), unrelated sequence control group, lamivudine control group, antisense LNA control group, and anti-gene LNA group, with 6 mice in each group. The mice in the lamivudine group were given lamivudine by gavage, and LNA was injected via the caudal vein. Quantitative real-time PCR was used to measure serum HBV DNA, ELISA was used to measure serum HBsAg, RT-PCR was used to measure HBV S mRNA level in the liver, and immunohistochemistry was used to measure the level of HBsAg in hepatocytes. At 3, 5, and 7 days after treatment, there were significant changes in the inhibition rates of HBV DNA (37.18%, 50.27%, and 61.46%, respectively) and HBsAg (30.17%, 44.00%, and 57.76%, respectively) achieved by anti-gene LNA ( < 0.01), and there were significant differences between the anti-gene LNA group and the other four control groups ( < 0.05). In the anti-gene LNA group, the relative mRNA expression of HBV S gene was 0.33 and the percentage of HBsAg-positive hepatocytes was 31%, which were significantly different from these two indices in the control groups ( < 0.05). There were no abnormal changes in liver/renal biochemical parameters and HE staining results. Anti-gene LNA targeting at HBV S gene has a strong antiviral effect in transgenic mice, which provides theoretical and experimental bases for gene therapy for HBV.
研究乙型肝炎病毒（HBV）S基因特异性抗基因锁核酸（LNA）对转基因小鼠的抗病毒作用。将30只HBV转基因小鼠随机分为空白对照组（5%葡萄糖+脂质体）、无关序列对照组、拉米夫定对照组、反义LNA对照组和抗基因LNA组，每组6只。拉米夫定组小鼠通过灌胃给予拉米夫定，LNA通过尾静脉注射。采用定量实时PCR检测血清HBV DNA，ELISA检测血清HBsAg，RT-PCR检测肝脏中HBV S mRNA水平，免疫组化检测肝细胞中HBsAg水平。治疗后3、5、7天，抗基因LNA对HBV DNA的抑制率（分别为37.18%、50.27%和61.46%）和HBsAg的抑制率（分别为30.17%、44.00%和57.76%）有显著变化（P<0.01），抗基因LNA组与其他四个对照组之间有显著差异（P<0.05）。抗基因LNA组中，HBV S基因的相对mRNA表达为0.33，HBsAg阳性肝细胞百分比为31%，与对照组的这两个指标有显著差异（P<0.05）。肝/肾生化参数及HE染色结果无异常变化。靶向HBV S基因的抗基因LNA在转基因小鼠中具有较强的抗病毒作用，为HBV的基因治疗提供了理论和实验依据。
