反义锁核酸在转基因小鼠体内阻断乙肝病毒C基因表达

摘要

Objective To investigate the inhibitory effects on hepatitis B virus (HBV) replication of antisense locked nucleic acid (LNA) targeting to C gene in transgenic mice.Methods Antisense LNA were injected into transgenic mice via the tail vein.Serum HBV DNA was tested by real-time PCR.Serum HBeAg was tested by timeresolved fluorescence immune assay.The expression of HBcAg in the liver was detected by immuneohistochemistry.Results Five days after LNA injection,serum HBV DNA level in the AS-LNA group was reduced by 44.47％,and serum HBeAg level was decreased by 63.46％.These values were significantly higher than those in the control groups (all P ＜ 0.05).The expression of HBcAg in liver was significantly lower than that in control groups.Conclusions Antisense LNA targeting to C gene has strong inhibitiory effect on HBV replication and expression in transgenic mice and C gene is a potentical target as gene therapy.%目的 探讨针对乙肝病毒C基因反义锁核酸在转基因小鼠体内阻断病毒基因表达的效果.方法 各实验组经尾静脉给药后,采用real-time PCR、时间分辨免疫荧光技术、免疫组织化学法等技术分别检测血清HBV DNA、HBeAg含量及肝细胞内HBcAg的表达情况.结果 注射锁核酸后,对乙肝病毒核酸的复制和乙肝病毒e抗原的合成均有较强的抑制作用,注射后1、3 和5 d,HBV DNA的平均抑制率分别19.24％、39.20％和44.47％；HBeAg的平均抑制分别为30.71％、51.14％和63.46％；小鼠肝细胞的HBcAg阳性细胞数也较对照组明显减少.结论 针对乙肝病毒C基因的反义锁核酸在转基因小鼠体内能有效抑制病毒基因的复制和表达,提示C基因可作核酸药物治疗的有效靶位.