反义锁核酸在转基因小鼠体内阻断乙肝病毒C基因表达

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.

Inhibition of hepatitis B virus(HBV) replication using antisense LNA targeting to C gene in HBV transgenetic mice

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 time-resolved fluorescence immune assay.The expression of HBcAg in the liver was detected by immuneohistochemistry. Results Five days after LNA injection,serum HBV DNA levels in the AS-LNA group were reduced by 44.47%,and serum HBeAg levels were decreased by 63.46%.These values were significantly higher than those in the control groups(all P<0.05). The expression levels of HBcAg in the liver were significantly lower than those in the control groups. Conclusion The present study proves that antisense LNA targeting to C gene has strong inhibition effect on HBV replication and expression in transgenic mice, and suggests that C gene being the effected site for gene therapy.