反义寡核苷酸对乙型肝炎病毒的体外抗病毒作用

Inhibihon of hepatitis B viral gene expression by anti-sense Phosphrothioate oligonucleotides

以乙型肝炎病毒基因转染Ｈｅｐ－Ｇ２细胞（２、２、１５细胞）为实验对象，合成互补于乙型肝炎病毒基因ｍＲＮＡＳＰＩＩ增强子区及Ｓ基因翻译起始区的１５－聚反义硫代磷酸脱氧核苷；通过检测作用细胞上清液中ＨＢｓＡｇ、ＨＢｅＡｇ的分泌情况，证明上述２段反义寡核苷酸在１．Ｏ～５．０ｕｍｏｌ／Ｌ浓度范围内可剂量相关性地抑制乙型肝炎病毒基因表达，继续增大用药剂量则出现抑制平台现象，而对照序列则无抑制作用。反义寡核苷酸可特异性地抑制７８％～８６％ＨＢｓＡｇ及４０％～５５％ＨＢｅＡｇ的产生，而对细胞自身蛋白合成无影响，亦未观察到细胞毒害作用。个研究为从基因水平探讨研制新一代抗乙肝病毒药物提供了线索。

ep G2 cells transfected with HBV genomes wereused to study the inhibitory effect of antisensephosphrothiate oligodeoxynuclcotides on HBsAg andHBeAg production. The synthetic antisense15-s-oligomers against the cap site of mRNA tran-scribed from the SPII promoter and regions of thetranslational initiation site of the S gene showed adose-dependent and sequence-specific inhibitory ef-fect on HBV gene expression between concentrationsof l.0 ￣ 5.0 μmol / L. But the oligomers directed onthe middle site of the S gene exerted little effect onHBsAg and HBeAg expression as well as thenon-complementary random sequence control. Thecells remained viable throughout the experiments andno moophological abnonnalities and cytotoxicity wereobserved with antisense S-oligoniers at concentrationsbelow 20.0μmol/L. These results suggest atherapeutic potential for antisense oligonucleotides inthe treatment of patients who are chronically infectedwith HBV.