反义硫代寡核苷酸抗乙肝病毒作用机理的探讨

PROBING INTO THE MECHANISM OF ANTISENSE OLIGONUCLEOTIDE AGAINST HEPATITIS B VIRUS

为了探讨反义寡核苷酸（ＡＳＯＮ）进行抗乙型肝炎病毒（ＨＢＶ）基因治疗的可行性及ＡＳＯＮ抗ＨＢＶ作用机理，设计合成了针对ＨＢＶ不同基因位点的ＡＳＯＮ〔ＰｒｅＳ２翻译起始位点、ＨＢＶ调节成份顺向重复序列（ＤＲ２）和增强子Ⅱ〕。以ＨｅｐＧ２·２·１５细胞为细胞模型，动态观察了这三种ＡＳＯＮ不同浓度不同时间的抗ＨＢＶ作用。结果表明：这三种ＡＳＯＮ对ＨＢｅＡｇ的抑制率分别为９１．１％、４４９％和５６４％，对ＨＢｓＡｇ的抑制率分别为６５７％、６０１％和５１５％，无关序列无明显抑制作用。应用反义寡核苷酸技术，为研制新一代抗ＨＢＶ基因治疗药物奠定了基础，探讨了特异性阻断ＨＢＶ基因表达的可能途径。

In order to study the feasibility of gene therapy of oligonucleotide (ASON) against hepatitis B viurs, three kinds of ASON were designed and synthesized against different gene regions of HBV (translational initiation site of the pre S 2 gene,direct repeat sequence DR 2 and Enhancer Ⅱ). The 2·2·15 cells were chosen as an in vitro cell culture assay system and the inhibitory effect of the three kinds of ASON was observed consecutively at different concentration and at different time. The results showed that their inhibitory rates against HBsAg expression are 65.7%、60.1% and 51.5% respectively,and against HBeAg expression, 91.4 %、44.9% and 56.4% respectively. There was no inhibitory effect on the expression of viral protein with an unrelated oligonucleotide(<12%). The study provides a foundation for the development of a new generation of gene therapy drugs against HBV and probes into the possible ways to spectific blocking HBV gene expression by using antisense oligonucleotides. The mechanism of ASON against HBV is possibly its effect on the level of reverse transcription and translation or replication.