关键词: 催化性, 锁核酸核酶, 2.2.15细胞, 肝炎病毒, 乙型, 抑制效应

Abstract: Objective To observe the suppression of HBsAg and HBeAg expression by DNAzyme and LNAzyme located at HBV pre-area of HBV. Methods Eecoding sequence of 10-23 DNAzyme thiolmodificated 10-23DNAzyme and LNAzyme that were directed against Pre C/C region of HBV were designed and synthesized. Experimental groups and control groups were set up. The experimental groups included 10-23 DNAzyme group,S-10-23 DNAzyme group and LNAzyme group. The control groups include blank control group, simple lipofectamine group,simple 10-23DNAzyme group and random 10-23 DNAzyme group. In the dosege of 0.16,0.64,1.28,1.60,1.92 μmol·L^-1 and the time of 12,24,36,48,60,72,84 and 96 h, the suppression of HBsAg and HBeAg expression by 10-23 DNAzyme and LNAzyme in 2.2.15 cells were studied. Results The suppression of HBsAg and HBeAg expression by 10-23 DNAzyme and LNAzyme in 2.2.15 cells were significant. The inhibitory effects caused by LNAzyme was more significant than that by thiolmodified 10-23 DNAzyme whose inhibitory effects were more significant than that of 10-23 DNAzyme. The inhibitory rates of LNAzyme and 10-23 DNAzyme thiolmodification reached (91.6±8.4)%, (78.4±2.0)% on HBsAg，respectivelly and (90.1±5.2)% , (76.4±4.8)% on HBeAg. The inhibitory effects of LNAzyme and thiolmodification of 10-23 DNAzyme were found 12 h after they were added to 2.2.15 cells, and optimized at 48 h, effective inhibitory time for LNAzyme was 84 h,for thiolmodification 10-23 DNAzyme was 72 h. Addition of LNAzyme and 10-23 DNAzyme to 2.2.15 cells didn′t exert cytotoxicity. Conclusion 10-23 DNAzyme and LNAzyme have demonstrated significant inhibitory effects on the HBsAg and HBeAg expressions in 2.2.15 cells. Morever, the inhibitory effects of LNAzyme is more significant than that of DNAzyme. LNAzyme is a specific anti-HBV therapeutic agent.

Key words: catalytic, LNAzyme, 2.2.15 cells, hepatitis B virus, inhibitory effect