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 ?mol?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.
ABSTRACT
AIM: To study the inhibitory effects of 10-23 deoxyribozyme (10-23 DRz) on Escherichia coli ?-lactamase gene expression. METHODS: According to the gene sequence of Escherichia coli ?-lactamase gene blashv-5, 10-23 DRz and antisense oligonucleotides (As-ODN) were designed and synthesized. 10-23 DRz, As-ODN or control oligonucleotides were respectively introduced into Escherichia coli by the method of electroporation. Following electroporation, bacterial viability in liquid medium contained ceftazidime was detected, bacterial ?-lactamase expression was analysed by using IEF-PAGE and the ?-lactamase band was measured with gel documentation-analyzing system. RESULTS: A_ 600 in 10-23 DRz transfected Escherichia coli was lower compared with that in As-ODN transfected Escherichia coli (P