Inhibition of hepatitis B virus S gene expression by anti-gene locked nucleic acid in vivo / 医学研究生学报

ABSTRACT

Objective There is still a lack of effective treatment for hepatitis B. The article aimed to investigate the inhibito-ry and antiviral effects of hepatitis B virus(HBV)S gene expression by anti-gene locked nucleic acid(LNA)in vivo in transgenic mice. Methods The HBV transgenic mice were divided into 5 groups by random number,6 in each group. They were blank group, irrelevant sequence group,lamivudine group,antisense locked nucleic acid group,and anti-gene locked nucleic acid group respective-ly. The lamivudine group was treated by oral gastric lavage,with 2mg/kg dose 2 times per day for continuous 7d,and the rest groups were injected with 500 mL by tail vein injection at 1,3,5 d after ad-ministration. HBV DNA was tested by real-time quantitative polymer-ase chain reaction(qPCR);HBsAg was tested by ELISA;mRNA of HBV S gene was detected by reverse transcription PCR(RT-PCR);the positive rate of HBsAg in liver cells was detected by immunohisto-chemistry. Results After the treatment of anti-gene locked nucleic acid on 3rd,5th,7thday,the inhibition rate of HBV DNA were 37.18%,50.27%,61.46%,and HBsAg were 30.17%,44%,57.76%. The inhibitory effect on 7thday was more obvious than those of the blank group,the unrelated sequence group,the lamivudine group and the antisense lock nucleic acid group,and the difference was statistically significant(P<0.05).The expression of mRNA S gene HBV(0.33)and liver cells HBsAg positive rate(31%)was signif-icantly reduced compared with control group(P<0.05).No abnormal change was found in the function of liver and kidney tests and tis-sue HE staining. Conclusion Anti-gene lock nucleic acid targeting to S gene has strong inhibitory effects on HBV replication and expression in transgenic mice,which provides theoretical and experimental knowledge for HBV gene therapy.