Experimental Study Of Inhibition Of Hepatitis B By Dual-target Antisense RNA In HBV Transgenic Mice

Objective To investigate the effect of dual-target antisense RNA of hepatitis B virus (HBV) targeted to X and P region on replication and expression of HBV gene in HBV transgenic mice.Methods Recombinant vector plasmids expressing dual-target antisense RNA complementary to HBV X and P region were injected into transgenic mice through tail vein. The experimental mice were divided into blank, PLXSN, PLXSN -asX, PLXSN -asP, PLXSN -asXP and PLXSN -seXP groups. The HBsAg in serum were tested by ELISA method. HBV DNA in serum were tested by FQ-PCR. HBsAg and HBcAg in cell of heart, liver, spleen, lung and kidney were tested by immunohistochemistry method. Side effect was tested by pathological section.Results PLXSN-asX and PLXSN-asXP treatment of the mice for 4 weeks could initiate an decrease by 24% and 27% respectively in the expression level of HbsAg. Though, no significant change in other groups was observed. The expression of HBV DNA in serum were inhibited distinctivsely(58%) in PLXSN-asX group at 2nd week while in PLXSN-asP group at 8th week, and in PLXSN-asXP group the inhibitory effect on the expression of HBV DNA reached 66% at 1st week and 77% at 8th week after treatment, as in other groups not. The expression of HBsAg and HBcAg in hepatocyte of PLXSN-asX group, PLXSN-asP group and PLXSN-asXP group were lower than other groups after 8-week treatment. There is no expression of HBsAg and HBcAg inheart, spleen and lung in any mice. But the expression of HBcAg in kidney were found in mice except PLXSN-asXP group. No significant abnormality was found in the tissues in all groups.Conclusion HBV antisense RNA can inhibit the replication and expression of HBV gene in HBV transgenic mice. The inhibitory effect of dual-target antisense RNA is higher than that of single-target antisense RNA.