In Vivo Studies On Hepatitis B Virus Inhibition By Using Lentiviral Vector Mediated Small Interference RNA Against HBV

Objective: To develop a mouse model of acute hepatitis B virus infection and observe the inhibition of HBV replication and expression by using Lentiviral Vectors(LVs) mediated small interfering RNA targeting HBV. Methods: Thirty-six Balb/c mice were randomly divided into three groups(N=12,each). 1.Negative control group(NC group):the model of acute hepatitis B virus infection were established by administrating pTHBV2(the eukaryotic expression vectors which contained HBV genome informations) through tail-vein hydrodynamic injection. 2. Experimental group(E group): pTHBV2 and pWPT-HBV-siRNA1(the lentivirus mediated siRNA targeting HBV HBsAg) were administrated through tail-vein. 3. control group(I group): pTHBV2 and pWPT-control-siRNA(unrelated HBV siRNA) were administrated by using the same method. The sera and liver tissues were collected on the fourth and seventh days respectively. ELISA method was used to detect HBV surface antigen(HBsAg) in serum; RT-PCR was performed to detect HBV S-mRNA level in hepatic tissue. Intracellular HBV core antigen (HBcAg) and HBsAg were determined by immunohistochemistry. Results: In contrast with NC group, serum HBsAg level in E group was decreased by 89.76% and 94.81% on the fourth and the seventh days respectively (P<0.01);Liver tissue HBV S-mRNA level, intracellular HBcAg and HBsAg levels were also suppressed significantly (P<0.05). However, there was no significantly different regarding serum HBsAg, tissue HBV S-mRNA,intracellular HBcAg and HBsAg levels between NC group and I group(P>0.05). Conclusions: Based on the results of this experiment, Lentiviral vector mediated HBV siRNA could remarkably inhibit the replication and expression of HBV in vivo, which was augmentated from the fourth day to the seventh day.