Preliminary Study On The Mechanism Of Hsa＿circ＿0000038 Regulating HBV Replication In Human Chimeric Liver Mice

Objectives:1.To investigate whether hsa＿circ＿0000038 regulates the replication of hepatitis B virus(HBV)in humanized chimeric liver mice.2.To explore the effect of hsa＿circ＿0000038 on the expression of nuclear factor 1A(NFIA)and histone demethylase(JMJD3).Methods:1.The viability of the humanized chimeric liver mice was confirmed using serum human albumin assay kits and immunofluorescence of liver tissue sections.2.To create an HBV-infected mouse model,all healthy humanized chimeric liver mice were infected with serum from untreated chronic hepatitis B patients.On this basis,the mice were reinfected with adeno-associated virus 9(AAV9)expressing hsa＿circ＿0000038 gene or empty vector respectively,resulting in the construction of hsa＿circ＿0000038 overexpression mouse model or empty control mouse model.3.The expression levels of HBV DNA,pre-genomic RNA(pg RNA)and ccc DNA in mouse model were quantified by real-time quantitative polymerase chain reaction(RTqPCR).4.RT-qPCR and Western blot methods were used to detect the expression levels of JMJD3 and NFIA in the treated liver tissues.Result:1.Human serum albumin(HSA)was detected in the serum of both experimental and control mice,and specific fluorescence of Human cytokeratin-18(CK-18)was detected in the liver tissue sections.2.After mice were infected with HBV,HBV DNA was detected in the serum of both experimental and control mice,and the level of HBV DNA tended to increase continuously.3.The m RNA expression level of hsa＿circ＿0000038 in the liver tissue of the experimental mice was significantly increased compared to the control group at week6 after infection with recombinant adeno-associated virus(AAV9-circ＿0000038/AAV9-control).4.Overexpression of hsa＿circ＿0000038 resulted in significant upregulation of HBV DNA,HBs Ag and HBe Ag levels in serum and ccc DNA levels in liver tissues of experimental mice.5.Overexpression of hsa＿circ＿0000038 significantly upregulated the expression levels of NFIA and JMJD3 in the liver of experimental mice.Conclusion:1.hsa＿circ＿0000038 promotes HBV replication in humanized chimeric liver mice.2.hsa＿circ＿0000038 contributes to the transcription and accumulation of HBV ccc DNA.3.hsa＿circ＿0000038 promotes the expression of NFIA and JMJD3 m RNA.