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Issue:ISSN 1000-7083
          CN 51-1193/Q
Director:Sichuan Association for Science and Technology
Sponsored by:Sichuan Society of Zoologists; Chengdu Giant Panda Breeding Research Foundation; Sichuan Association of Wildlife Conservation; Sichuan University
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Your Position :Home->Past Journals Catalog->2015 Vol.34 No.2

Establishment of Hepatitis B Virus Chronic Infection Mouse Model Induced by HBV cccDNA
Author of the article:YAN Lei1, ZHAO Tingting1, WANG Huijuan1, LI Xiaosong2, HUANG Ailong2, TAN Yi1, LAI Guoqi1*
Author's Workplace:1. Laboratory Animal Center, Chongqing Medical University, Chongqing 400016, China;
2. Key Laboratory of Molecular Biology on Infection Diseases and Institute for Viral Hepatitis, Chongqing Medical University, Chongqing 400016, China
Key Words:hepatitis B virus; HBV cccDNA; hydrodynamic; mouse model
Abstract:Objective To establish the mouse model of chronic viral hepatitis by hydrodynamic injection of HBV cccDNA in C57BL/6 mice. Methods A total of 29 C57BL/6 mice were divided into experimental group, control group and blank group, and injected with HBV cccDNA, pAAV-HBV1.2 and isotonic saline by hydrodynamic method, respectively. The serum samples and liver tissues were collected at different time points after injection. Radioimmunoassay was used to examine the levels of HBsAg and HBeAg. Real time PCR was used to determine the copy numbers of HBV DNA in serum and the liver. Immunohistochemistry was used to examine the levels of HBsAg and HBcAg in liver. HE staining was used to check the pathological changes of liver. SPSS software (version 17.0) was used to analyze the experimental data. Results The expression level of HBsAg in the experimental group showed a pattern of 4 peaks at day 3 and week 3, week 7 and week 9. A similar 4-peak expression pattern was detected in HBeAg at day 1 and week 1, week 4 and week 10. In the control groups, only 2 peaks at day 3 and week 8 were found in the expression pattern of HbsAg, and the peak value of HBeAg was observed at day 1, week 3, and week 10. No expression of HBsAg and HBeAg was detected in the blank group. As determined by real time PCR, although the levels of HBV DNA decreased with time, the copy numbers were significantly higher in the experimental group than those of control (P<0.01). Moreover, the copy numbers of HBV DNA in mice liver were significantly higher than those in the serum (P<0.01) at all time points. Positive expression of HBsAg and HBeAg in liver was revealed by immunohistochemistry in the experimental and control groups. The result of HE staining confirmed the symptoms of inflammation, fibrosis and necrosis in mice liver tissues. Conclusion HBV chronic infection model in C57BL/6 mice was successfully established by in vivo transduction with HBV cccDNA. The expression of HBV in the experimental groups was higher than that of control. This study provided a basic model for further understanding the mechanism of HBV infection and liver injury.
2015,34(2): 200-207 收稿日期:2014-6-23
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