Brief Article
Copyright ©2010 Baishideng. All rights reserved.
World J Gastroenterol. Apr 28, 2010; 16(16): 2028-2037
Published online Apr 28, 2010. doi: 10.3748/wjg.v16.i16.2028
Effect of oxymatrine on the replication cycle of hepatitis B virus in vitro
Wen-Sheng Xu, Ke-Kai Zhao, Xiao-Hui Miao, Wu Ni, Xiong Cai, Rui-Qi Zhang, Jun-Xue Wang
Wen-Sheng Xu, Xiao-Hui Miao, Wu Ni, Xiong Cai, Rui-Qi Zhang, Jun-Xue Wang, Department of Infectious Diseases, Changzheng Hospital, Second Military Medical University, Shanghai 200003, China
Ke-Kai Zhao, Department of Infectious Diseases, No. 404 Hospital, Weihai 264200, Shandong Province, China
Author contributions: Xu WS and Zhao KK contributed equally to this work; Xu WS, Zhao KK, Miao XH, Ni W, Cai X, Zhang RQ and Wang JX designed the research; Xu WS and Zhao KK performed the research and preliminary analysis of the data; Zhao KK, Miao XH, Ni W, Cai X, Zhang RQ and Wang JX analyzed and compiled the data; Zhao KK wrote the paper and Miao XH revised it.
Supported by The National Natural Scientific Foundation of China, No. 30070958; and The National Key Technologies Research and Development Program of China during the 11th Five-year Plan Period, No. 2008zx1002-006
Correspondence to: Xiao-Hui Miao, MD, PhD, Professor, Department of Infectious Diseases, Changzheng Hospital, Second Military Medical University, No. 415, Fengyang Road, Shanghai 200003, China. xhmiaosh@163.com
Telephone: +86-21-81885003 Fax: +86-21-63520041
Received: December 7, 2009
Revised: January 7, 2010
Accepted: January 14, 2010
Published online: April 28, 2010
Abstract

AIM: To determine the antiviral mechanism or target of oxymatrine against hepatitis B virus (HBV).

METHODS: HepG2.2.15 cells were incubated with culture medium containing 500 μg/mL of oxymatrine for 2 and 5 d. The surface antigen of HBV (HBsAg) and e antigen of HBV (HBeAg) in supernatant were determined by ELISA. HBV DNA in supernatant, and intracellular covalently closed circular DNA (cccDNA), relaxed circular DNA (rcDNA) and pregenomic RNA (pgRNA) were quantified by specific real-time polymerase chain reaction (PCR) or reverse transcription (RT)-PCR.

RESULTS: Treatment with oxymatrine for 2 d and 5 d reduced the production of HBV by the cell line, as indicated by the decline of HBsAg (22.67%, t = 5.439, P = 0.0322 and 22.39%, t = 5.376, P = 0.0329, respectively), HBeAg (55.34%, t = 9.859, P = 0.0101 and 43.97%, t = 14.080, P = 0.0050) and HBV DNA (40.75%, t = 4.570, P = 0.0447 and 75.32%, t = 14.460, P = 0.0047) in the supernatant. Intracellular cccDNA was also markedly reduced by 63.98% (t = 6.152, P = 0.0254) and 80.83% (t = 10.270, P = 0.0093), and intracellular rcDNA by 34.35% (t = 4.776, P = 0.0413) and 39.24% (t = 10.050, P = 0.0097). In contrast, intracellular pgRNA increased by 6.90-fold (t = 8.941, P = 0.0123) and 3.18-fold (t = 7.432, P = 0.0176) after 500 μg/mL of oxymatrine treatment for 2 d and 5 d, respectively.

CONCLUSION: Oxymatrine may inhibit the replication of HBV by interfering with the process of packaging pgRNA into the nucleocapsid, or inhibiting the activity of the viral DNA polymerase.

Keywords: Oxymatrine; Hepatitis B virus; Replication intermediates; Covalently closed circular DNA; Pregenomic RNA