Effect of oxymatrine on the replication cycle of hepatitis B virus in vitro.
OBJECTIVE
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 microg/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 microg/mL of oxymatrine treatment for 2 d and 5 d, respectively.


CONCLUSIONS
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.
Xu
Wen-Sheng
WS
Department of Infectious Diseases, Changzheng Hospital, Second Military Medical University, No. 415 Fengyang Road, Shanghai, China.
Zhao
Ke-Kai
KK
Miao
Xiao-Hui
XH
Ni
Wu
W
Cai
Xiong
X
Zhang
Rui-Qi
RQ
Wang
Jun-Xue
JX
D000470:Alkaloids / Q000494:pharmacology*; D000818:Animals; D000998:Antiviral Agents / Q000494:pharmacology; D001483:Base Sequence; D045744:Cell Line, Tumor; D017931:DNA Primers / Q000235:genetics; D004270:DNA, Circular / Q000235:genetics; D004305:Dose-Response Relationship, Drug; D006509:Hepatitis B / Q000821:virology*; D006515:Hepatitis B virus / Q000378:metabolism*; D006801:Humans; D051379:Mice; D008822:Mice, Transgenic; D008969:Molecular Sequence Data; D011807:Quinolizines / Q000494:pharmacology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D000093842:Matrines
D000470:Alkaloids; D000998:Antiviral Agents; D017931:DNA Primers; D004270:DNA, Circular; D011807:Quinolizines; C037573:oxymatrine; D000093842:Matrines