Synergistic effect of a novel oxymatrine-baicalin combination against hepatitis B virus replication, alpha smooth muscle actin expression and type I collagen synthesis in vitro.
OBJECTIVE
To study the effect of oxymatrine-baicalin combination (OB) against HBV replication in 2.2.15 cells and alpha smooth muscle actin (alpha SMA) expression, type I, collagen synthesis in HSC-T6 cells.


METHODS
The 2.2.15 cells and HSC-T6 cells were cultured and treated respectively. HBsAg and HBeAg in the culture supernatants were detected by ELISA and HBV DNA levels were determined by fluorescence quantitative PCR. Total RNA was extracted from HSC-T6 cells and reverse transcribed into cDNA. The cDNAs were amplified by PCR and the quantities were expressed in proportion to beta actin. The total cellular proteins extracted from HSC-T6 cells were separated by electrophoresis. Resolved proteins were electrophoretically transferred to nitrocellulose membrane. Protein bands were revealed and the quantities were corrected by beta actin.


RESULTS
In the 2.2.15 cell culture system, the inhibitory rate against secretion of HBsAg and HBeAg in the OB group was significantly stronger than that in the oxymatrine group (HBsAg, P = 0.043; HBeAg, P = 0.026; respectively); HBV DNA level in the OB group was significantly lower than that in the oxymatrine group (P = 0.041). In HSC-T6 cells the mRNA and protein expression levels of alpha SMA in the OB group were significantly lower as compared with those in the oxymatrine group (mRNA, P = 0.013; protein, P = 0.042; respectively); The mRNA and protein expression levels of type I collagen in the OB group were significantly lower as compared with those in the oxymatrine group (mRNA, P < 0.01; protein, P < 0.01; respectively).


CONCLUSIONS
OB combination has a better effect against HBV replication in 2.2.15 cells and is more effective against alpha SMA expression and type I collagen synthesis in HSC-T6 cells than oxymatrine in vitro.
Cheng
Yang
Y
Institute of Liver Disease, Shuguang Hospital affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China. yangcheng@myrealbox.com
Ping
Jian
J
Xu
Huai-Dong
HD
Fu
Hai-Jun
HJ
Zhou
Zhao-Hui
ZH
D000199:Actins / Q000378:metabolism*; D000470:Alkaloids / Q000008:administration & dosage*; D000890:Anti-Infective Agents / Q000008:administration & dosage*; D000998:Antiviral Agents / Q000008:administration & dosage*; D024042:Collagen Type I / Q000096:biosynthesis*; D004357:Drug Synergism; D004359:Drug Therapy, Combination; D004797:Enzyme-Linked Immunosorbent Assay; D005419:Flavonoids / Q000008:administration & dosage*; D006515:Hepatitis B virus / Q000187:drug effects / Q000378:metabolism*; D019694:Hepatitis B, Chronic / Q000188:drug therapy*; D006801:Humans; D008099:Liver / Q000187:drug effects / Q000821:virology*; D011807:Quinolizines; D000093842:Matrines
D000199:Actins; D000470:Alkaloids; D000890:Anti-Infective Agents; D000998:Antiviral Agents; D024042:Collagen Type I; D005419:Flavonoids; D011807:Quinolizines; C038044:baicalin; C037573:oxymatrine; D000093842:Matrines