Inhibition on Hepatitis B virus in vitro of recombinant MAP30 from bitter melon.
The gene encoding MAP30 protein was cloned from bitter melon and recombinant MAP30 was expressed and purified. The human hepatoma G2.2.15 cells were exposed to different concentrations of MAP30. MTT assay was used to evaluate the cytotoxicity of the drugs and real-time PCR and Southern hybridization were applied to quantify extracellular HBV DNA and replicative intermediates intracellular and cccDNA in nucleus. HBsAg and HBeAg were assessed by enzyme-linked immunosorbent assay (ELISA). The results showed that exposure of HepG2.2.15 cells to MAP30 resulted in inhibition of HBV DNA replication and HBsAg secretion. After exposed to three different concentrations of MAP30 for 2, 4, 6, and 8 days respectively, the inhibition rates of extracellular HBV DNA, HBsAg, and HBeAg of each concentration decreased significantly (P < 0.05). After 9 days of treatment, the inhibition rates of extracellular HBV DNA of the different concentrations differed greatly (P < 0.001). The MAP30 could inhibit the production of HBV (P < 0.01) dose-dependently. The expression of HBsAg was significantly decreased by MAP30 dose-dependently (P < 0.001) and time-dependently (P < 0.001). Lower dose of MAP30 (8.0 microg/ml) could inhibit the expression of HBsAg and HBeAg.
The Laboratory of Toxicology, College of Public Health, Zhengzhou University, Zhengzhou 450001, China. 5746067@sohu.com
Jian Ming
JM
Fan
Qiao
Q
Zhang
Jun
J
Xu
Sha
S
Zhu
Tao
T
Ke
De Fu
DF
Gao
Yu Bao
YB
Xu
D000998:Antiviral Agents / Q000494:pharmacology*; D045744:Cell Line, Tumor; D004261:DNA Replication / Q000187:drug effects; D004279:DNA, Viral / Q000037:antagonists & inhibitors; D004305:Dose-Response Relationship, Drug; D006515:Hepatitis B virus / Q000187:drug effects*; D006801:Humans; D029777:Momordica charantia / Q000737:chemistry; D010940:Plant Proteins / Q000627:therapeutic use; D011994:Recombinant Proteins; D054790:Ribosome Inactivating Proteins, Type 2 / Q000302:isolation & purification / Q000494:pharmacology*
D000998:Antiviral Agents; D004279:DNA, Viral; C066347:MAP30 protein, Momordica charantia; D010940:Plant Proteins; D011994:Recombinant Proteins; D054790:Ribosome Inactivating Proteins, Type 2