Screening of Inhibition of Human Hepatitis B Virus Replication by Momordica Charantia Compounds

• Main Authors: Bo-Wei Lin, 林柏瑋
• Other Authors: Wen-Ling Shin
• Format: Others
• Language: zh-TW
• Published: 2010
• Online Access: http://ndltd.ncl.edu.tw/handle/25567557262852741465

碩士 === 國立屏東科技大學 === 生物科技研究所 === 98 === Hepatitis B virus（HBV）infection is a global health problem and may cause acute, fulminant and chronic hepatitis. Ths risk of chronic hepatitis B carriers, leading to cirrhosis and even hepatocelullar carcinoma（HCC）is higher than non-HBV carriers about 200-fold. In Taiwan, more than 90％ of people have been infected by HBV, more than 15％ of whom develop into chronic carriers. Therefore, HBV is still a significant problem and the development of new drugs against hepatitis B is imperative. 2.2.15 is a hepatoblastoma cell line, abidingly secretion of hepatitis B virus particles and is a convenient platform for study of hepatitis B virus replication and screening of anti-HBV compounds. Previous studies had revealed that Momordical charantia compounds, possess anti-diabetic, anti-cancer, anti-virus and immunomodulatory effects. Therefore, we investigated whether Momordical charantia compounds could inhibit the HBV replication by using 2.2.15 cell cultured systems. Cells were treated with different concentrations of nine Momordical charantia compounds and then performed MTT assay to determine the cytotoxicity. The following experiments were performed under the IC20 concentrations. The HBsAg and HBeAg levels in conditioned medium were quantified by using the enzyme-linked immunosorbent assay. Results indicated RA2-19 and RA2-10 compounds could inhibit the production of the HBsAg and HBeAg. The inhibitiory ability is comparable to the anti-HBV drugs, oltipraz. Semi-quantitative PCR analysis, RA2-19 and RA2-10 compounds also could inhibit the production of the HBV DNA. RA2-19 and RA2-10 compounds treatment induced the elevated expression of p53 protein and phosphorylation of p53 at certain serine residues. In addition, RA2-19 and RA2-10 increase the p53 transcriptional activation activities. Over-expression of p53 dominant negative mutant released the RA2-19 and RA2-10-mediated the p53 transcriptional activation function. Takentogether, RA2-19 and RA2-10 might utilize the p53 pathways to exert their anti-HBV activity, which is similar to oltipraz. In an animal model, RA2-10 significantly prevented tumor formation comparably with silymarin. RA2-19 and RA2-10 reduced the production of HBsAg and HBeAg under the non-toxic concentration, suggesting these two compounds can be further developed as anti-virus and even anti-liver cancer new drug/ health food.