The Study Of Toxicity On Arsanilic Acid Induced Apoptosis Of Rat Hepatocyte In Primary Culture

With the extensive use in addition agent of multipara, the arsanilic acid has resulted in remnant in products and environment pollution. So we carry out this research.Research Aim: dicuss toxicity trauma and function mechanism of arsanilic acid on cultured hepatocyte in vitro, and find minimum concentration range of arsanilic acid that can induce apoptosis of hepatocyte so that can provide theoretic basis on arsanilic acid's additive standard and estimate of arsanilic acid's feed sanitation and food safety.Research Methods: rat primary hepatocyte is used as empirical model. Rat hepatocytes were isolated by a single tow-step perfusion method. All the rat hepatocytes were treated with arsanilic acid at different mass concentrations (0, 5, 50, 500 u mol/L and 5, 10 mmol/L). Injury of hepatocytes was detected by MTT assay. Then apoptosis was observed by DNA agarose gel lelectrophoresls. The activity of SOD in hepatocytes and cell culture solution was determined by xanthine oxidase assay, and measured the git-up of GSH-Px and changes of H2O2 in cell culture solution. The changes of expression of Bax was determined by immunohistochemical method.Research Results: persistence of isolated hepatocytes was between 85% to 95%. The hepatocytes growing was inhibited by 50 U mol/L arsanilic acid in dose-dependent and time-dependent manners. And DNA Ladder was detected at 24h and 48h, that meaned the hepatocytes was induced by arsanilic acid. The git-up of SOD in cell culture solution was reduced by 50~500 u mol/L arsanilic acid. The cntent of H2O2 in culture solution was heightened by 500 U mol/L arsanilic acid after 24h. The git-up of SOD and GSH-Px in hepatocytes were reduced by 5 u mol/L and 500 u mol/L arsanilic acid respectively. The expression of Bax was boosted up by arsanilic acid.Research Conclusion: arsanilic acid effectively inhibits the proliferation of rat hepatocytes in primary culture and induced apoptosis. The restrain of SOD and GSH-Px and elevation of Bax expression were the mechanism of arsanilic acid inducing apoptosis. The minimum of concentration range of arsanilic acid inducing hepatocytes apoptosis was between 50 to 500 u mol/L.