多种砷化物对Chang肝细胞抗氧化酶的诱导作用

目的探讨不同无机砷和有机砷化物对Chang肝细胞的毒性,以及对Ⅱ相解毒酶和抗氧化酶蛋白的诱导作用。方法采用Alamar Blue还原法测定细胞活力;Western blot免疫印迹法检测细胞内HO-1、NQO1、GST和GR的蛋白表达。结果通过Alamar Blue还原法测定细胞增殖活力,几种砷化物的毒性大小依次为As2O3〉NaAsO2〉Na2HAsO4〉DMA;不同浓度的无机砷染毒Chang肝细胞6 h后,发现随着染毒浓度的升高,HO-1和NQO1的蛋白表达水平逐渐增加;GST和GR的蛋白表达与染毒浓度无关,3个组都表达且相同;而有机砷DMA正相反,随着染毒浓度的升高,GST和GR的蛋白表达水平逐渐增加,HO-1、NQO1的蛋白表达水平与染毒浓度无关,染毒组与对照组相比无差异。结论不同砷化物对肝细胞的毒性不同;对于抗氧化酶和Ⅱ相解毒酶的蛋白表达,4种砷化物的针对性和诱导强度明显不同。

Induction of various arsenide on antioxidase in Chang liver cells

Objectives To explore different organic and inorganic arsenics toxicity in Chang liver cells and the induction on phase Ⅱ detoxifying enzymes and antioxidant enzyme protein. Methods Alamar Blue reduction method was used to evaluatecell viability; Western blot analysis was used to detect the protein expression levels of HO-1,NQO1, GST and GR in cells. Resuits Through the Alamar Blue determination of cell proliferation activity, we found the toxic size of As2 O3 〉 NaAsO2 〉Na2 HAsO4 〉 DMA ; Different concentrations of inorganic arsenic exposure Chang liver cells for 6h, protein expression levels of HO -1 and NQO1 gradually increased; GST and GR protein expression had nothing to do with exposure concentration, three groupsshared the same expression level. While,the organic arsenic DMA was totally opposite; as the exposure concentration elevated, GST and GR protein expression levels gradually increased, the protein expression of HO-1 and NQO1 had nothing to do with exposure concentration. Exposure group and control group had no difference. Conclusion Different arsenide had different toxic effect on the Chang liver cells. Different organic and inorganic arsenic have different induced intensity for phase Ⅱ detoxifying en-zymes and antioxidant enzymes expression.