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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。
Objective To investigate the toxicity of different inorganic arsenic and organic arsenic compounds on Chang hepatocytes and the induction of Ⅱ phase detoxification enzyme and antioxidant enzyme protein. Methods The cell viability was determined by Alamar Blue reduction method. The protein expression of HO-1, NQO1, GST and GR was detected by Western blot. Results The proliferation activity of the cells was determined by Alamar Blue reduction method. The toxicity of several arsenic compounds was As2O3> NaAsO2> Na2HAsO4> DMA. After treated with different concentrations of inorganic arsenic for 6 hours, High, HO-1 and NQO1 protein expression levels; GST and GR protein expression and concentration has nothing to do, all three groups were expressed and the same; and organic arsenic DMA is the opposite, as the concentration increased, GST and GR protein expression levels gradually increased, HO-1, NQO1 protein expression level has nothing to do with the exposure concentration, the exposure group compared with the control group no difference. Conclusion The toxicity of different arsenic compounds to hepatocytes is different. For the protein expression of anti-oxidase and phase Ⅱ detoxification enzymes, the targeted and induced intensities of the four arsenic compounds are obviously different.