亚砷酸钠对角质形成细胞的氧化损伤作用
Oxidative injury induced by sodium arsenite in HaCaT cells
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摘要:目的 探讨亚砷酸钠(NaAsO2)对体外培养的人皮肤角质形成细胞株(HaCaT)的氧化损伤作用.方法 用Alamarblue还原法检测细胞增殖力, 以Alamarblue还原率表示细胞增殖力; 用2'7'-二乙酰二氯荧光素(DCFHDA)测定细胞内活性氧群(ROS)水平; 用彗星实验检测DNA损伤; 用紫外速率直接法测定细胞内过氧化氢酶(CAT)的活性.结果 NaAsO2作用于HaCaT细胞24h后, 0.001~1μmol/L组的AlamarBlue还原率显著高于对照组, 而10μmol/L以上组AlamarBlue还原率下降(P < 0.05);25μmol/L的NaAsO2即可使二氯荧光素(DCF)的荧光强度增加; 5μmol/L的NaAsO2可引起单个细胞的彗星尾长显著高于对照组; 25μmol/L的NaAsO2即可引起CAT活性明显降低(P < 0.05), 且呈剂量-反应关系.结论 低浓度NaAsO2可刺激细胞增殖而高浓度的NaAsO2抑制细胞增殖, NaAsO2可引起HaCaT细胞内ROS增多, 引起细胞DNA损伤和CAT活性降低.Abstract:Objective Tolearn the oxidative injury induced by sodium arsenite in HaCa Tcells.Methods AlamarBlue assay was used to evaluate the proliferation of HaCa T.Reactive oxygen species(ROS)was detected by staining cells with DCFH-DA.DNA damage was evaluated by comet assay.Intracellular catalase(CAT)activity was detected with ultraviolet velocity direct method.Results The reduction of AlamarBlue increased significantly when the sodium arsenite were less than 10μmol/L(P < 0.05), while the reduction reduced significantly when the concentration was above 100μmol/L(P < 0.05); Fluorescent density of dichlorofluorescein(DCF)increased and CATactivity decreased significantly in all groups treated with sodium arsenite while comet tail was longer when sodium arsenite were higher than 5μmol/L(P < 0.05).Conclusion Low levels of sodium arsenite can stimulate the proliferation of HaCa Twhile high levels inhibit it.Arsenic can induce enhancive ROS, and also DNA damage and inhibition of CATactivity.
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