无机砷化合物对HaCaT细胞毒性作用  

Cytotoxic effects of different kinds of inorganic arsenicals on HaCaT cells

作　　者：陆景坤[1] 田艳[1] 陈朝军[1] 俞腾飞[1] 王一博[1] 李中燕[1] 

机构地区：[1]内蒙古医科大学蒙医药研究院GLP中心,内蒙古呼和浩特010010

出　　处：《中国公共卫生》2013年第3期387-389,共3页Chinese Journal of Public Health

基　　金：内蒙古教育厅基金资助项目(NJ09103)

摘　　要：目的探讨无机砷化合物对人表皮角质细胞(HaCaT)毒性与氧化应激和砷代谢关系。方法用四甲基偶氮唑盐法测定细胞生存率;检测砷化物对丙二醛(MDA)含量、超氧化物歧化酶(SOD)活力影响;流式细胞仪检测细胞内活性氧(ROS);原子荧光分析方法检测细胞内外不同形态砷含量。结果 200.0μmol/L As2O3(iAsⅢ)、50.0～500.0μmol/L Na2HAsO4.7H2O(iAsⅤ)能够明显降低HaCaT细胞生存率,生存率分别为(54.8±5.9)%、(85.4±9.5)%、(70.0±4.0)%、(58.8±6.0)%,在低剂量时,2种砷化物表现为促进HaCaT细胞增殖(P<0.05);iAsⅢ组随剂量增加MDA含量梯度增加,iAsⅤ组MDA含量先增加后降低呈倒U形;细胞内ROS含量iAsⅢ组>iAsⅤ组;细胞外iAsⅢ、iAsⅤ含量均高于细胞内,细胞内砷甲基化率表现为5.0 mol/L iAsⅢ组(87.3%)>50.0 mol/L iAsⅢ组(61.8%),5.0 mol/L iAsⅤ组(40.0%)>50.0 mol/L iAsⅤ组(10.5%),且iAsⅢ组高于iAsⅤ组。结论无机砷化物对HaCaT细胞毒性影响与氧化应激和砷代谢水平有关。Objective To investigate the relationship between cytotoxicity and oxidative stress and arsenic metabo- lism induced by different kinds of inorganic arsenicals in epidermal keratinocytes cells （HaCaT）. Methods Methyl thia- zolyl tetrazolium（MTT） assay was used to evaluate the cell viability. Malondialdehyde（MDA） content and the activity of the superoxide dismutase （SOD） in HaCaT were detected. The intracellular reactive oxygen species （ROS） were detected with flow cytometry and the atomic fluorescence was used to analyze intracellular or extracellular level of different forms of arsenic. Results As2 03 （200.0 μmol/L） and Na2 HAsO4 · 7H2O （ ≥ 50. 0 μmol/L） could decrease the cell viability, with the survival rates of 54. 8 ± 5.9% and 85.4 ± 9.5%, 70. 0 ± 4. 0% and 58.8± 6. 0%, and could significantly promote the proliferation of HaCaT cell at low dosage（ P 〈 0. 01 ,P 〈 0. 05 ）. The MDA levels of iAsm groups increased with dose increasing and iAs v groups showed decrease after an increase with an U-shaped dose-respose curve. The SOD activity showed a dose-effect relation with an U-shaped curve. The intracellular ROS decreased with the order of As203 , N% HAsO4 · 7H2 O. The extracellular iAs m, iAs v contents were higher than intracellular contents in all groups. The meth- ylation ratio of 5.0 mol/L iAsm group（87. 3% ） was higher than that of 50.0 mol/L iAs （61.8%） and the ratio of 5.0 mol/L iAsv groups（40. 0% ） was higher than that of 250.0 mol/L iAsv （ 10. 5% ）. The ratio of iAsm groups was higher than that of the iAsv groups. The arsenic levels of the high-dose groups were higher than those of low-dose groups. The efflux ability of low-dose groups were higher and the efflux ability of iAsr groups were lower than those of the iAsv groups. Conclusion The mechanisms of inorganic arsenic toxicity probably relate to different levels of oxidative stress and arsenical metablism.

关 键 词：砷化合物 人表皮角质细胞(HaCaT) 氧化应激 砷代谢 

分 类 号：R387[医药卫生—医学寄生虫学]