芥子气对人皮肤角质细胞DNA损伤效应的研究  

DNA damage effects on human skin keratinocytes induced by sulphur mustard

作　　者：束玉磊 何俊 闫夏贝 李漾 周维 张丽 赵君 赵增明 谢剑炜[2] 彭双清 

机构地区：[1]解放军疾病预防控制所,北京100071 [2]军事医学科学院毒物药物研究所,北京100850

出　　处：《军事医学》2017年第8期647-651,共5页Military Medical Sciences

基　　金：全军后勤科研重大项目资助项目(AWS11C004)

摘　　要：目的探究芥子气对人永生化表皮角质形成细胞(HacaT细胞)DNA损伤的毒性作用特征及机制。方法采用CCK-8法检测不同浓度芥子气对HacaT细胞增殖的抑制作用,AnnexinⅤ-FITC法检测芥子气对HacaT细胞凋亡指数的影响,单细胞凝胶电泳法检测芥子气对HacaT细胞DNA的损伤,Western印迹法检测芥子气对HacaT细胞DNA损伤修复相关蛋白的影响。结果不同浓度芥子气作用HacaT细胞24 h后,对细胞的增殖均有抑制作用(P<0.05),并呈明显的剂量依赖性,其半数抑制浓度IC50值为121μmol/L;HacaT细胞的凋亡指数和彗星拖尾率随芥子气浓度的增加而增加;不同浓度芥子气染毒可导致DNA损伤修复相关蛋白表达量及活性的改变。结论芥子气对HacaT细胞具有明显的细胞毒性作用,可引起细胞凋亡并产生明显的DNA损伤,ATM-P53-γH2AX-PARP信号通路在芥子气导致DNA损伤修复过程中发挥重要作用。Objective To investigate the toxic effects and mechanisms of sulphur mustard on DNA damage of human immortalized epidermal keratinocytes（ Haca T cells）. Methods The inhibitory effect of sulphur mustard on the proliferation of Haca T cells was detected by CCK-8 method. The apoptosis index of cells was measured by Annexin Ⅴ-FITC method. The effects of sulphur mustard on DNA damage were detected by single cell gel electrophoresis. The expression levels of DNA damage and repair related proteins were detected by Western blot. Results The proliferation rate of Haca T cells was inhibited in a dose-dependent manner after treatment with sulphur mustard for 24 h（ IC50 value was 121 mol/L）.The apoptotic and comet tailing rates of cells treated with sulphur mustard also increased in a dose-dependent manner. The expression levels of DNA damage and repair related proteins were changed after treatment with sulphur mustard. Conclusion Sulphur mustard has significant cytotoxic effect on Haca T cells,and can induce apoptosis and DNA damage. In addition,ATM-P53-γH2 AX-PARP signaling pathway plays an important role in the repair of DNA damage induced by sulphur mustard.

关 键 词：芥子气 人皮肤角质细胞 DNA损伤 

分 类 号：R99[医药卫生—毒理学]