芥子气对人皮肤角质细胞DNA损伤效应的研究

发布时间：2018-06-20 10:05

  本文选题：芥子气 + 人皮肤角质细胞　； 参考：《军事医学》2017年08期

【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(P0.05),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。
[Abstract]:Objective to explore the toxic effects of mustard gas on the DNA damage of human immortalized epidermal keratinocytes (HacaT cells) and its mechanism. Methods the inhibitory effect of mustard gas on the proliferation of HacaT cells was detected by CCK-8 method. The effect of mustard gas on the apoptosis index of HacaT cells was detected by Annexin V -FITC method, and single cell gel electrophoresis was used to detect mustard. The effects of seed gas on HacaT cell DNA and the effect of mustard gas on DNA damage and repair related protein in HacaT cells were detected by Western blot. Results the proliferation of HacaT cells with different concentrations of mustard gas was inhibited (P0.05) after 24 h of HacaT cells, and showed a significant dose dependence. The median inhibitory concentration was 121 Mu and HacaT cells. Apoptosis index and comet trailing rate increased with the increase of mustard gas concentration, and different concentrations of mustard gas could lead to changes in the expression and activity of DNA damage repair related proteins. Conclusion mustard gas has obvious cytotoxic effect on HacaT cells, which can cause apoptosis and produce obvious DNA damage, and ATM-P53- gamma H2AX-PARP signals pass through. The pathway plays an important role in the DNA damage repair process induced by mustard gas.
【作者单位】： 解放军疾病预防控制所;军事医学科学院毒物药物研究所;
【基金】：全军后勤科研重大项目资助项目(AWS11C004)
【分类号】：R99
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本文编号：2043887