Mechanisms of sulfur mustard analog 2-chloroethyl ethyl sulfide-induced DNA damage in skin epidermal cells and fibroblasts
Mechanisms of sulfur mustard analog 2-chloroethyl ethyl sulfide-induced DNA damage in skin epidermal cells and fibroblasts
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DOI：
10.1016/j.freeradbiomed.2011.08.020
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发表时间：
2011-12-15
影响因子：
7.4
通讯作者：
Agarwal, Rajesh
中科院分区：
文献类型：
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作者：
Inturi, Swetha;Tewari-Singh, Neera;Agarwal, Rajesh
采用小鼠皮肤表皮JB 6细胞和真皮成纤维细胞，在这里，我们研究了2-氯乙基乙基硫醚（CEES），硫芥（SM）的单功能类似物的DNA损伤的机制。CEES暴露引起H2A.X和p53磷酸化以及p53在两种细胞类型中的积累，从1小时开始，持续24小时，表明CEES的DNA损伤作用，这也通过碱性彗星试验证实和定量。CEES暴露也诱导氧化应激和氧化DNA损伤的两种细胞类型，线粒体和细胞活性氧和8-羟基脱氧鸟苷水平的增加，分别测量。在区分氧化和直接DNA损伤的研究中，用谷胱甘肽（GSH）或抗氧化剂Trolox预处理1小时显示CEES诱导的氧化应激和氧化DNA损伤减少。然而，只有GSH预处理降低CEES诱导的总DNA损伤的彗星试验，H2A.X和p53磷酸化，和总p53水平。这可能是由于通过LC-MS分析检测到GSH-CEES缀合物的形成。总之，我们的研究结果表明，CEES会导致直接和氧化性DNA损伤，这表明为了挽救SM引起的皮肤损伤，需要多效药物（或鸡尾酒），可以靶向芥末皮肤毒性的多种途径。(C)2011 Elsevier Inc. All rights reserved.
Employing mouse skin epidermal JB6 cells and dermal fibroblasts, here we examined the mechanisms of DNA damage by 2-chloroethyl ethyl sulfide (CEES), a monofunctional analog of sulfur mustard (SM). CEES exposure caused H2A.X and p53 phosphorylation as well as p53 accumulation in both cell types, starting at 1 h, that was sustained for 24 h, indicating a DNA-damaging effect of CEES, which was also confirmed and quantified by alkaline comet assay. CEES exposure also induced oxidative stress and oxidative DNA damage in both cell types, measured by an increase in mitochondrial and cellular reactive oxygen species and 8-hydroxydeoxyguanosine levels, respectively. In the studies distinguishing between oxidative and direct DNA damage, 1 h pretreatment with glutathione (GSH) or the antioxidant Trolox showed a decrease in CEES-induced oxidative stress and oxidative DNA damage. However, only GSH pretreatment decreased CEES-induced total DNA damage measured by comet assay, H2A.X and p53 phosphorylation, and total p53 levels. This was possibly due to the formation of GSH-CEES conjugates detected by LC-MS analysis. Together, our results show that CEES causes both direct and oxidative DNA damage, suggesting that to rescue SM-caused skin injuries, pleiotropic agents (or cocktails) are needed that could target multiple pathways of mustard skin toxicities. (C) 2011 Elsevier Inc. All rights reserved.