Department of Pharmaceutical Sciences, University of Colorado Denver School of Pharmacy, Aurora, Colorado 80045, USA.
J Pharmacol Exp Ther. 2011 Feb;336(2):450-9. doi: 10.1124/jpet.110.173708. Epub 2010 Oct 25.
Exposure to chemical warfare agent sulfur mustard (HD) is reported to cause GSH depletion, which plays an important role in HD-linked oxidative stress and skin injury. Using the HD analog 2-chloroethyl ethyl sulfide (CEES), we evaluated the role of GSH and its efficacy in ameliorating CEES-caused skin injury. Using mouse JB6 and human HaCaT epidermal keratinocytes, we observed both protective and therapeutic effects of exogenous GSH (1 or 10 mM) in attenuating a CEES-caused decrease in cell viability and DNA synthesis, as well as S and G(2)M phase arrest in cell cycle progression. However, the protective effect of GSH was stronger than its ability to reverse CEES-induced cytotoxic effect. The observed effect of GSH could be associated with an increase in intracellular GSH levels after its treatment before or after CEES exposure, which strongly depleted cellular GSH levels. N-Acetyl cysteine, a GSH precursor, also showed both protective and therapeutic effects against CEES-caused cytotoxicity. Buthionine sulfoximine, which reduces cellular GSH levels, caused an increased CEES cytotoxicity in both JB6 and HaCaT cells. In further studies translating GSH effects in cell culture, pretreatment of mice with 300 mg/kg GSH via oral gavage 1 h before topical application of CEES resulted in significant protection against CEES-caused increase in skin bifold and epidermal thickness, apoptotic cell death, and myeloperoxidase activity, which could be associated with increased skin GSH levels. Together, these results highlight GSH efficacy in ameliorating CEES-caused skin injury and further support the need for effective antioxidant countermeasures against skin injury by HD exposure.
据报道，接触化学战剂硫芥（HD）会导致 GSH 耗竭，这在 HD 相关的氧化应激和皮肤损伤中起着重要作用。我们使用 HD 类似物 2-氯乙基乙基硫醚（CEES），评估 GSH 的作用及其在减轻 CEES 引起的皮肤损伤中的功效。使用小鼠 JB6 和人 HaCaT 表皮角质形成细胞，我们观察到外源性 GSH（1 或 10mM）在减轻 CEES 引起的细胞活力和 DNA 合成下降以及 S 和 G2M 期细胞周期进展中具有保护和治疗作用。然而，GSH 的保护作用强于其逆转 CEES 诱导的细胞毒性作用的能力。观察到的 GSH 作用可能与 CEES 暴露前后处理后细胞内 GSH 水平增加有关，这强烈耗尽了细胞内的 GSH 水平。GSH 的前体 N-乙酰半胱氨酸也显示出对 CEES 引起的细胞毒性的保护和治疗作用。还原型谷胱甘肽水平的降低，导致 JB6 和 HaCaT 细胞中 CEES 的细胞毒性增加。在进一步的细胞培养中研究 GSH 作用的研究中，通过口服灌胃在 CEES 局部应用前 1 小时给予小鼠 300mg/kg GSH，可显著防止 CEES 引起的皮肤折叠和表皮厚度增加、细胞凋亡和髓过氧化物酶活性增加，这可能与皮肤 GSH 水平增加有关。总之，这些结果突出了 GSH 在减轻 CEES 引起的皮肤损伤中的功效，并进一步支持了针对 HD 暴露引起的皮肤损伤采取有效抗氧化对策的必要性。
