Activation of the chemosensing transient receptor potential channel A1 (TRPA1) by alkylating agents
Activation of the chemosensing transient receptor potential channel A1 (TRPA1) by alkylating agents
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烷化剂激活化学传感瞬时受体电位通道 A1 (TRPA1)
DOI：
10.1007/s00204-014-1414-4
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发表时间：
2015
影响因子：
6.1
通讯作者：
Steinritz D
中科院分区：
文献类型：
--
作者：
Stenger B;Zehfuss F;Mückter H;Schmidt A;Balszuweit F;Schäfer E;Büch T;Gudermann T;Thiermann H;Steinritz D
瞬时受体电位锚蛋白1（TRPA 1）阳离子通道在不同组织中表达，包括皮肤、肺和神经组织。最近的报告确定TRPA 1作为一个传感器的有毒物质，暗示在分子毒理学的功能作用。TRPA 1被各种潜在有害的亲电子物质激活。化学战剂硫芥子气（SM）是一种高度反应性的烷化剂，可与许多生物目标结合。虽然SM被称为近200年来，详细的知识造成的暴露的病理生理学是缺乏的。没有特定的治疗方法。在这项研究中，我们研究了烷化剂2-氯乙基乙基硫醚（CEES，SM促进效应的模型物质）和SM是否能够激活TRPA 1通道。CEES诱导TRPA 1表达细胞内钙离子浓度（[Ca 2 +]i）显著增加，但TRPA 1阴性细胞内钙离子浓度（[Ca 2 +]i）无明显变化。TRP通道阻断剂AP 18可减少CEES诱导的钙内流。与野生型细胞相比，永久表达TRPA 1的HEK 293细胞对CEES的细胞毒性作用更敏感。在低CEES浓度下，CEES诱导的细胞毒性被AP 18阻止。使用SM的概念验证实验导致HEK 293-A1-E细胞中[Ca 2 +] i的显著增加。内源性表达TRPA 1的人A549肺上皮细胞在CEES暴露后与瞬时钙内流反应。CEES依赖性钙反应被AP 18减弱。总之，我们的结果表明，烷化剂能够激活TRPA 1。TRPA 1的抑制抵消了细胞毒性，因此可以代表减轻SM诱导的细胞损伤的可行方法。
The transient receptor potential ankyrin 1 (TRPA1) cation channel is expressed in different tissues including skin, lung and neuronal tissue. Recent reports identified TRPA1 as a sensor for noxious substances, implicating a functional role in the molecular toxicology. TRPA1 is activated by various potentially harmful electrophilic substances. The chemical warfare agent sulfur mustard (SM) is a highly reactive alkylating agent that binds to numerous biological targets. Although SM is known for almost 200 years, detailed knowledge about the pathophysiology resulting from exposure is lacking. A specific therapy is not available. In this study, we investigated whether the alkylating agent 2-chloroethyl-ethylsulfide (CEES, a model substance for SM-promoted effects) and SM are able to activate TRPA1 channels. CEES induced a marked increase in the intracellular calcium concentration ([Ca2+]i) in TRPA1-expressing but not in TRPA1-negative cells. The TRP-channel blocker AP18 diminished the CEES-induced calcium influx. HEK293 cells permanently expressing TRPA1 were more sensitive toward cytotoxic effects of CEES compared with wild-type cells. At low CEES concentrations, CEES-induced cytotoxicity was prevented by AP18. Proof-of-concept experiments using SM resulted in a pronounced increase in [Ca2+]iin HEK293-A1-E cells. Human A549 lung epithelial cells, which express TRPA1 endogenously, reacted with a transient calcium influx in response to CEES exposure. The CEES-dependent calcium response was diminished by AP18. In summary, our results demonstrate that alkylating agents are able to activate TRPA1. Inhibition of TRPA1 counteracted cellular toxicity and could thus represent a feasible approach to mitigate SM-induced cell damage.