TCDD and omeprazole prime platelets through the aryl hydrocarbon receptor (AhR) non-genomic pathway
TCDD and omeprazole prime platelets through the aryl hydrocarbon receptor (AhR) non-genomic pathway
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DOI：
10.1016/j.toxlet.2015.03.005
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发表时间：
2015-05-19
影响因子：
3.5
通讯作者：
Tablin, Fern
中科院分区：
文献类型：
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作者：
Pombo, Monica;Lame, Michael W.;Tablin, Fern
芳烃受体（AhR）在止血中的作用最近受到越来越多的关注。在这里，我们通过 qRT-PCR 和蛋白质印迹证明，人血小板表达 AhR mRNA 和 AhR 蛋白。当与 2,3,7,8-四氯二苯并-对二恶英 (TCDD) 或奥美拉唑一起孵育时，AhR 蛋白水平以剂量依赖性方式增加。在 AhR 激活剂存在的情况下，用嘌呤霉素处理血小板会增加 AhR 蛋白的合成。此外，用任一激活剂处理血小板都会导致血小板激活途径中的两个关键信号分子 p38MAPK 和 cPLA(2) 磷酸化。使用 AhR 竞争性抑制剂 α 萘黄酮和 CH-223191，我们发现 p38MAPK 的磷酸化是 AhR 依赖性的。此外，抑制 p38MAPK 可阻断 TCDD 或奥美拉唑诱导的下游 cPLA(2) 磷酸化。 AhR 激活剂治疗会导致血小板启动，如血小板聚集增加所证明的那样，而血小板聚集可被 AhR 拮抗剂抑制。我们的数据支持血小板 AhR 非基因组途径模型，其中使用 AhR 激活剂治疗会导致 AhR 表达增加、p38MAPK 和 cPLA(2) 磷酸化，从而导致血小板对激动剂产生反应。 (C) 2015 Elsevier Ireland Ltd. 保留所有权利。
The role of the aryl hydrocarbon receptor (AhR) in hemostasis has recently gained increased attention. Here, we demonstrate, by qRT-PCR and western blot, that human platelets express both AhR mRNA and AhR protein. AhR protein levels increase in a dose dependent manner when incubated with either 2,3,7,8-tetrachlorodibenzo- p-dioxin (TCDD) or omeprazole. Treatment of platelets with puromycin blocks increased AhR protein synthesis in the presence of AhR activators. Additionally, treatment of platelets with either activator results in phosphorylation of p38MAPK and cPLA(2), two key signaling molecules in platelet activation pathways. Using the AhR competitive inhibitors alpha naphthoflavone and CH-223191, we show that phosphorylation of p38MAPK is AhR dependent. Further, inhibition of p38MAPK blocks downstream cPLA(2) phosphorylation induced by TCDD or omeprazole. Treatment with AhR activators results in platelet priming, as demonstrated by increased platelet aggregation, which is inhibited by AhR antagonists. Our data support a model of the platelet AhR non-genomic pathway in which treatment with AhR activators results in increased expression of the AhR, phosphorylation of p38MAPK and cPLA(2), leading to platelet priming in response to agonist. (C) 2015 Elsevier Ireland Ltd. All rights reserved.