Involvement of STIM1 in the Proteinase-Activated Receptor 1-Mediated Ca2+ Influx in Vascular Endothelial Cells
Involvement of STIM1 in the Proteinase-Activated Receptor 1-Mediated Ca2+ Influx in Vascular Endothelial Cells
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DOI：
10.1002/jcb.22279
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发表时间：
2009-10-01
影响因子：
4
通讯作者：
Hanada, Akiko
中科院分区：
文献类型：
--
作者：
Hirano, Katsuya;Hirano, Mayumi;Hanada, Akiko
凝血酶通过激活血管内皮细胞中的蛋白酶激活受体1（PAR（1））增加胞浆Ca 2+浓度并诱导NO产生。钙池操纵的钙内流是包括内皮细胞在内的非兴奋性细胞中的主要钙内流途径，并且已报道其在凝血酶诱导的内皮细胞中的钙信号转导中起作用。最近的研究已经确定基质相互作用分子1（STIM 1）作为一个传感器的商店网站钙含量，从而调节商店操作的钙内流。然而，STIM 1在凝血酶诱导的内皮细胞Ca 2+内流和NO产生中的功能作用仍有待阐明。Fura-2和diaminorhodamine-4 M荧光法被用来评估凝血酶诱导的变化，细胞内Ca 2+浓度和NO的生产，分别在猪主动脉内皮细胞转染小干扰RNA（siRNA）靶向STIM 1。靶向STIM 1的siRNA抑制STIM 1的表达和毒胡萝卜素诱导的Ca 2+内流。STIM 1表达的抑制程度与Ca 2+内流的抑制程度相关。STIM 1的敲低与凝血酶诱导的Ca 2+内流的实质性抑制和NO产生的部分减少有关。凝血酶诱导的Ca 2+内流对Ca 2+内流抑制剂表现出与毒胡萝卜素诱导的Ca 2+内流相似的敏感性。本研究首次证明STIM 1在PAR（1）介导的内皮细胞Ca ~（2+）内流和Ca ~（2+）依赖的NO生成中起关键作用。J.细胞。108：499-507，2009. (C)2009年威利-利斯轮胎
Thrombin increases the cytosolic Ca2+ concentrations and induces NO production by activating proteinase-activated receptor 1 (PAR(1)) in vascular endothelial cells. The store-operated Ca2+ influx is a major Ca2+ influx pathway in non-excitable cells including endothelial cells and it has been reported to play a role in the thrombin-induced Ca2+ signaling in endothelial cells. Recent studies have identified stromal interaction molecule 1 (STIM1) to function as a sensor of the store site Ca2+ content, thereby regulating the store-operated Ca2+ influx. However, the functional role of STIM1 in the thrombin-induced Ca2+ influx and NO production in endothelial cells still remains to be elucidated. Fura-2 and diaminorhodamine-4M fluorometry was utilized to evaluate the thrombin-induced changes in cytosolic Ca2+ concentrations and NO production, respectively, in porcine aortic endothelial cells transfected with small interfering RNA (siRNA) targeted to STIM1. STIM1-targeted siRNA suppressed the STIM1 expression and the thapsigargin-induced Ca2+ influx. The degree of suppression of the STIM1 expression correlated well to the degree of suppression of the Ca2+, influx. The knockdown of STIM1 was associated with a substantial inhibition of the Ca2+ influx and a partial reduction of the NO production induced by thrombin. The thrombin-induced Ca2+, influx exhibited the similar sensitivity toward the Ca2+ influx inhibitors to that seen with the thapsigargin-induced Ca2+ influx. The present study provides the first evidence that STIM1 plays a critical role in the PAR(1)-mediated Ca2+ influx and Ca2+ dependent component of the NO production in endothelial cells. J. Cell. Biochem. 108: 499-507, 2009. (C) 2009 Wiley-Liss, tire.