Distinct Ca2+ Requirement for NO Production between Proteinase-Activated Receptor 1 and 4 (PAR1 and PAR4) in Vascular Endothelial Cells
Distinct Ca2+ Requirement for NO Production between Proteinase-Activated Receptor 1 and 4 (PAR1 and PAR4) in Vascular Endothelial Cells
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血管内皮细胞中蛋白酶激活受体 1 和 4（PAR1 和 PAR4）产生 NO 的不同 Ca2 需求
DOI：
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发表时间：
2007
影响因子：
3.5
通讯作者：
H. Kanaide
中科院分区：
文献类型：
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作者：
K. Hirano;N. Nomoto;M. Hirano;Fumi Momota;A. Hanada;H. Kanaide
蛋白酶激活受体1和4（PAR 1和PAR 4）是介导凝血酶诱导内皮细胞产生NO的主要受体。其激活后的细胞内信号传导仍有待阐明。本研究提供了第一个证据，不同的Ca 2+的要求之间的PAR 1和PAR 4的NO生产。在猪主动脉和人脐静脉内皮细胞中，激活肽（PAR 1-AP）激活PAR 1可升高胞浆Ca 2+浓度（[Ca 2 +]i）并激活NO产生，而对牛主动脉内皮细胞影响不大。PAR 4-AP激活PAR 4后，在三种内皮细胞中均能诱导NO产生，但[Ca 2 +]i无明显升高。1，2-二（2-氨基苯氧基）乙烷-N，N，N′，N′-四乙酸（BAPTA）可显著抑制PAR 1介导的NO生成，而PAR 4介导的NO生成则不受BAPTA的影响。百日咳毒素可显著抑制PAR 1和PAR 4激活后NO的产生，但对Gαq/11抑制剂YM 254890 [（1 R）-1-{（3S，6S，9 S，12 S，18 R，21 S，22 R）-21-乙酰氨基-18-苄基-3-[（1 R）-1-甲氧基乙基]-4，9，10，12，16，22-六甲基-15-亚甲基-2，5，8，11，14，17，20-七氧代-1，19-二氧杂-4，7，10，13，16-五氮杂环二十二烷-6-基}-2-甲基丙基rel-（2S，3R）-2-乙酰氨基-3-羟基-4-甲基戊酸酯]。然而，YM 254890消除了PAR 1介导的Ca 2+信号。PAR 4介导的NO产生被磷脂酰肌醇-3激酶（PI 3 K）和Akt的抑制剂以及Akt的显性失活突变体显著抑制。PAR 1介导的NO产生对PI 3 K抑制剂相对耐受。免疫印迹分析显示，在PAR 4刺激后Akt和内皮NO合酶的磷酸化瞬时增加。总之，PAR 1和PAR 4参与不同的信号转导机制来激活血管内皮细胞中NO的产生。PAR 4优选地以主要不依赖于Ca 2+但依赖于PI 3 K/Akt途径的方式激活Gαi/o并诱导NO产生，而PAR 1激活Ca 2+依赖性和非依赖性机制。
Proteinase-activated receptors 1 and 4 (PAR1 and PAR4) are the major receptors mediating thrombin-induced NO production in endothelial cells. The intracellular signaling following their activation still remains to be elucidated. The present study provides the first evidence for the distinct Ca2+ requirement for the NO production between PAR1 and PAR4. The activation of PAR1 by the activating peptide (PAR1-AP) elevated cytosolic Ca2+ concentrations ([Ca2+]i) and activated NO production in porcine aortic and human umbilical vein endothelial cells, whereas it had little effect on bovine aortic endothelial cells. PAR4 activation by PAR4-AP consistently induced NO production without an appreciable [Ca2+]i elevation in three types of endothelial cells. The PAR1-mediated NO production was significantly inhibited by 1,2-bis(2-aminophenoxy)ethane-N,N,N′,N′-tetraacetic acid (BAPTA), whereas the PAR4-mediated NO production was resistant. NO production following the PAR1 and PAR4 activation was significantly inhibited by pertussis toxin, but it was resistant to a Gαq/11 inhibitor, YM254890 [(1R)-1-{(3S,6S,9S,12S,18R,21S,22R)-21-acetamido-18-benzyl-3-[(1R)-1-methoxyethyl]-4,9,10,12,16,22-hexamethyl-15-methylene-2,5,8,11,14,17,20-heptaoxo-1,19-dioxa-4,7,10,13,16-pentaazacyclodocosan-6-yl}-2-methylpropyl rel-(2S,3R)-2-acetamido-3-hydroxy-4-methylpentanoate]. However, YM254890 abrogated the PAR1-mediated Ca2+ signal. PAR4-mediated NO production was substantially inhibited by the inhibitors of phosphotidylinositol-3 kinase (PI3K) and Akt, as well as by the dominant negative mutant of Akt. The PAR1-mediated NO production was relatively resistant to inhibitors of PI3K. An immunoblot analysis revealed a transient increase in the phosphorylation of Akt and endothelial NO synthase following the PAR4 stimulation. In conclusion, PAR1 and PAR4 engage distinct signal transduction mechanisms to activate NO production in vascular endothelial cells. PAR4 preferably activates Gαi/o and induced NO production in a manner mostly independent of Ca2+ but dependent on the PI3K/Akt pathway, whereas PAR1 activates both the Ca2+-dependent and -independent mechanisms.
影响因子：
20.3
作者：
Kataoka, H;Hamilton, JR;Coughlin, SR
通讯作者：
Coughlin, SR