Thromboxane prostanoid receptor activation impairs endothelial nitric oxide-dependent vasorelaxations: The role of Rho kinase
Thromboxane prostanoid receptor activation impairs endothelial nitric oxide-dependent vasorelaxations: The role of Rho kinase
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DOI：
10.1016/j.bcp.2009.04.022
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发表时间：
2009-08-15
影响因子：
5.8
通讯作者：
Huang, Yu
中科院分区：
文献类型：
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作者：
Liu, Cui Qing;Leung, Fung Ping;Huang, Yu
血栓素前列腺素 (TP) 受体的激活会导致强效血管收缩，从而导致血管张力和血压升高。本研究检验了刺激 TP 受体通过 Rho 激酶依赖性机制损害内皮一氧化氮介导的血管舒张作用的假设。分离 Sprague-Dawley 大鼠的颈总动脉并悬浮在肌动描记器中以测量等长张力的变化。采用成像技术测定原代培养的主动脉内皮细胞中一氧化氮的产生，并通过蛋白质印迹分析测定内皮NOS的磷酸化水平。 9,11-二脱氧-11 α,9 α-环氧甲前列腺素 F-2 α。 (U46619) 抑制异丙肾上腺素诱导的有内皮或无内皮环的松弛。用Rho激酶抑制剂Y27632（2μM）或HA 1077（10μM）治疗仅在有内皮的环中阻止U46619的作用，而蛋白激酶C抑制剂则没有作用。 Rho 激酶抑制剂不影响用 L-NAME 或 1H-[1,2,4]oxadizolo[4,3-alquinoxalin-1-one (ODQ) 处理的完整内皮环中异丙肾上腺素诱导的松弛。异丙肾上腺素刺激培养的大鼠内皮细胞中一氧化氮 (NO) 的产生增加。 U46619 (100 nM) 抑制 NO 产生的增加，并且用 Y27632 处理可阻止这种效应，但不受细胞外钙离子缺失的影响。 U46619 减弱异丙肾上腺素刺激的 eNOS 磷酸化，而 eNOS 对 Y27632 和 HA 1077 的抑制敏感。U46619 介导的作用被 TP 受体拮抗剂 S18886 消除，并且 TP 受体存在于内皮细胞中。目前的结果表明，Rho 激酶激活可能是 U46619 刺激的 TP 受体介导的内皮 NO 产生抑制以及随后的内皮依赖性异丙肾上腺素松弛的主要机制。 (C) 2009 Elsevier Inc. 保留所有权利。
Activation of thromboxane prostanoid (TP) receptors causes potent vasoconstriction, which contributes to increased vascular tone and blood pressure. The present study examined the hypothesis that stimulation of TP receptor impaired endothelial nitric oxide-mediated vasorelaxation via it Rho kinase-dependent mechanism. The common carotid arteries of Sprague-Dawley rats were isolated and suspended in myograph for measurement of changes in isometric tension. The production of nitric oxide in primary cultured aortic endothelial cells was assayed with an imaging technique and phosphorylated levels of endothelial NOS were determined by Western blot analysis. 9,11-dideoxy-11 alpha,9 alpha-epoxyrmethanoprostaglandin F-2 alpha. (U46619) inhibited isoprenaline-induced relaxations in rings with or without endothelium. Treatment with Rho kinase inhibitors, Y27632 (2 mu M) or HA 1077 (10 mu M) prevented the effect of U46619 only in rings with endothelium while protein kinase C inhibitors were without effect. Rho kinase inhibitors did not affect isoprenaline-induced relaxations in endothelium-intact rings treated with L-NAME or 1H-[1,2,4]oxadizolo[4,3-alquinoxalin-1-one (ODQ). Isoprenaline stimulated rises in nitric oxide (NO) production in cultured rat endothelial cells. The increased NO production was inhibited by U46619 (100 nM) and this effect was prevented by treatment with Y27632 but unaffected by the absence of extracellular calcium ions. U46619 attenuated isoprenaline-stimulated phosphorylation of eNOS, which was sensitive to inhibition by Y27632 and HA 1077. U46619-mediated effects were abolished by TP receptor antagonist, S18886 and the TP receptor was present in endothelial cells. The present results demonstrate that Rho kinase activation is likely to be the primary mechanism that underlies the U46619-stimulated TP-receptor-mediated inhibition of endothelial NO production and subsequent endothelium-dependent relaxations to isoprenaline. (C) 2009 Elsevier Inc. All rights reserved.