Proteomic Signature of Thrombin-Activated Platelets After In Vivo Nitric Oxide-Donor Treatment Coordinated Inhibition of Signaling (Phosphatidylinositol 3-Kinase-γ, 14-3-3ζ, and Growth Factor Receptor-Bound Protein 2) and Cytoskeleton Protein Translocation
Proteomic Signature of Thrombin-Activated Platelets After In Vivo Nitric Oxide-Donor Treatment Coordinated Inhibition of Signaling (Phosphatidylinositol 3-Kinase-γ, 14-3-3ζ, and Growth Factor Receptor-Bound Protein 2) and Cytoskeleton Protein Translocation
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DOI：
10.1161/atvbaha.111.231852
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发表时间：
2011-11-01
影响因子：
8.7
通讯作者：
Badimon, Lina
中科院分区：
文献类型：
--
作者：
Pena, Esther;Padro, Teresa;Badimon, Lina
目的——对新一氧化氮（NO）供体的抗血小板特性的深入了解扩大了它们在心血管疾病中的潜在用途。因此，我们报道，在体内实验模型中，口服新的外源性 NO 供体 (LA419) 可显着抑制受损血管壁上的血小板沉积，而不会引起低血压。凝血酶是受损血管上血小板沉积和血栓形成的主要触发因素之一；然而，NO 对凝血酶诱导的血小板活化的影响尚不完全清楚。在此，我们的目的是研究外源NO给药对凝血酶诱导的血小板蛋白主要变化的抑制作用。方法和结果-从一组口服LA419（0.9 mg kg(-1)）或安慰剂8天的猪中获得血小板。将洗涤的血小板与凝血酶 (0.4 NIH U/mL) 一起孵育。然后根据溶解度差异依次提取血小板蛋白，并通过二维电泳、质谱（基质辅助激光解吸电离/飞行时间）、蛋白质印迹和共聚焦免疫荧光进行研究。 NO 处理消除了凝血酶对参与肌动蛋白组装、信号传导和代谢活动的 24 种蛋白质的影响。 NO 治疗可防止凝血酶诱导的凝溶胶蛋白、细丝蛋白、14-3-3 zeta、磷脂酰肌醇 3-激酶-γ 亚型和生长因子受体结合蛋白 2 (Grb2) 的易位。结论 - 我们的结果表明，外源性 NO 供体治疗使血小板对凝血酶激活不太敏感，并通过干扰血小板形状变化机制来抑制血栓形成。 （动脉硬化血栓 Vasc Biol。2011；31：2560-2569。）
Objective-Growing insight into the antiplatelet properties of new nitric oxide (NO) donors has expanded their potential use in cardiovascular diseases. As such, we reported that oral administration of a new exogenous NO donor (LA419) induced significant inhibition of platelet deposition on damaged vascular wall without provoking hypotension in an in vivo experimental model. Thrombin is one of the major triggers of platelet deposition and thrombosis on injured vessels; however, the effects of NO on thrombin-induced platelet activation are not fully known. Here, our aim was to investigate the inhibitory effects of exogenous NO administration on the major changes in platelet proteins induced by thrombin.Methods and Results-Platelets were obtained from a group of swine orally treated with LA419 (0.9 mg kg(-1)) or placebo for 8 days. Washed platelets were incubated with thrombin (0.4 NIH U/mL). Platelet proteins were then sequentially extracted based on differential solubility and studied by two-dimensional electrophoresis, mass spectrometry (matrix-assisted laser desorption ionization/time of flight), Western blot, and confocal immunofluorescence. NO treatment abrogated thrombin effects on 24 proteins involved in actin assembly, signaling, and metabolic activity. NO treatment prevented thrombin-induced translocation of gelsolin, filamin, 14-3-3 zeta, phosphatidylinositol 3-kinase-gamma isoform, and growth factor receptor-bound protein 2 (Grb2).Conclusion-Our results show that exogenous NO donor treatment renders platelets less sensitive to thrombin activation and inhibits thrombosis by interfering with the platelet shape change machinery. (Arterioscler Thromb Vasc Biol. 2011;31:2560-2569.)