Mendelsohn M E, O'Neill S, George D, Loscalzo J
Cardiovascular Division, Brigham and Women's Hospital, Boston, Massachusetts 02115.
J Biol Chem. 1990 Nov 5;265(31):19028-34.
Because of the central role of fibrinogen binding in platelet aggregation and recent evidence implicating S-nitrosothiol compounds in the platelet inhibitory effects of endogenous and exogenous organic nitrate compounds, we examined the effect of the S-nitrosothiol S-nitroso-N-acetylcysteine (SNOAC) on fibrinogen binding to gel-filtered human platelets. We found that SNOAC markedly inhibited the binding of fibrinogen to normal human platelets in a dose-dependent fashion and that this inhibitory effect was the result of both an increase in the apparent Kd of the platelet receptor for the fibrinogen molecule (from 6.8 x 10(-7) to 1.8 x 10(-6) M, a 2.7-fold increase) and a decrease in the total number of fibrinogen molecules bound to the platelet (from 76,200 to 38,250, a 50% decrease). In addition, we noted a rapid, dose-dependent rise in platelet cyclic GMP levels following exposure of platelets to SNOAC which was significantly inversely correlated with fibrinogen binding and was accompanied by inhibition of intracellular calcium flux in response to a variety of platelet agonists. Similar dose-dependent inhibition of fibrinogen binding was found in the presence of cyclic GMP analogues and was significantly enhanced by inhibition of platelet cyclic GMP phosphodiesterase. These results describe the inhibition of platelet fibrinogen binding by an S-nitrosothiol compound, help define the biochemical mechanism by which S-nitrosothiols inhibit platelet aggregation, and lend support to the view that cyclic GMP is an important inhibitory intracellular mediator in human platelets.
由于纤维蛋白原结合在血小板聚集中的核心作用,以及最近有证据表明S-亚硝基硫醇化合物在内源性和外源性有机硝酸盐化合物的血小板抑制作用中起作用,我们研究了S-亚硝基硫醇S-亚硝基-N-乙酰半胱氨酸(SNOAC)对纤维蛋白原与凝胶过滤的人血小板结合的影响。我们发现SNOAC以剂量依赖性方式显著抑制纤维蛋白原与正常人血小板的结合,并且这种抑制作用是纤维蛋白原分子的血小板受体表观解离常数增加(从6.8×10^(-7)至1.8×10^(-6) M,增加2.7倍)以及与血小板结合的纤维蛋白原分子总数减少(从76,200至38,250,减少50%)的结果。此外,我们注意到血小板暴露于SNOAC后,血小板环鸟苷酸(cGMP)水平迅速、剂量依赖性升高,这与纤维蛋白原结合显著负相关,并伴随着对多种血小板激动剂的细胞内钙通量抑制。在环鸟苷酸类似物存在的情况下发现了类似的剂量依赖性纤维蛋白原结合抑制,并且通过抑制血小板环鸟苷酸磷酸二酯酶显著增强。这些结果描述了一种S-亚硝基硫醇化合物对血小板纤维蛋白原结合的抑制作用,有助于确定S-亚硝基硫醇抑制血小板聚集的生化机制,并支持环鸟苷酸是人类血小板中一种重要的抑制性细胞内介质的观点。