Department of Biochemistry, Maastricht University MedicalCentre, Maastricht, The Netherlands.
Blood. 2011 Jan 13;117(2):651-60. doi: 10.1182/blood-2010-01-262683. Epub 2010 Oct 29.
A microscopic method was developed to study the role of platelets in fibrin formation. Perfusion of adhered platelets with plasma under coagulating conditions at a low shear rate (250(-1)) resulted in the assembly of a star-like fibrin network at the platelet surface. The focal fibrin formation on platelets was preceded by rises in cytosolic Ca(2+), morphologic changes, and phosphatidylserine exposure. Fibrin formation was slightly affected by α(IIb)β(3) blockage, but it was greatly delayed and reduced by the following: inhibition of thrombin or platelet activation; interference in the binding of von Willebrand factor (VWF) to glycoprotein Ib/V/IX (GpIb-V-IX); plasma or blood from patients with type 1 von Willebrand disease; and plasma from mice deficient in VWF or the extracellular domain of GpIbα. In this process, the GpIb-binding A1 domain of VWF was similarly effective as full-length VWF. Prestimulation of platelets enhanced the formation of fibrin, which was abrogated by blockage of phosphatidylserine. Together, these results show that, in the presence of thrombin and low shear flow, VWF-induced activation of GpIb-V-IX triggers platelet procoagulant activity and anchorage of a star-like fibrin network. This process can be relevant in hemostasis and the manifestation of von Willebrand disease.
一种研究血小板在纤维蛋白形成中作用的显微镜方法被开发出来。在低切变率(250(-1))下,将附着的血小板用血浆灌注,并在凝固条件下进行,结果在血小板表面形成了星形纤维蛋白网络。血小板表面的局灶性纤维蛋白形成之前,细胞内钙离子浓度升高、形态发生变化和磷脂酰丝氨酸暴露。α(IIb)β(3)阻断对纤维蛋白形成略有影响,但被以下因素大大延迟和减少:抑制凝血酶或血小板激活;干扰血管性血友病因子(VWF)与糖蛋白 Ib/V/IX(GpIb-V-IX)的结合;1 型血管性血友病患者的血浆或血液;以及缺乏 VWF 或 GpIbα细胞外结构域的小鼠血浆。在此过程中,VWF 的 GpIb 结合 A1 结构域与全长 VWF 同样有效。血小板的预刺激增强了纤维蛋白的形成,而血小板磷脂酰丝氨酸的阻断则削弱了纤维蛋白的形成。综上所述,这些结果表明,在凝血酶和低切变流的存在下,VWF 诱导的 GpIb-V-IX 激活触发血小板促凝活性和星形纤维蛋白网络的锚定。这个过程在止血和血管性血友病的表现中可能是相关的。