Role of factor VIII-von Willebrand factor and fibronectin in the interaction of platelets in flowing blood with monomeric and fibrillar human collagen types I and III.
Role of factor VIII-von Willebrand factor and fibronectin in the interaction of platelets in flowing blood with monomeric and fibrillar human collagen types I and III.
复制标题
因子 VIII-von Willebrand 因子和纤连蛋白在流动血液中血小板与单体和纤维状人类 I 型和 III 型胶原蛋白相互作用中的作用。
DOI：
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发表时间：
1985
影响因子：
15.9
通讯作者：
J. Sixma
中科院分区：
文献类型：
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作者：
W. Houdijk;K. Sakariassen;P. Nievelstein;J. Sixma
从人脐动脉中提纯的I型和III型胶原单体与血小板的粘附性在灌注室中进行了研究。为此，玻璃盖片上涂有20-30微克/厘米2的I型和III型胶原蛋白，方法是用润饰气刷喷洒这些胶原蛋白的溶液。随着灌流时间的延长，血小板沉积增加。在前3分钟，两种胶原蛋白的粘附性相似，但在3分钟后，由于血栓的形成，在III型胶原蛋白上出现了更多的血小板沉积。Von Willebrand病患者的血浆或不含纤维粘连蛋白的血浆对S-1切变率为800时的粘附力有强烈的抑制作用。将纯化的纤维结合蛋白加入不含纤维结合蛋白的血浆中，使粘附性恢复到正常血浆的水平。正常血浆中的血小板沉积随着切变率的增加而增加。在490 S-1时，VWD血浆中的血小板沉积正常，但在较高的切变率下不会增加。在研究的490到1,300 S-1的所有切变率下，无纤维连接蛋白血浆中的血小板沉积减少。加入纯化因子VIII-von Willebrand因子复合体(FVIII-VWF)和纤维连接蛋白的人血清白蛋白溶液(HAS)灌流后，可获得与正常血浆相似的血小板沉积。将胶原与FVIII-VWF预先孵育，再用含有纤维连接蛋白的HAS灌流，或者反过来，与纤维连接蛋白预先孵育，再用含有FVIII-VWF的HAS灌流，也能产生类似于正常血浆中观察到的粘连。在与FVIII-VWF和纤维连接蛋白预先孵育，然后单独与HAS灌流后，也观察到类似的粘连。先用FVIII-VWF，然后用纤维连接蛋白，或先用纤维连接蛋白，再用FVIII-VWF，然后用HAS灌流，也能产生与正常血浆相似的粘附性。这些数据表明，血小板与I型和III型单体胶原的粘附性依赖于FVIII-VWF和纤维连接蛋白。FVIII-VWF仅在相对较高的切变率下需要；纤维连接蛋白也在相对较低的切变率下需要。它们在血小板黏附中的互补作用提示FVIII-VWF和纤维连接蛋白在胶原上有不同的结合部位。血小板在I型和III型胶原纤维上的沉积也被研究。初始黏附以表面覆盖率表示，与单体胶原相似，但血栓形成明显增强。在VWD血浆和无纤维连接蛋白血浆中，800℃时S(-1)血浆对胶原纤维的粘附性受到损害，并通过在无纤维连接蛋白血浆中加入纯化的纤维连接蛋白而恢复。当使用HAS进行灌流时，只需添加FVIII-VWF即可获得与纤维胶原的最佳粘附性；添加纤维连接蛋白则不起作用。这些数据与上面描述的单体胶原蛋白的研究形成了对比，在这些研究中，需要同时添加FVIII-VWF和纤维连接蛋白。这些数据也与血浆中FVIII-VWF和纤维连接蛋白都是最佳粘附胶原蛋白的观察结果形成了鲜明对比。
Platelet adhesion to monomeric collagen types I and III, which were purified from human umbilical arteries, was studied in a perfusion chamber under well defined flow conditions. For this purpose, glass coverslips were coated with 20-30 micrograms/cm2 of collagen types I and III by spraying a solution of these collagens with a retouching air brush. Platelet deposition increased with the time of perfusion. Adhesion to both collagen types was similar in the first 3 min, but increased platelet deposition occurred on collagen type III after 3 min due to thrombus formation. Adhesion at a shear rate of 800 s-1 was strongly impaired with plasma of a patient with von Willebrand's disease or with fibronectin-free plasma. Addition of purified fibronectin to fibronectin-free plasma restored adhesion to the level obtained with normal plasma. Platelet deposition in normal plasma increased with increasing shear rates. Platelet deposition in VWD-plasma was normal at 490 s-1, but there was no increase at higher shear rates. Platelet deposition in fibronectin-free plasma was diminished at all shear rates studied from 490 to 1,300 s-1. Perfusion with a human albumin solution (HAS) to which purified Factor VIII-von Willebrand factor complex (FVIII-VWF) and fibronectin had been added gave similar platelet deposition as with normal plasma. Preincubation of collagen with FVIII-VWF and perfusion with HAS containing fibronectin, or, conversely, preincubation with fibronectin and perfusion with HAS containing FVIII-VWF, also resulted in adhesion similar to that observed in normal plasma. Similar adhesion was also observed after preincubation with both FVIII-VWF and fibronectin and subsequent perfusion with HAS alone. Sequential preincubations with first FVIII-VWF and then fibronectin, or with first fibronectin and then FVIII-VWF followed by perfusion with HAS, also gave a similar adhesion as observed with normal plasma. These data indicate that platelet adhesion to monomeric collagen types I and III is dependent on both FVIII-VWF and fibronectin. FVIII-VWF is only required at relatively high shear rates; fibronectin also at relatively low shear rates. Their complementary role in platelet adhesion suggests separate binding sites for FVIII-VWF and fibronectin on collagen. Platelet deposition on performed fibrils of collagen types I and III was also studied. Initial adhesion expressed as percentage surface coverage was similar to that found with monomeric collagen, but thrombus formation was much enhanced. Adhesion on fibrillar collagen at 800 s(-1) was impaired in VWD-plasma and fibronectin-free plasma, and was restored by addition of purified fibronectin to fibronectin-free plasma. When perfusions were performed with HAS, only addition of FVIII-VWF was required for optimal adhesion to fibrillar collagen; addition of fibronectin had no effect. These data are in contrast to the studies with monomeric collagens described above, in which the addition of both FVIII-VWF and fibronectin was required. These data are also in contrast to the observation that in plasma both FVIII-VWF and fibronectin are required for optimal adhesion to fibrillar collagen.