Xia Z, Frojmovic M M
Department of Physiology, McGill University, Montreal, Quebec, Canada.
Biophys J. 1994 Jun;66(6):2190-201. doi: 10.1016/S0006-3495(94)81015-X.
Shear rate can affect protein adsorption and platelet aggregation by regulating both the collision frequency and the capture efficiency (alpha). These effects were evaluated in well defined shear field in a micro-couette for shear rate G = 10 - 1000 s-1. The rate of protein binding was independent of G, shown for adsorption of albumin to latex beads and PAC1 to activated platelets. The initial aggregation rate for ADP-activated platelets in citrated platelet-rich plasma followed second order kinetics at the initial platelet concentrations between 20,000 and 60,000/microliters. alpha values, which dropped nearly fivefold for a 10-fold increase in G, were approximately proportional to G-1, contrary to a minor drop predicted by the theory that includes protein cross-bridging. Varying ADP concentration did not change alpha of maximally activated platelet subpopulations, suggesting that aggregation between unactivated and activated platelets is negligible. Directly blocking the unoccupied but activated GPIIb-IIIa receptors without affecting pre-bound Fg on "RGD"-activated, fixed platelets (AFP) by GRGDSP or Ro 43-5054 eliminated aggregation, suggesting that cross-bridging of GPIIb-IIIa on adjacent platelets by fibrinogen mediates aggregation. Alpha for AFP remained maximal (approximately 0.24) over 25-75% Fg occupancy, otherwise decreasing rapidly, with a half-maximum occurring at around 2% occupancy, suggesting that very few bound Fg were required to cause significant aggregation.
剪切速率可通过调节碰撞频率和捕获效率(α)来影响蛋白质吸附和血小板聚集。在微库埃特中定义明确的剪切场中,针对剪切速率G = 10 - 1000 s-1评估了这些影响。蛋白质结合速率与G无关,白蛋白吸附至乳胶珠以及PAC1吸附至活化血小板的情况均表明了这一点。在柠檬酸盐富血小板血浆中,ADP活化血小板的初始聚集速率在初始血小板浓度为20,000至60,000/微升时遵循二级动力学。α值在G增加10倍时下降近五倍,与包含蛋白质交联的理论预测的轻微下降相反,α值大致与G-1成正比。改变ADP浓度并未改变最大活化血小板亚群的α值,这表明未活化和活化血小板之间的聚集可忽略不计。通过GRGDSP或Ro 43-5054直接阻断未占据但活化的GPIIb-IIIa受体而不影响“RGD”活化的固定血小板(AFP)上预先结合的Fg,消除了聚集,这表明纤维蛋白原在相邻血小板上对GPIIb-IIIa的交联介导了聚集。对于AFP,在25 - 75%的Fg占据率范围内,α保持最大值(约0.24),否则迅速下降,在约2%的占据率时出现半最大值,这表明只需极少数结合的Fg就能引起显著聚集。