FⅫ在促凝表面的接触活化动力学研究

摘要

Objective In order to explore the microscopic mechanism of pathological clotting plasma interaction,we designed the intrinsic coagulation contact activation experiments to study how the endogenous coagulation factors induced procoagulant surface and the induced conditions.Methods Clean glass beads and silane glass beads were used,then we got different hydrophilic properties of procoagulant particle surface to determine the relationship between the activation of factor Ⅻ and hydrophilic properties of surface;buffer PBS was used to wash procoagulant surface,and then the surface was contact with platelet-poor plasma.We compared the clotting time to study the effects of plasma protein on clotting time.Results (1) Activation of FⅫ-FⅫa in the hydrophilic surface exhibited specific properties,the activation reaction in a hydrophobic surface significantly decreased (P<0.0001),and much more FⅫa was activated when FⅫ was contacted with a hydrophobic surface;(2) The FⅫ activation rate on hydrophobic surface was significantly decayed,and less products were obtained,however,activation FⅫ with a hydrophobic surface was binding tightly (P<0.0001),which was consistent with the result about the protein with a hydrophobic surface adhesion;(3) When the procoagulant activation was on the surface of 500 mm2 and 30 min incubated time,the clotting time and the active product of factor Ⅻ were much more stable.Conclusions Studies had shown that the surface energy depended on "catalytic potential" to promote FⅫ-FⅫa activation;When the coagulation surface area was the same,the activation rate and the amount of activation within PPP plasma FⅫ were significantly lower on the hydrophobic surface than the hydrophilic surface.On the contrary of the protein with a hydrophobic surface activation,FⅫ-full plasma activation on the hydrophobic surface were more slow than the hydrophilic surface which results in the appearance of a surface-energy-dependent catalytic potential.%目的 为了探索病理性凝血血浆间相互作用的微观机制,设计内源性凝血接触激活实验,研究促凝性表面诱发的内源性凝血的条件以及诱发的主要因素.方法 本文采用了干净的玻璃珠和硅烷化玻璃珠,从而得到亲水性能不同的促凝颗粒表面,来确定因子Ⅶ的激活与亲水性的关系;利用缓冲液PBS冲洗促凝剂表面,再与乏血小板血浆反应,对比各凝血时间的变化,研究血浆蛋白对凝血时间的影响.结果 (1) 在亲水表面FⅫ-FⅫa的激活表现出特异性,在疏水表面的活化反应相对明显减弱(P<0.0001),FⅫ与疏水促凝剂接触激活获得更多FⅫa;(2) 在疏水表面FⅫ的激活率和产物都是显著衰减,但是活化FⅫ与疏水表面结合紧密(P<0.0001),与蛋白质和疏水表面黏附更紧密的结论一致;(3) FⅫ与500mm2促凝表面在孵育30min时凝血时间和FⅫ的活化产物更为稳定.结论 促凝剂表面能量依赖 "催化潜力"促使FⅫ-FⅫa的激活;凝血表面积相同的情况下,PPP血浆内FⅫ的活化率及活化量在疏水促凝剂表面明显低于亲水性表面;与蛋白质和疏水表面更易吸附活化相反,在含疏水促凝剂表面的全血浆中FⅫ的活化明显比亲水性的慢,从而产生了一种表面能依赖的催化潜力的外观.