Prothrombin activation in blood coagulation: the erythrocyte contribution to thrombin generation
Prothrombin activation in blood coagulation: the erythrocyte contribution to thrombin generation
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DOI：
10.1182/blood-2012-05-427856
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发表时间：
2012-11-01
期刊：
影响因子：
20.3
通讯作者：
Mann, Kenneth G.
中科院分区：
文献类型：
--
作者：
Whelihan, Matthew F.;Zachary, Vicentios;Mann, Kenneth G.
凝血酶原激活可以通过中间产物甲硫咪唑凝血酶或凝血酶原-2进行。为了评估这两种中间体在组织因子(TF)激活的血液中对凝血酶原激活的贡献，开发了检测甲硫咪唑凝血酶抗凝血酶(MTAT)和α-凝血酶抗凝血酶(αTAT)复合体的免疫分析方法。我们确定TF激活的血液同时产生α-TAT和mTAT。MTat的存在表明，非血小板表面对凝血酶原激活的贡献约为35%。玉米胰酶抑制剂处理的血液被分级，以产生红细胞(RBCs)、富血小板血浆(PRP)、贫血小板血浆(PPP)和黄褐色涂层。与血液相比，用PPP重组的PRP达到生理血小板浓度时，起始期延长了2倍，αTAT的形成速度和程度明显降低。只有在PRP中加入红细胞才能使α-TAT生成正常化。血糖素A阳性细胞的流式细胞术显示，大约0.6%的红细胞群体表达磷脂酰丝氨酸并结合凝血酶原酶(FITC Xa)。这些数据表明，红细胞参与了TF激活的血液中凝血酶的生成，产生了一层膜，通过甲硫唑凝血酶途径支持凝血酶原的激活。(血。2012；120(18)：3837-3845)
Prothrombin activation can proceed through the intermediates meizothrombin or prethrombin-2. To assess the contributions that these 2 intermediates make to prothrombin activation in tissue factor (Tf)-activated blood, immunoassays were developed that measure the meizothrombin antithrombin (mTAT) and alpha-thrombin antithrombin (alpha TAT) complexes. We determined that Tf-activated blood produced both alpha TAT and mTAT. The presence of mTAT suggested that nonplatelet sur-faces were contributing to approximately 35% of prothrombin activation. Corn trypsin inhibitor-treated blood was fractionated to yield red blood cells (RBCs), platelet-rich plasma (PRP), platelet-poor plasma (PPP), and buffy coat. Compared with blood, PRP reconstituted with PPP to a physiologic platelet concentration showed a 2-fold prolongation in the initiation phase and a marked decrease in the rate and extent of alpha TAT formation. Only the addition of RBCs to PRP was capable of normalizing alpha TAT generation. FACS on glycophorin A-positive cells showed that approximately 0.6% of the RBC population expresses phosphatidylserine and binds prothrombinase (FITC Xa.factor Va). These data indicate that RBCs participate in thrombin generation in Tf-activated blood, producing a membrane that supports prothrombin activation through the meizothrombin pathway. (Blood. 2012;120(18):3837-3845)