Martinez-Sanchez Julia, Torramade-Moix Sergi, Moreno-Castaño Ana Belén, Llobet Dolors, Jerez-Dolz Didac, Sanchez Pablo, Carrasco Marina, Mojal Sergi, Moret Carla, Camacho Mercedes, Soria José Manuel, Palomo Marta, Martin-Fernandez Laura, Vidal Francisco, Escolar Gines, Diaz-Ricart Maribel, Souto Juan Carlos
Department of Pathology, Hemostasis and Erythropathology Laboratory, Hematopathology, Centre de Diagnòstic Biomèdic (CDB), Hospital Clínic de Barcelona, Institut d'Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS), Universitat de Barcelona, Barcelona, Spain.
Barcelona Endothelium Team, Barcelona, Spain.
TH Open. 2024 Nov 30;8(4):e329-e339. doi: 10.1055/s-0044-1800798. eCollection 2024 Oct.
Association between global platelet function and the risk of venous thromboembolic disease (VTE) has been proposed, though the mechanisms do not involve increased platelet aggregation. However, platelet adhesiveness has not been systematically explored in VTE patients. To evaluate platelet adhesive functions in VTE patients. Platelet adhesion was evaluated by using whole blood samples from VTE patients, selected based on short closure times on the PFA-100 ( = 54), and matched healthy individuals ( = 57) in: (i) the PFA-100, (ii) a cone plate analyzer (CPA), on a plastic surface, (iii) microfluidic devices, with two- and three-dimensional evaluation, and (iv) membrane glycoprotein analysis. Intraplatelet signaling was evaluated in isolated collagen type I (Col-I) activated platelets and platelets adhered on Col-I or von Willebrand factor (VWF) coated coverslips under flow. VWF antigen and ADAMTS-13 activity were measured in plasma samples. PFA-100 closure times remained significantly shorter in patients. The CPA test showed a significant increase in the platelet aggregates size when using blood from VTE patients. Platelet adhesion on Col-I revealed a higher area covered by platelets and increased aggregate volume when exposed to samples from VTE patients. Protein P-ZAP70/SYK72 showed a phosphorylation level significantly increased in patients' platelets. Plasma VWF was significantly elevated in VTE patients. Platelets from VTE patients exhibit a proadhesive phenotype under flow conditions potentially related to the shortened occlusion times with the PFA-100. This enhanced adhesiveness may be explained by higher intraplatelet ZAP70/SYK72 phosphorylation and increased plasma VWF in patients. Therefore, primary hemostasis plays a significant role in the pathophysiology of VTE.
尽管全球血小板功能与静脉血栓栓塞性疾病（VTE）风险之间的关联已被提出，但其机制并不涉及血小板聚集增加。然而，VTE患者的血小板黏附性尚未得到系统研究。
为评估VTE患者的血小板黏附功能。
通过使用VTE患者的全血样本评估血小板黏附，这些患者基于PFA-100上的短封闭时间进行选择（n = 54），并与匹配的健康个体（n = 57）进行比较，具体如下：（i）PFA-100；（ii）在塑料表面的锥板分析仪（CPA）；（iii）具有二维和三维评估的微流控装置；（iv）膜糖蛋白分析。在分离的I型胶原（Col-I）激活的血小板以及在流动条件下黏附于Col-I或血管性血友病因子（VWF）包被盖玻片上的血小板中评估血小板内信号传导。在血浆样本中测量VWF抗原和ADAMTS-13活性。
患者的PFA-100封闭时间仍然显著缩短。CPA测试显示，使用VTE患者的血液时，血小板聚集体大小显著增加。当暴露于VTE患者的样本时，Col-I上的血小板黏附显示血小板覆盖的面积更大且聚集体体积增加。蛋白P-ZAP70/SYK72在患者血小板中的磷酸化水平显著升高。VTE患者的血浆VWF显著升高。
VTE患者的血小板在流动条件下表现出促黏附表型，这可能与PFA-100的缩短闭塞时间有关。这种增强的黏附性可能是由于患者血小板内ZAP70/SYK72磷酸化水平较高以及血浆VWF增加所致。因此，初级止血在VTE的病理生理学中起重要作用。
