Department of Internal Medicine, The Vrinnevi Hospital, Norrköping, Sweden.
Department of Internal Medicine, The Vrinnevi Hospital, Norrköping, Sweden.
Thromb Res. 2020 Nov;195:1-7. doi: 10.1016/j.thromres.2020.06.033. Epub 2020 Jun 23.
Strong agonist provocation in vitro creates small procoagulant platelets characterized by down-regulated fibrinogen receptors as judged from surface αβ activation specific antibody (PAC-1). They further show increased surface Annexin V (binds to platelet membrane phosphatidylserine), lysosomal-associated membrane protein 1 (LAMP-1) (indicates lysosomal release) and exhibit disturbed mitochondria integrity as estimated from mitochondrial transmembrane potential changes. We postulated that some circulating platelets activate continuously thereby forming procoagulant populations in vivo. This study aimed to identify such platelets in diabetes type 2 a condition predisposing for thrombotic events.
A linear Percoll™ gradient covering the density span 1.090 to 1.040 kg/L was used to separate whole blood platelets from type 2 diabetic subjects (n = 12) into 17 density subpopulations. The gradient contained theophylline, prostaglandin E and EDTA to prevent platelet activation in vitro. A multi-colour flow cytometer was employed for analysing the characteristics mentioned above for all density separated small-sized platelet subfractions.
Small platelets were enriched in medium-dense subfractions (nos. 10-13) (1.065-1.053 kg/L). Their PAC-1 activities were significant lower (p < 0.001) as compared to other small-sized subpopulations. They further exposed enhanced surface Annexin V and LAMP-1 together with lower mitochondrial transmembrane potentials. In diabetes type 2 such small circulating platelets showed procoagulant features.
体外强激动剂刺激可产生小促凝血小板,这些血小板的特征是表面纤维蛋白原受体下调,可根据表面αβ激活特异性抗体(PAC-1)判断。它们还表现出表面 Annexin V(与血小板膜磷脂酰丝氨酸结合)和溶酶体相关膜蛋白 1(LAMP-1)增加(表示溶酶体释放),并且根据线粒体跨膜电位变化估计线粒体完整性受损。我们假设一些循环血小板持续激活,从而在体内形成促凝群体。本研究旨在鉴定 2 型糖尿病患者(易发生血栓事件的一种情况)体内的这种血小板。
使用线性 Percoll™梯度(密度范围为 1.090 至 1.040 kg/L)将来自 12 例 2 型糖尿病患者的全血血小板分离成 17 个密度亚群。梯度中含有茶碱、前列腺素 E 和 EDTA,以防止体外血小板激活。采用多色流式细胞仪分析所有密度分离的小血小板亚群的上述特征。
小血小板在中密亚群(10-13 号)(1.065-1.053 kg/L)中富集。与其他小尺寸亚群相比,它们的 PAC-1 活性显著降低(p < 0.001)。它们进一步显示出增强的表面 Annexin V 和 LAMP-1,以及更低的线粒体跨膜电位。在 2 型糖尿病中,这些小循环血小板表现出促凝特征。