Normandie Univ, UNIROUEN, INSERM U1096, Rouen University Hospital, Vascular Hemostasis Unit, F 76000 Rouen, France.
Normandie Univ, UNIROUEN, INSERM U1096, Rouen University Hospital, Department of Internal Medicine, Vascular and Thrombosis Unit, F 76000 Rouen, France.
Thromb Res. 2018 Sep;169:87-92. doi: 10.1016/j.thromres.2018.07.021. Epub 2018 Jul 17.
Circulating antiphospholipid antibodies (APL) induce vascular injury and endothelial dysfunction, which are associated with thrombotic events and/or fetal loss. We developed a model in which calibrated automated thrombin generation (CAT) is carried out in wells lined with cultured endothelial cells. Then we investigated how far b2GP1 antibodies provoked thrombin generation (TG) enhancing effects in these cells and/or in blood platelets.
Thrombin generation induced by different concentrations of tissue factor and different levels of endothelial aortic cell confluence was investigated by calibrated automated thrombogram. Endothelial cells were incubated with the purified anti-β2glycoprotein I antibodies of patients with antiphospholipid syndrome (APS). Platelet free plasma and platelet rich plasma were used to study thrombin generation in endothelial cells and platelet reactivity, respectively.
Endothelial cell confluence was negatively correlated with thrombin generation which was dependent on the concentration of APL incubated. Activation of endothelial cells with APL significantly increased thrombin generation triggered by PFP. Triggering by PRP increased thrombinogram parameters. Moreover, anti-β2glycoprotein I antibodies incubated with platelet significantly amplified thrombin formation in PRP and induced platelet activation without tissue factor.
In this in vitro study, we demonstrate the feasibility of using thrombin generation test in cultured endothelial cells and suggest the need to realize adjustments to standardize results. The mechanism of prothrombotic states in APS requires endothelial dysfunction and platelet activation. The quantification of thrombin formation shows that APL incubation induces endothelial injury in cultured cells amplified by platelets.
循环抗磷脂抗体（APL）可诱导血管损伤和内皮功能障碍，与血栓事件和/或胎儿丢失有关。我们开发了一种模型，其中在培养的内皮细胞衬里的孔中进行校准的自动凝血酶生成（CAT）。然后，我们研究了 b2GP1 抗体在这些细胞中和/或在血小板中引起凝血酶生成（TG）增强作用的程度。
通过校准的自动血栓图研究了不同浓度的组织因子和不同水平的内皮主动脉细胞汇合度诱导的凝血酶生成。将纯化的抗β2糖蛋白 I 抗体孵育在抗磷脂综合征（APS）患者的内皮细胞中。使用血小板无血浆和富含血小板的血浆分别研究内皮细胞中的凝血酶生成和血小板反应性。
内皮细胞汇合度与凝血酶生成呈负相关，而凝血酶生成取决于孵育的 APL 浓度。用 APL 激活内皮细胞可显著增加 PFP 引发的凝血酶生成。PRP 的触发增加了凝血酶谱参数。此外，与血小板孵育的抗β2糖蛋白 I 抗体在 PRP 中放大了凝血酶的形成，并在没有组织因子的情况下诱导血小板激活。
在这项体外研究中，我们证明了在培养的内皮细胞中使用凝血酶生成试验的可行性，并表明需要进行调整以标准化结果。APS 中的促血栓形成状态需要内皮功能障碍和血小板激活。凝血酶形成的定量表明，APL 孵育可诱导培养细胞中的内皮损伤，血小板可放大这种损伤。
