Laboratory of Molecular Hematology, Department of Medicine, Democritus University of Thrace, 68100 Alexandroupolis, Greece.
Natural Products Synthesis and Bioorganic Chemistry Laboratory, Institute of Nanoscience and Nanotechnology, NCSR "Demokritos", 15310 Athens, Greece.
Int J Mol Sci. 2020 May 21;21(10):3625. doi: 10.3390/ijms21103625.
Neutrophils through the release of neutrophil extracellular traps (NETs) containing active tissue factor (TF) are key components of thrombo-inflammation. Platelets-neutrophils interplay in ST elevation myocardial infarction (STEMI) promotes NET formation via inorganic polyphosphates (polyP) released by thrombin-activated platelets. NETs, however, are also induced by biomaterials in a platelet-independent manner. Considering the possible pleiotropic effects of Ticagrelor beyond platelet inhibition and the clinical need for novel antithrombotic strategies targeting inflammation, we investigated the effects of Ticagrelor on polyP and stent-induced NETs in STEMI. Neutrophils from healthy individuals and patients receiving Ticagrelor were stimulated with polyP or drug-eluting stents (DES) to produce NETs. To induce TF expression, neutrophils were further incubated with plasma obtained from the infarct-related artery (IRA) of STEMI patients. The effects of Ticagrelor on NETs and TF loading were assessed using fluorescence microscopy, flow cytometry, myeloperoxidase(MPO)/DNA complex ELISA, and a Western blot. Ticagrelor interrupts platelet-neutrophil interaction by attenuating NETs induced by polyP. However, Ticagrelor does not affect polyP secretion from thrombin-activated platelets. Similarly, the intracellular production of TF in neutrophils triggered by IRA plasma is not hindered by Ticagrelor. Furthermore, DES induce NETs and synchronous stimulation with IRA plasma leads to the formation of thrombogenic TF-bearing NETs. Ticagrelor inhibits stent-induced NET release. These findings suggest a novel immune-modulatory effect of Ticagrelor when it attenuates the formation of thrombogenic NETs.
中性粒细胞通过释放含有活性组织因子 (TF) 的中性粒细胞细胞外陷阱 (NETs) 是血栓炎症的关键组成部分。血小板-中性粒细胞在 ST 段抬高型心肌梗死 (STEMI) 中的相互作用通过凝血酶激活的血小板释放的无机多聚磷酸盐 (polyP) 促进 NET 形成。然而，NETs 也可以通过血小板独立的方式被生物材料诱导。考虑到替格瑞洛除了抑制血小板以外的可能的多效作用，以及针对炎症的新型抗血栓形成策略的临床需求，我们研究了替格瑞洛对 STEMI 中多聚磷酸盐和支架诱导的 NETs 的影响。用多聚磷酸盐或药物洗脱支架 (DES) 刺激来自健康个体和接受替格瑞洛治疗的患者的中性粒细胞产生 NETs。为了诱导 TF 表达，进一步用 STEMI 患者梗死相关动脉 (IRA) 的血浆孵育中性粒细胞。用荧光显微镜、流式细胞术、髓过氧化物酶 (MPO)/DNA 复合物 ELISA 和 Western blot 评估替格瑞洛对 NETs 和 TF 加载的影响。替格瑞洛通过减弱多聚磷酸盐诱导的 NETs 来中断血小板-中性粒细胞相互作用。然而，替格瑞洛不影响凝血酶激活的血小板分泌多聚磷酸盐。同样，IRA 血浆触发的中性粒细胞内 TF 的产生不受替格瑞洛的阻碍。此外，DES 诱导 NETs，与 IRA 血浆同步刺激会导致形成血栓形成性 TF 携带的 NETs。替格瑞洛抑制支架诱导的 NET 释放。这些发现表明替格瑞洛在减弱血栓形成性 NETs 的形成时具有新的免疫调节作用。
