UBFC, University of Burgundy Franche-Comte, 25000, Besancon, France; EA 3920, University of Franche-Comté, 25000, Besancon, France.
UBFC, University of Burgundy Franche-Comte, 25000, Besancon, France; Plateforme de Lipidomique, INSERM ULR 1231, University of Burgundy Franche-Comte, 21000 Dijon, France.
Chem Phys Lipids. 2018 Nov;216:25-29. doi: 10.1016/j.chemphyslip.2018.09.008. Epub 2018 Sep 14.
Platelets are major targets for the treatment of thrombo-embolic disorders. Their plasma membrane contains specialized microdomains enriched in sphingomyelins and free cholesterol including membrane receptors. P2Y12 receptors need to be situated in these domains to be able to conduct activation signaling by adenosine diphosphate (ADP). We studied the impact of ticagrelor, a P2Y12 antagonist, and ADP on the composition and distribution of sphingomyelins in detergent-resistant membrane (DRM) of platelet membranes. Platelets were obtained from healthy donors. DRMs of platelet membranes were isolated in 4 experimental groups: control; ADP, with platelets stimulated by 20 μM ADP and 5 mM CaCl2; ticagrelor, with platelets incubated by ticagrelor 4 μM methanol dissolved; and ticagrelor + ADP, with incubation by ticagrelor followed by stimulation by ADP as above. After mass spectrometry analysis, we found 16 species of sphingomyelins in platelet membrane DRMs. We also found that treatment with ticagrelor and stimulation by ADP could induce changes in the composition, distribution and concentration of sphingomyelins in membranes of platelets. In all groups, the predominant species of sphingomyelins in platelet membrane was d18:1/16:0. Taken together, our results show that stimulation by ADP or inhibition by ticagrelor changed the level and composition of sphingomyelins in platelet membranes. These changes might be considered as reorganization or new recruitment of certain types of sphingomyelins through the membrane.
血小板是治疗血栓栓塞性疾病的主要靶点。它们的质膜含有富含鞘磷脂和游离胆固醇的特殊微区，包括膜受体。P2Y12 受体需要位于这些区域，才能通过二磷酸腺苷 (ADP) 进行激活信号传导。我们研究了 P2Y12 拮抗剂替格瑞洛和 ADP 对血小板质膜去污剂抗性膜 (DRM) 中鞘磷脂组成和分布的影响。血小板从健康供体中获得。在 4 个实验组中分离血小板质膜的 DRM：对照组；ADP，用 20μM ADP 和 5mM CaCl2 刺激的血小板；替格瑞洛，用 4μM 甲醇溶解的替格瑞洛孵育的血小板；和替格瑞洛+ADP，先用替格瑞洛孵育，然后如上所述用 ADP 刺激。进行质谱分析后，我们在血小板质膜 DRM 中发现了 16 种鞘磷脂。我们还发现，替格瑞洛治疗和 ADP 刺激可诱导血小板质膜中鞘磷脂的组成、分布和浓度发生变化。在所有组中，血小板质膜中鞘磷脂的主要种类为 d18:1/16:0。总之，我们的结果表明，ADP 刺激或替格瑞洛抑制改变了血小板质膜中鞘磷脂的水平和组成。这些变化可能被认为是通过质膜对某些类型的鞘磷脂进行重新排列或新的募集。
