Center for Molecular and Vascular Biology, University Hospital Leuven, KULeuven, Leuven, Belgium.
J Proteomics. 2011 May 16;74(6):902-13. doi: 10.1016/j.jprot.2011.03.007. Epub 2011 Mar 23.
Eight patients with clinical bleeding problems have evidence for platelet storage pool disease as they present with impaired platelet aggregation and secretion with low concentrations of ADP and collagen and an absence of second phase aggregation with epinephrine. Electron microscopy analysis further showed a reduced but not absent amount of platelet dense granules, and CD63 staining was decreased compared to healthy controls. The presence of alpha granules and CD62P expression after platelet activation was normal. This work aimed at identifying differentially expressed proteins in the platelet releasate and its remaining pellet after activation with A23187 and TRAP in patients and controls using DIGE-based proteomic technology. We identified 44 differentially expressed proteins in patients and the altered expression for some of them was confirmed by immunoblot analysis. Most of these proteins belong to the class of cytoskeleton-related proteins. In addition, 29 cytoskeleton-related genes showed an altered expression in platelet mRNA from patients using a real-time PCR array. In conclusion, our study shows that the dense granule secretion defect in patients with platelet storage pool disease is highly heterogeneous with evidence of an underlying cytoskeleton defect.
8 例有临床出血问题的患者存在血小板贮存池疾病的证据，表现为血小板聚集和分泌功能受损，ADP 和胶原浓度降低，肾上腺素诱导的二期聚集缺失。电子显微镜分析进一步显示血小板致密颗粒数量减少，但并未完全缺失，与健康对照组相比 CD63 染色减少。血小板活化后α颗粒和 CD62P 的表达正常。本研究旨在采用 DIGE 蛋白质组学技术，在患者和对照者的血小板释放物及其经 A23187 和 TRAP 激活后的剩余颗粒中，鉴定出有差异表达的蛋白质。我们在患者中鉴定出 44 种有差异表达的蛋白质，其中一些蛋白质的改变表达通过免疫印迹分析得到了证实。这些蛋白质大多属于细胞骨架相关蛋白。此外，通过实时 PCR 阵列，我们在患者的血小板 mRNA 中发现 29 种细胞骨架相关基因的表达发生改变。总之，我们的研究表明，血小板贮存池疾病患者的致密颗粒分泌缺陷具有高度异质性，存在潜在的细胞骨架缺陷。
