Ayukawa Osamu, Nakamura Kazuo, Kariyazono Hiroko, Ikeda Ryuji, Arima Junko, Shinkawa Terutoshi, Iwase Harunobu, Sakata Ryuzo, Yamada Katsushi
Department of Clinical Pharmacy and Pharmacology, Graduate School of Medical and Dental Sciences, Kagoshima University, Kagoshima, Japan.
Blood Coagul Fibrinolysis. 2009 Apr;20(3):176-84. doi: 10.1097/MBC.0b013e328322ffd5.
To investigate platelet responsiveness during cold storage of whole blood, we examined platelet aggregation, expression of CD40 ligand (CD40L) on platelets, the plasma levels of soluble form of CD40L (sCD40L) as well as platelet-leukocyte aggregates. Flow cytometry analysis was performed to investigate platelet-leukocyte aggregate formation using antibodies against CD42b and CD45 and platelet activation using antibodies against P-selectin and PAC-1. Blood samples were collected from healthy volunteers, patients with cardiovascular diseases, or both. In the healthy volunteers' blood samples stored at 4 degrees C for 6 h, platelet aggregation in response to 1 micromol/l ADP was enhanced, and released levels of soluble form of P-selectin and thromboxane B2 in response to 1 micromol/l ADP markedly increased. In the samples stored at 4 degrees C for 6 h but not stimulated by any agonists, CD40L expression on the platelets was increased, and plasma levels of sCD40L were also elevated. Under the same condition, the increase in simultaneous expression of CD45 and CD42b was observed. In patients with cardiovascular diseases, the platelet aggregability, coexpression of P-selectin and PAC-1, expression of CD40L on platelets and both CD45-bound and CD42b-bound subsets were all comparable to those of healthy volunteers' samples stored at 4 degrees C for 6 h. Plasma levels of sCD40L in patients were higher than those in healthy volunteers' control. Taken together, storage of whole blood at 4 degrees C for 6 h caused platelet activation comparable to that of patients with cardiovascular diseases, and enhanced platelet activity in such patients may be involved in increased risk for thromboembolic events.
为研究全血冷藏期间血小板的反应性，我们检测了血小板聚集、血小板上CD40配体（CD40L）的表达、可溶性CD40L（sCD40L）的血浆水平以及血小板-白细胞聚集体。采用抗CD42b和CD45抗体通过流式细胞术分析研究血小板-白细胞聚集体的形成，采用抗P-选择素和PAC-1抗体研究血小板活化。血液样本取自健康志愿者、心血管疾病患者或两者。在4℃储存6小时的健康志愿者血液样本中，对1μmol/l ADP的血小板聚集增强，对1μmol/l ADP的可溶性P-选择素和血栓素B2的释放水平显著增加。在4℃储存6小时但未受任何激动剂刺激的样本中，血小板上CD40L的表达增加，sCD40L的血浆水平也升高。在相同条件下，观察到CD45和CD42b同时表达增加。在心血管疾病患者中，血小板聚集性、P-选择素和PAC-1的共表达、血小板上CD40L的表达以及与CD45结合和与CD42b结合的亚群均与在4℃储存6小时的健康志愿者样本相当。患者的sCD40L血浆水平高于健康志愿者对照组。综上所述，全血在4℃储存6小时导致血小板活化与心血管疾病患者相当，此类患者血小板活性增强可能与血栓栓塞事件风险增加有关。
