Application of annexin V to measurement of platelete activation

EDTA或柠檬酸钠采血时，钙离子发生螯合作用，干扰了膜联蛋白V与血小板表面酸性磷脂的结合。因此，使用膜联蛋白V的实验是通过肝素采血进行的。虽然在实验中用甲醛固定了血小板，但却发生了血小板被甲醛激活的情况。当时，人们认为用甲醛固定保存血小板是困难的。因此，有必要提出一个不固定血小板的实验。对妇科恶性肿瘤患者进行柠檬酸盐采血和肝素采血，采用Vidas检测试剂盒d -二聚体2′测定d -二聚体值。在肝素采血中，d -二聚体的值始终高于柠檬酸盐采血，且相关系数为0.998以上，呈强正相关。妇科恶性肿瘤患者与健康人相比，凝血速度加快。仅通过d -二聚体的变化无法检测抗肿瘤药物化疗前后的凝血变化。价值。采用Alexa Fluor 488标记的酸性磷脂结合蛋白膜联蛋白V建立了活化血小板表面酸性磷脂的测定方法。实验中，Alexa Fluor 488标记的膜联蛋白V比FTTC标记的膜联蛋白V具有更高的重现性。妇科恶性肿瘤患者血小板聚集能力低于健康人。然而，在使用抗肿瘤药物化疗前后，常规方法测量的血小板聚集能力和Alexa Fluor 488标记的annexinV检测的酸性磷脂外观均未观察到。在本研究中，我们发现妇科恶性肿瘤患者的凝血也在加速。一般认为抗肿瘤药物是引起血栓形成的原因。然后，估计使用抗肿瘤药物后凝血功能的上调。然而，该方法未发现化疗后凝血功能明显上调。那么，似乎需要进一步的检查。此外，即使采用基于膜联蛋白v的方法，也无法明确血小板聚集能力的变化。本实验虽然将血小板上酸性磷脂的出现作为血小板活化的标志之一，但认为还需要从其他角度入手。少

In blood collecting using EDTA or sodium citrate, the chelate of the calcium ion occured so that the binding of annexin V to acidic phospholipid on the surface of blood platelete was interfared. For this reason, the experiments using annexin V were conducted by heparin blood collecting. Although fixation of blood platelets was carried out with formaldehyde for the experiment, activation of the blood platelets by formaldehyde took place. Then, it was thought that preservation of the blood platelets by the fixation using formaldehyde was difficult. Therefoe it was necessary to present an experiment with blood platelets without fixation.Citrate blood collecting and heparin blood collecting were performed from the gynecology malignant tumor patients, and the D-dimer value was measured using the Vidas assay kit D-dimer 2'. In heparin blood collecting, the value of D-dimer was always higher than that in citrate blood collection, and strong positive correlation of the correlation coefficient … More 0.998 was shown. As for the gynecology malignant tumor patient, blood coagulation was accelerated as compared with the healthy person. The change of blood coagulation before and after the chemotherapy by an antineoplastic drug was not able to be detected only by change of D-dimer. Value.The measurement procedure of acidic phospholipid appearing on the activated blood platelet surface was established using Alexa Fluor 488 labeled acidic phospholipid binding protein annexin V. In the experiment, the Alexa Fluor 488 labeled annexin V had higher reproducibility than the FTTC labeled one. Platelet aggregation ability was lower in gynecology malignant tumor patients than in healthy persons. However, no tendancy of platelet aggregation ability measured by common procedures and acidic phosphatide appearance detected by Alexa Fluor 488 labeled annexinV was observed before and after the chemotherapy by an antineoplastic drug at this time.In this research, it became clear that blood coagulation was accelerating also in gynecology malignant tumor patients. Generally an antineoplastic drug is considered to be the cause of thrombosis indecements. Then, upregulation of the blood coagulation is estimated after the use of an antineoplastic drug. However, apparent upregulation of blood coagulation after chemotherapy was not detected by this method. Then, it seems that the further examination is required. Moreover, even if the annexin V-based method was used, change of platelet aggregation ability was not able to be clarified. Although the appearance of acidic phospholipid on blood platelete was used as one of the marker of platelete activation in this experiment, it is thought that the approach from other viewpoints was also required. Less