Application of annexin V to measurement of platelete activation

在用EDTA或柠檬酸钠采血时，会发生钙离子的络合作用，从而干扰膜联蛋白V与血小板表面酸性磷脂的结合。为此，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，但也发生了甲醛对血小板的激活。当时，人们认为用甲醛固定保存血小板是困难的。采集妇科恶性肿瘤患者的血样，用Vidas试剂盒D-二聚体2‘测定D-二聚体的值。肝素采血时D-二聚体始终高于柠檬酸盐血采血值，且相关系数…呈显著正相关超过0.998的影片上映了。妇科恶性肿瘤患者与健康人比较，凝血功能加快。单凭D-二聚体的变化不能反映化疗前后凝血功能的变化。建立了用Alexa Fluor 488标记的酸性磷脂结合蛋白Annexin V测定活化的血小板表面酸性磷脂的方法。在实验中，Alexa Fluor 488标记的Annexin V的重复性比FTTC标记的高。妇科恶性肿瘤患者的血小板聚集率低于健康人。然而，用常规方法检测的血小板聚集能力和Alexa Fluor 488标记的膜联蛋白V检测到的酸性磷脂形态在此时抗肿瘤药物化疗前后没有变化。一般来说，抗肿瘤药物被认为是血栓形成的原因。然后，在使用抗肿瘤药物后，评估凝血功能的上调。然而，该方法未检测到化疗后明显的凝血功能上调。那么，似乎还需要进一步的检查。而且，即使采用以膜联蛋白V为基础的方法，也不能阐明血小板聚集能力的变化。本实验虽然将血小板表面酸性磷脂的出现作为血小板活化的标志之一，但也认为还需要从其他角度进行探讨。较少

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