Application of annexin V to measurement of platelete activation

膜联蛋白V在血小板活化测定中的应用

• 批准号: 18591793
• 负责人: SATO Hirokazu
• 金额: $ 2.08万
• 依托单位: Akita University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2006
• 资助国家: 日本
• 起止时间: 2006 至 2007
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18591793/
• 关键词: annexin V; platelete; activation; measurement

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

在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的实验通过肝素采血进行。尽管在实验中用甲醛进行血小板的固定，但发生了甲醛对血小板的活化。然后，认为通过使用甲醛固定来保存血小板是困难的。为此，有必要对妇科恶性肿瘤患者进行血小板不固定化实验，分别采集枸橼酸抗凝血和肝素抗凝血，用Vidas D-dimer 2 '试剂盒测定D-dimer值。肝素采血组D-二聚体值始终高于枸橼酸采血组，且相关系数呈强正相关 ...更多信息 0.998显示。对于妇科恶性肿瘤患者，与健康人相比，血液凝固加速。单用D-二聚体的变化不能反映化疗前后凝血功能的变化。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出现的酸性磷脂的测量程序。在实验中，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。但是，在使用化疗药物的化疗前后，用普通方法测定的血小板聚集能力和用Alexa Fluor 488标记的annexinV检测的酸性磷脂的出现没有变化的趋势。在本研究中，妇科恶性肿瘤患者的凝血也在加速。一般来说，抗血栓药物被认为是血栓形成的原因。然后，在使用抗凝血药物后估计血液凝固的上调。然而，化疗后的血液凝固的明显上调没有检测到这种方法。那么，似乎需要进一步检查。此外，即使使用基于膜联蛋白V的方法，也不能明确血小板聚集能力的变化。虽然在本实验中，血小板上酸性磷脂的出现被用作血小板活化的标志之一，但认为还需要从其他观点出发的方法。少