差示扫描量热法测定血小板聚集的热分析。

José F Rivas-Vilchis, Fernando Hernández Sánchez, Rodolfo Velasco Lezama
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摘要

量热法是一种测量热源和样品之间热流的分析方法。样品根据其物理或化学组成获得或损失热量。差示扫描量热法(DSC)将加热样品的结果与加热参考物质的结果进行比较。DSC然后测量内能或样品的热容量。本研究的目的是研究血小板活化的热特性。通过静脉穿刺从人类志愿者身上获得血液,并在5ml硅化和柠檬酸化真空管中收集。血小板计数用血细胞计测定。离心获得富血小板血浆(PRP)或贫血小板血浆(PPP)。将10微升的PRP或PPP放入铝锅中,用肾上腺素(5.0微米)或CaCl2(50微米)激活或不激活进行DSC。为了避免血小板自发活化,将样品冷冻保存,稳定5分钟后,加入5微升凝集诱导剂。稳定后在-12℃下开始扫描,增加5℃/min至最高60℃。实验在TA差示扫描量热计(New Castle, DE, USA)上进行。在血小板活化过程中,PRP和PPP的热演化差异为253 J/g。PRP与PPP激活的热流差异可能对应于血小板聚集的放热过程。
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Thermal analysis of platelet aggregation assessed by differential scanning calorimetry.

Calorimetry is an analytical method that measures heat flow between a heat source and sample. The sample gains or losses heat based on physical or chemical composition. Differential scanning calorimetry (DSC) compares the results of heating a sample to those for heating a reference material. DSC then measures internal energy or a sample's calorific capacity. The aim of this study was to examine the thermal characteristics of platelet activation. Blood was obtained from human volunteers by venipuncture and collected in 5 ml siliconised and citronated vacutainer tubes. Platelet counts were measured using a hemocytometer. Platelet-rich (PRP) or platelet-poor plasma (PPP) was obtained by centrifugation. Ten microliters of PRP or PPP were placed into aluminum pans for DSC with or without activation by epinephrine (5.0 microM) or CaCl2 (50 microM). To avoid a spontaneous activation of platelets samples were kept frozen, after a 5 min period of stabilization, 5 microl of aggregation-inducing agent was added. Scans were initiated at a -12 degrees C after stabilization, with an increase of a 5 degrees C/min to a maximum of 60 degrees C. The experiments were performed on a TA Differential Scanning Calorimeter (New Castle, DE, USA). The difference in heat evolved between the PRP and PPP during the process of platelet activation was 253 J/g. The difference of heat flow in the activation of PRP versus PPP may correspond to an exothermic process involved in platelet aggregation.

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