Defontis Myriam, Côté Serge, Stirn Martina, Ledieu David
Department of Veterinary Clinical Sciences, Clinical Pathology and Clinical Pathophysiology, Justus-Liebig University Giessen, Frankfurterstrasse 126, 35392 Giessen, Germany.
Exp Toxicol Pathol. 2013 Jul;65(5):637-44. doi: 10.1016/j.etp.2012.07.003. Epub 2012 Aug 10.
This study was performed to optimize and standardize the use of the Multiplate(®) whole blood impedance aggregometer in the Beagle dog and Wistar rat for use in a research laboratory environment. The anticoagulants citrate, heparin and hirudin were compared and platelet aggregation responses to ADP, collagen, arachidonic acid and Par-4 agonist were evaluated to determine their half maximal effective concentrations (EC(50)) in blood containing low concentrations of a drug solvent (0.1% DMSO). The results indicate that citrate anticoagulation is not suitable for Multiplate(®) whole blood aggregometry because of the presence of spontaneous aggregation. ADP and collagen were found to be appropriate agonists for both species, whereas in the Beagle dog Par-4 agonist failed to induce aggregation and arachidonic acid induced platelet aggregation showed a high interindividual variability. The agonists EC(50) calculated in hirudin blood were 2.70 μM ADP, 0.85 μg/ml collagen, 0.03 mM arachidonic acid and 165.7 μM Par-4 agonist in the Wistar rat, and 0.95 μM ADP and 0.23 μg/ml collagen in the Beagle dog.
本研究旨在优化并规范在比格犬和Wistar大鼠中使用Multiplate®全血阻抗聚集仪，以便在研究实验室环境中应用。比较了抗凝剂柠檬酸盐、肝素和水蛭素，并评估了血小板对ADP、胶原蛋白、花生四烯酸和Par-4激动剂的聚集反应，以确定它们在含有低浓度药物溶剂（0.1% DMSO）的血液中的半数最大效应浓度（EC50）。结果表明，由于存在自发聚集，柠檬酸盐抗凝不适用于Multiplate®全血聚集测定。发现ADP和胶原蛋白是两种动物合适的激动剂，而在比格犬中，Par-4激动剂未能诱导聚集，花生四烯酸诱导的血小板聚集表现出较高的个体间变异性。在Wistar大鼠中，水蛭素抗凝血中计算出的激动剂EC50为2.70 μM ADP、0.85 μg/ml胶原蛋白、0.03 mM花生四烯酸和165.7 μM Par-4激动剂，在比格犬中为0.95 μM ADP和0.23 μg/ml胶原蛋白。
