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目的:研究牛心内膜内皮细胞(EEC)腺苷三磷酸双磷酸酶的抗血小板聚集效应。方法:培养牛EEC。以高效液相色谱法测定ADP,用比浊法测血小板聚集。结果:ADP 500μmol·L~(-1)与EEC温孵后,引起ADP浓度进行性地降低。与此相适应的是,未代谢的ADP诱导血小板聚集的能力也降低。在阿司匹林处理过的EEC 1×10~9cells·L~(-1)存在下,凝血酶500U·L~(-1)及PAF 1nmol·L~(-1)诱导的经阿司匹林1mmol·L~(-1)及亚甲基蓝10μmol·L~(-1)处理过的血小板聚集明显受到抑制,且聚集是可逆的;EEC的此种作用类似于腺苷三磷酸双磷酸酶的作用。EEC明显抑制ADP 5μmol·L~(-1)诱导的血小板聚集,但不能抑制ADP-β-S 15μmol·L~(-1)诱导的血小板聚集。结论:腺苷三磷酸双磷酸酶水解ADP是牛EEC重要的抗血栓机制。
Objective: To study the anti-platelet aggregation effect of adenosine triphosphate diphosphatase in bovine endocardial endothelial cells (EEC). Methods: Bovine EECs were cultured. ADP was determined by HPLC and platelet aggregation was measured by turbidimetry. Results: The incubation of ADP with 500 μmol·L -1 with EEC resulted in the progressive decrease of ADP concentration. Consistent with this, the ability of unmetabolized ADP to induce platelet aggregation is also reduced. In the presence of 1 × 10 ~ 9cells · L -1 of EEC treated with aspirin, 500U · L -1 of thrombin and 1nmol·L -1 of PAF induced by aspirin 1mmol·L ~ (-1) -1) and methylene blue (10μmol·L -1) significantly inhibited platelet aggregation and the aggregation was reversible. The effect of EEC was similar to that of adenosine triphosphate phosphatase. EEC significantly inhibited platelet aggregation induced by 5μmol·L -1 ADP but did not inhibit platelet aggregation induced by 15μmol·L -1 ADP-β-S. Conclusion: ADP hydrolysis by ADP is an important antithrombotic mechanism of bovine EEC.