Kirchmaier C M, Altorjay I, Bellinger O, Breddin H K
Abteilung für Angiologie, Zentrum der Inneren Medizin, J. W. Goethe-Universität, Frankfurt/Main.
Z Kardiol. 1991;80 Suppl 5:13-6.
A new method has been developed to investigate the direct effect of endothelial cells on platelet aggregation in the presence of cultured confluent human EC monolayers. The inside of the disk shaped cuvettes are covered by human umbilical vein ECs. The cuvettes rotate in the light beam of a photometer. In these cuvettes the effect of different aggregation inducers was inhibited in a concentration-dependent manner, e. g. for collagen at 0.5 microgram/ml, ADP at 5 x 10(-7) M, epinephrine at 5 x 10(-7) M and thrombin at 0.05 U/ml final concentration. (Platelet count 5 x 10(5)/microliters). Higher concentrations of the inducers led to irreversible platelet aggregation and were used for testing of antiplatelet drugs. In this system we detected a synergism between the antiaggregatory effect of the EC monolayer and that of SIN 1 (Cassella, Frankfurt/Main) the main metabolite of Molsidomine. 10 micrograms/ml PRP of SIN 1 alone partially inhibited platelet aggregation induced by 1 microgram/ml collagen and 10(-6) M ADP respectively in uncovered aggregation cuvettes. In the presence of an EC monolayer complete inhibition of platelet aggregation was obtained at a 5-fold final concentration of both inducers. After pretreatment of ECs with 1 mmol ASA over 30 min. the antiaggregatory effect of SIN 1 decreased, but was more pronounced (50%) than in uncovered cuvettes (25%).
已开发出一种新方法，用于研究在培养的汇合人内皮细胞单层存在的情况下内皮细胞对血小板聚集的直接影响。圆盘形比色皿内部覆盖有人脐静脉内皮细胞。比色皿在光度计的光束中旋转。在这些比色皿中，不同聚集诱导剂的作用以浓度依赖的方式受到抑制，例如，对于胶原蛋白，终浓度为0.5微克/毫升；对于ADP，为5×10⁻⁷M；对于肾上腺素，为5×10⁻⁷M；对于凝血酶，为0.05U/毫升。（血小板计数为5×10⁵/微升）。更高浓度的诱导剂会导致不可逆的血小板聚集，并用于抗血小板药物的测试。在该系统中，我们检测到内皮细胞单层的抗聚集作用与莫索尼定的主要代谢产物SIN 1（卡塞拉公司，美因河畔法兰克福）的抗聚集作用之间存在协同作用。单独使用10微克/毫升的SIN 1富血小板血浆分别在未覆盖的聚集比色皿中部分抑制了由1微克/毫升胶原蛋白和10⁻⁶M ADP诱导的血小板聚集。在内皮细胞单层存在的情况下，两种诱导剂的终浓度为5倍时可完全抑制血小板聚集。用1毫摩尔阿司匹林预处理内皮细胞30分钟后，SIN 1的抗聚集作用降低，但比未覆盖的比色皿中更明显（50%对25%）。
