Mike I, Ichikawa Y, Shimizu T, Honda Z I, Seyama Y, Ishii A
Pharmaceutical Research Laboratories, Kyowa Hakko Kogyo Co., Ltd., Shizuoka, Japan.
Prostaglandins. 1991 Nov;42(5):431-40. doi: 10.1016/0090-6980(91)90034-d.
Anti-12(S)-hydroxyeicosatetraenoic acid (12-HETE)-antibody and anti-thromboxane B2 (TXB2)-antibody were generated and applied to the radioimmunoassay. The detection limit for 12-HETE was 16 pg. The cross-reactivities of anti-12-HETE-antibody were 4.6% for 15-HETE, 0.18% for 5-HETE and below 0.15% for leukotrienes and prostaglandins (PGs). 12-HETE and TXB2 released from guinea pig platelets were measured by radioimmunoassay. Platelet activating factor (PAF) at 10(-9) M induced the aggregation of platelets, the releases of immunoreactive-12-HETE (1.8 +/- 1.2 ng/10(8) platelets, mean +/- S.D.) and immunoreactive-TXB2 (18.5 +/- 17.3 ng/10(8) platelets). Collagen at 1 microgram/ml also evoked platelet aggregation, the releases of immunoreactive-12-HETE (2.7 +/- 1.1 ng/10(8) platelets) and immunoreactive-TXB2 (11.8 +/- 4.6 ng/10(8) platelets). By the stimulation with these compounds, TXB2 was produced in a greater amount than 12-HETE from guinea pig platelets. Although 10(-7) M and 10(-6) M U46619, a TXA2 mimetic, caused platelet aggregation, arachidonic acid metabolites were not released. These data suggest the presence of different mechanisms of platelet activation depending on each stimulus.
制备了抗12（S）-羟基二十碳四烯酸（12-HETE）抗体和抗血栓素B2（TXB2）抗体，并将其应用于放射免疫分析。12-HETE的检测限为16皮克。抗12-HETE抗体对15-HETE的交叉反应性为4.6%，对5-HETE为0.18%，对白三烯和前列腺素（PGs）低于0.15%。通过放射免疫分析测定豚鼠血小板释放的12-HETE和TXB2。10^(-9) M的血小板活化因子（PAF）诱导血小板聚集、免疫反应性12-HETE（1.8±1.2纳克/10^8个血小板，平均值±标准差）和免疫反应性TXB2（18.5±17.3纳克/10^8个血小板）的释放。1微克/毫升的胶原蛋白也引起血小板聚集、免疫反应性12-HETE（2.7±1.1纳克/10^8个血小板）和免疫反应性TXB2（11.8±4.6纳克/10^8个血小板）的释放。通过这些化合物刺激，豚鼠血小板产生的TXB2量比12-HETE多。尽管10^(-7) M和10^(-6) M的TXA2模拟物U46619引起血小板聚集，但花生四烯酸代谢产物未释放。这些数据表明，根据每种刺激，血小板活化存在不同机制。
