Rosolowsky M, Campbell W B
Department of Pharmacology, University of Texas Southwestern Medical Center, Dallas 75235, USA.
Biochim Biophys Acta. 1996 Jan 19;1299(2):267-77. doi: 10.1016/0005-2760(95)00216-2.
Endothelial cells release several factors which influence vascular tone, leukocyte function and platelet aggregation. Some of these factors are metabolites of arachidonic acid, most notably prostacyclin. However, many of the endothelial metabolites of arachidonic acid have not been positively identified. The purpose of these studies is to identify the arachidonic acid metabolites synthesized by bovine coronary endothelial cells. Cultured bovine coronary artery endothelial cells were incubated with [14C]arachidonic acid. The incubation media was extracted and the radioactive metabolites resolved by a combination of reverse phase- and normal phase-high pressure liquid chromatography (HPLC). The cells synthesized 6-keto prostaglandin (PG)F1 alpha, PGE2, 12-hydroxyheptadecatrienoic acid (HHT), 12-, 15-, and 11-hydroxyeicosatetraenoic acids (HETE), and 14,15-, 11,12-, 8,9-, and 5,6-epoxyeicosatrienoic acids (EET). Several of the HETEs were further analyzed by chiral-phase HPLC. The cells synthesized predominately 12(S)-, 15(S)-, and 11(R)-HETE. The synthesis of the S optical isomers of 12- and 15-HETE suggested that the 12- and 15-lipoxygenases were present in these cells. 11(R)-HETE is probably derived from cyclooxygenase. They also synthesized smaller amounts of 9-, 8- and 5-HETEs. The structures of the HETEs and EETs were confirmed by mass spectrometry. The release of 6-keto PGF1 alpha and 15-HETE was measured by specific radioimmunoassays. Melittin, thrombin, arachidonic acid and A23187 stimulated the release of both eicosanoids in a concentration-related matter. Under all conditions, the release of 6-keto PGF1 alpha exceed the release of 15-HETE. Therefore, cultured bovine coronary artery endothelial cells synthesize cyclooxygenase, lipoxygenase and cytochrome P-450 metabolites of arachidonic acid.
内皮细胞释放多种影响血管张力、白细胞功能和血小板聚集的因子。其中一些因子是花生四烯酸的代谢产物，最显著的是前列环素。然而，许多花生四烯酸的内皮代谢产物尚未得到确切鉴定。这些研究的目的是鉴定牛冠状动脉内皮细胞合成的花生四烯酸代谢产物。将培养的牛冠状动脉内皮细胞与[14C]花生四烯酸一起孵育。提取孵育培养基，并通过反相和正相高压液相色谱（HPLC）相结合的方法分离放射性代谢产物。这些细胞合成了6-酮前列腺素（PG）F1α、PGE2、12-羟基十七碳三烯酸（HHT）、12-、15-和11-羟基二十碳四烯酸（HETE），以及14,15-、11,12-、8,9-和5,6-环氧二十碳三烯酸（EET）。通过手性相HPLC对几种HETE进行了进一步分析。这些细胞主要合成12(S)-、15(S)-和11(R)-HETE。12-和15-HETE的S型光学异构体的合成表明这些细胞中存在12-和15-脂氧合酶。11(R)-HETE可能源自环氧化酶。它们还合成了少量的9-、8-和5-HETE。通过质谱法确认了HETE和EET的结构。通过特异性放射免疫测定法测量6-酮PGF1α和15-HETE的释放。蜂毒肽、凝血酶、花生四烯酸和A23187以浓度相关的方式刺激这两种类花生酸的释放。在所有条件下，6-酮PGF1α的释放量超过15-HETE的释放量。因此，培养的牛冠状动脉内皮细胞合成花生四烯酸的环氧化酶、脂氧合酶和细胞色素P-450代谢产物。
