Ingerman-Wojenski C, Silver M J, Smith J B, Macarak E
J Clin Invest. 1981 May;67(5):1292-6. doi: 10.1172/jci110157.
Bovine aortic endothelial and smooth muscle cells in culture were incubated with arachidonic acid or prostaglandin H2. The amount of prostacyclin nd thromboxane A2 synthesized ws then determined by specific radioimmunoassay for 6-keto-prostaglandin F1 alpha and thromboxane B2. Although smooth muscle cells produced only 6-keto-prostaglandin F1 alpha and thromboxane B2 in a ratio of 5:1 to 10:1. The same ratio of these metabolites of arachidonic acid ws also found when prostaglandin production from endogenous arachidonic acid was stimulated in endothelial cells by the ionophore A23187. Cyclooxygenase inhibitors inhibited the production of both metabolites equally, whereas thromboxane synthetase inhibitors selectively inhibited the production of thromboxane B2. Cells in culture were also incubated with [1-14C]arachidonic acid and the extracted products were identified by two-dimensional thin-layer chromatography. 6-Keto-prostaglandin F1 alpha was the only metabolite produced by smooth muscle cells, but endothelial cells synthesized 6-keto prostaglandin F1 alpha, thromboxane B2, prostaglandin E2, and prostaglandin F2 alpha.
将培养的牛主动脉内皮细胞和平滑肌细胞与花生四烯酸或前列腺素H2一起孵育。然后通过针对6-酮-前列腺素F1α和血栓素B2的特异性放射免疫测定法来确定合成的前列环素和血栓素A2的量。虽然平滑肌细胞仅产生6-酮-前列腺素F1α和血栓素B2,其比例为5:1至10:1。当用离子载体A23187刺激内皮细胞中内源性花生四烯酸产生前列腺素时,也发现了花生四烯酸这些代谢产物的相同比例。环氧化酶抑制剂同样抑制这两种代谢产物的产生,而血栓素合成酶抑制剂则选择性地抑制血栓素B2的产生。还将培养的细胞与[1-14C]花生四烯酸一起孵育,并通过二维薄层色谱法鉴定提取的产物。6-酮-前列腺素F1α是平滑肌细胞产生的唯一代谢产物,但内皮细胞合成了6-酮-前列腺素F1α、血栓素B2、前列腺素E2和前列腺素F2α。