Stenson W F, Nickells M W, Atkinson J P
Prostaglandins. 1981 May;21(5):675-89. doi: 10.1016/0090-6980(81)90225-2.
Murine macrophage-like cell lines, J774.2, P388D1, RAW264.7 and PU-5-1R, were incubated with exogenous arachidonic acid (AA). The major metabolites were identified by comigration with known standards in TLC and HPLC and by characteristic behavior following reduction. During a 30 min incubation J774.2 cells metabolized exogenous 14C-AA (10 microM) to PGE2 (14.8%), 12-hydroxy-5,8,10-heptadecatrienoic acid (HHT) (13.0%), thromboxane B2 (TXB2) (7.4%), PGD2 (4.4%) and PGF2 alpha (3.0%). The remainder was incorporated into phospholipids (39.0%), triglycerides (6.1%), and as yet unidentified metabolites (8.2%). No PGF1 alpha was found. Metabolism of exogenous AA was rapid, being less than 90% completed at 3.5 min. Metabolism of exogenous AA is not increased by the simultaneous addition of macrophage stimuli including the cation ionophore A-23187, particulate phagocytic stimuli and endotoxin. The synthesis of cyclooxygenase products was inhibited by low doses of indomethacin (ID50=0.6 microM) while the synthesis of TXB2 and HHT was selectively inhibited by benzylimidazole (ID50=9.5 microM). Identification of a probable lipoxygenase product is being pursued. The synthesis of this product is not inhibited by indomethacin and migrates with an Rf value close to 5,12-diHETE in TLC. P388D1 and RAW264.7 cells metabolize exogenous AA to the same products as J774.2, but in different proportions, while PU-5-1R does not produce cyclooxygenase metabolites to any appreciable extent.
将鼠巨噬细胞样细胞系J774.2、P388D1、RAW264.7和PU - 5 - 1R与外源性花生四烯酸（AA）一起孵育。通过在薄层层析（TLC）和高效液相色谱（HPLC）中与已知标准品共迁移以及还原后的特征行为来鉴定主要代谢产物。在30分钟的孵育过程中，J774.2细胞将外源性14C - AA（10微摩尔）代谢为前列腺素E2（PGE2，14.8%）、12 - 羟基 - 5,8,10 - 十七碳三烯酸（HHT，13.0%）、血栓素B2（TXB2，7.4%）、前列腺素D2（PGD2，4.4%）和前列腺素F2α（PGF2α，3.0%）。其余部分则分别掺入磷脂（39.0%）、甘油三酯（6.1%）以及尚未鉴定的代谢产物（8.2%）中。未发现前列腺素F1α。外源性AA的代谢很快，在3.5分钟时代谢完成不到90%。同时添加包括阳离子离子载体A - 23187、颗粒吞噬刺激物和内毒素在内的巨噬细胞刺激剂，并不会增加外源性AA的代谢。低剂量的吲哚美辛（半数抑制浓度ID50 = 0.6微摩尔）可抑制环氧化酶产物的合成，而苄基咪唑（ID50 = 9.5微摩尔）可选择性抑制TXB2和HHT的合成。正在对一种可能的脂氧合酶产物进行鉴定。该产物的合成不受吲哚美辛抑制，并且在TLC中的比移值（Rf）接近5,12 - 二羟基二十碳四烯酸（diHETE）。P388D1和RAW264.7细胞将外源性AA代谢为与J774.2相同的产物，但比例不同，而PU - 5 - 1R在任何可观程度上都不产生环氧化酶代谢产物。
