Dobson P, Aharony D, Krell R D
Res Commun Chem Pathol Pharmacol. 1983 Oct;42(1):3-23.
Washed rat peritoneal cells (RPC) rapidly and efficiently incorporated exogenous [3H]-arachidonic acid (AA). Exposure of labeled RPC to the calcium ionophore A23187 induced production of [3H]-leukotriene C4, D4 and E4 (LTC4, LTD4 and LTE4) and [3H]-prostaglandins (PGs). The radiolabeled lipoxygenase metabolites were isolated by a combination of organic extraction, silicic acid chromatography and reverse-phase high pressure liquid chromatography (RP-HPLC). Authentic leukotrienes (LT) were used to identify the biologically-synthesized products on RP-HPLC and to calculate recovery. The endogenously generated LT were also characterized by their ability to contract guinea-pig ileum and their susceptibility to soybean lipoxygenase. A23187 induced a bell-shaped concentration-dependent release of [3H]-LT which peaked at 1 X 10(-6)M ionophore. Kinetic studies revealed that ionophore stimulated the formation of mainly [3H]-LTE4 with only transient accumulation of [3H]-LTC4 and D4. In the presence of cysteine, however, production of [3H]-LTE4 was abolished with a subsequent accumulation of [3H]-LTD4. Non-steroidal antiinflammatory drugs (NSAID) moderately enhanced A23187-induced [3H]-LT production, whereas phospholipase and lipoxygenase inhibitors inhibited. The method is sensitive, reliable, and efficient for measurement of small quantities of LT produced from isolated cells.
洗涤过的大鼠腹膜细胞（RPC）能快速且高效地摄取外源性[3H] - 花生四烯酸（AA）。将标记的RPC暴露于钙离子载体A23187会诱导产生[3H] - 白三烯C4、D4和E4（LTC4、LTD4和LTE4）以及[3H] - 前列腺素（PGs）。通过有机萃取、硅酸色谱和反相高压液相色谱（RP - HPLC）相结合的方法分离放射性标记的脂氧合酶代谢产物。使用 authentic 白三烯（LT）在RP - HPLC上鉴定生物合成产物并计算回收率。内源性产生的LT还通过其使豚鼠回肠收缩的能力及其对大豆脂氧合酶的敏感性来表征。A23187诱导了[3H] - LT呈钟形的浓度依赖性释放，在1×10（-6）M离子载体时达到峰值。动力学研究表明，离子载体主要刺激[3H] - LTE4的形成，而[3H] - LTC4和D4只是短暂积累。然而，在半胱氨酸存在的情况下，[3H] - LTE4的产生被消除，随后[3H] - LTD4积累。非甾体抗炎药（NSAID）适度增强A23187诱导的[3H] - LT产生，而磷脂酶和脂氧合酶抑制剂则起抑制作用。该方法对于测量从分离细胞中产生的少量LT敏感、可靠且高效。 （注：“authentic”这里可能是想说“标准的、正品的”等意思，但在文本中表述不太清晰准确，可能影响对整体内容的理解。）
