Kroegel C, Matthys H
Department of Pneumology, University of Freiburg, Germany.
Immunology. 1993 Feb;78(2):279-85.
The spontaneous and stimulated generation of fatty acid cyclo-oxygenase pathway-derived products of arachidonic acid from highly purified (91.6 +/- 1.3%, n = 23) human blood eosinophils obtained from asthmatics were examined using combined gas chromatography/mass spectrometry. Under resting conditions, eosinophils spontaneously generated 0.24 +/- 0.10 pg prostaglandin E2 (PGE2), 0.51 +/- 0.20 prostaglandin D2 (PGD2), 0.35 +/- 0.10 pg prostaglandin F2 alpha (PGF2 alpha) and 8.5 +/- 2.2 pg thromboxane B2 (TXB2), the stable metabolite of TXA2 per 10(6) cells. In contrast, 6-keto-prostaglandin F1 alpha and 9 alpha,11 beta-prostaglandin F2 were not detectable. Stimulation of eosinophils with platelet-activating factor (PAF) for 5 min induced a two- to sixfold increase in the biosynthesis of prostanoids. More than 95% of the generated prostanoids were released into the surrounding medium. The response to PAF was inhibited by the PAF receptor antagonist WEB 2086 (1 microM). The fatty acid cyclo-oxygenase inhibitor, ibuprofen, abolished both the spontaneous and PAF-stimulated generation of prostanoids by eosinophils. LTB4, PMA and calcimycin also produced an increase in prostanoid production, whereas lyso-PAF, the PAF precursor and metabolite, failed to induce prostanoid generation over basal production. In conclusion, the results demonstrate that PAF potently activates human eosinophils to generate and release several fatty acid cyclo-oxygenase metabolites of the arachidonic acid pathway, with TXB2 being the most abundant. These data are in agreement with previous observations suggesting that PAF may be an important stimulus for prostanoid release by the eosinophil in allergic diseases such as asthma.
利用气相色谱/质谱联用技术,检测了从哮喘患者中获取的高度纯化(91.6±1.3%,n = 23)的人血嗜酸性粒细胞中,花生四烯酸的脂肪酸环氧化酶途径衍生产物的自发生成和刺激生成情况。在静息条件下,嗜酸性粒细胞每10⁶个细胞自发生成0.24±0.10 pg前列腺素E2(PGE2)、0.51±0.20 pg前列腺素D2(PGD2)、0.35±0.10 pg前列腺素F2α(PGF2α)和8.5±2.2 pg血栓素B2(TXB2),TXB2是TXA2的稳定代谢产物。相比之下,6-酮-前列腺素F1α和9α,11β-前列腺素F2无法检测到。用血小板活化因子(PAF)刺激嗜酸性粒细胞5分钟,可使前列腺素生物合成增加2至6倍。超过95%生成的前列腺素释放到周围介质中。PAF受体拮抗剂WEB 2086(1 microM)可抑制对PAF的反应。脂肪酸环氧化酶抑制剂布洛芬可消除嗜酸性粒细胞自发和PAF刺激的前列腺素生成。白三烯B4(LTB4)、佛波酯(PMA)和离子霉素也会使前列腺素生成增加,而PAF前体和代谢产物溶血PAF未能诱导前列腺素生成超过基础生成量。总之,结果表明PAF能有效激活人嗜酸性粒细胞,生成并释放花生四烯酸途径的几种脂肪酸环氧化酶代谢产物,其中TXB2最为丰富。这些数据与先前的观察结果一致,表明PAF可能是哮喘等过敏性疾病中嗜酸性粒细胞释放前列腺素的重要刺激因素。