Pettitt T R, Rowley A F, Barrow S E, Mallet A I, Secombes C J
Biomedical and Physiological Research Group, School of Biological Sciences, University College of Swansea, Singleton Park, United Kingdom.
J Biol Chem. 1991 May 15;266(14):8720-6.
Rainbow trout macrophages maintained in short term culture when incubated with either calcium ionophore, A23187, or opsonized zymosan synthesize a range of lipoxygenase products including lipoxins and leukotrienes. These cells are unusual in that they generate more lipoxin than leukotriene following such challenge. The main lipoxin synthesized was lipoxin (LX) A4. This compound was identified by cochromatography with authentic standard during reversephase high performance liquid chromatography, by ultra violet spectral analysis, radiolabeling following incorporation of [14C]arachidonic acid substrate into macrophage phospholipids, and gas chromatography electron impact mass spectrometry of the methyl ester, trimethylsilyl ether derivative. Other 4-series lipoxins synthesized by trout macrophages were identified as 11-trans-LXA4, 7-cis-11-trans-LXA4, and 6(S)-LXA4. These cells also produced 5-series lipoxins tentatively identified as LXA5, 11-trans-LXA5 and possibly 6(S)-LXA5. No LXB4 or LXB5 was, however, detected. The dynamics of leukotriene and lipoxin release were also determined. Lipoxin generation was slower than leukotriene generation the latter reaching a maximum after 30 min of exposure to ionophore (5 microM, 18 degrees C) compared with 45 min for the former.
在短期培养中,虹鳟巨噬细胞与钙离子载体A23187或调理酵母聚糖一起孵育时,会合成一系列脂氧合酶产物,包括脂oxin和白三烯。这些细胞不同寻常之处在于,在这种刺激后,它们产生的脂oxin比白三烯更多。合成的主要脂oxin是脂oxin(LX)A4。通过反相高效液相色谱法与标准品共色谱分析、紫外光谱分析、将[14C]花生四烯酸底物掺入巨噬细胞磷脂后的放射性标记以及甲酯、三甲基硅醚衍生物的气相色谱电子轰击质谱法,鉴定了该化合物。虹鳟巨噬细胞合成的其他4系列脂oxin被鉴定为11-反式-LXA4、7-顺式-11-反式-LXA4和6(S)-LXA4。这些细胞还产生了5系列脂oxin,初步鉴定为LXA5、11-反式-LXA5,可能还有6(S)-LXA5。然而,未检测到LXB4或LXB5。还测定了白三烯和脂oxin释放的动力学。脂oxin的产生比白三烯慢,后者在暴露于离子载体(5 microM,18摄氏度)30分钟后达到最大值,而前者为45分钟。