Nijssen J G, Oosting R S, Nÿkamp F P, van den Bosch H
Lipids. 1986 Oct;21(10):623-8. doi: 10.1007/BF02537210.
Guinea pig alveolar macrophages were labeled by incubation with either arachidonate or linoleate. Arachidonate labeled phosphatidylcholine (PC), phosphatidylethanolamine (PE) and triglycerides (TG) equally well, with each lipid containing about 30% of total cellular radioactivity. In comparison to arachidonate, linoleate was recovered significantly less in PE (7%) and more in TG (47%). To investigate whether redistributions of acyl chains among lipid classes took place, the macrophages were incubated with 1-acyl-2-[1-14C]arachidonoyl PC or 1-acyl-2-[1-14C]linoleoyl PC. After harvesting, the cells incubated with 1-acyl-2-[1-14C]linoleoyl PC contained 86% of the recovered cellular radioactivity in PC, with only small amounts of label being transferred to PE and TG (3 and 6%, respectively). More extensive redistributions were observed with arachidonate-labeled PC. In this case, only 60% of cellular radioactivity was still associated with PC, while 22 and 12%, respectively, had been transferred to PE and TG. Arachidonate transfer from PC to PE was unaffected by an excess of free arachidonate which inhibited this transfer to TG for over 90%, indicating that different mechanisms or arachidonoyl CoA pools were involved in the transfer of arachidonate from PC to PE and TG. Cells prelabeled with 1-acyl-2-[1-14C]arachidonoyl PC released 14C-label into the medium upon further incubation. This release was slightly stimulated by zymosan and threefold higher in the presence of the Ca2+-ionophore A23187. Labeling of macrophages with intact phospholipid molecules appears to be a suitable method for studying acyl chain redistribution and release reactions.
豚鼠肺泡巨噬细胞通过与花生四烯酸盐或亚油酸盐孵育进行标记。花生四烯酸盐能同样良好地标记磷脂酰胆碱(PC)、磷脂酰乙醇胺(PE)和甘油三酯(TG),每种脂质约占细胞总放射性的30%。与花生四烯酸盐相比,亚油酸盐在PE中的回收率显著降低(7%),而在TG中的回收率更高(47%)。为了研究脂质类别之间酰基链的重新分布是否发生,巨噬细胞与1-酰基-2-[1-14C]花生四烯酰基PC或1-酰基-2-[1-14C]亚油酰基PC一起孵育。收获后,用1-酰基-2-[1-14C]亚油酰基PC孵育的细胞中,86%的回收细胞放射性存在于PC中,只有少量标记转移到PE和TG中(分别为3%和6%)。用花生四烯酸盐标记的PC观察到更广泛的重新分布。在这种情况下,只有60%的细胞放射性仍与PC相关,而分别有22%和12%转移到了PE和TG中。花生四烯酸盐从PC转移到PE不受过量游离花生四烯酸盐的影响,而过量游离花生四烯酸盐对其转移到TG的抑制率超过90%,这表明花生四烯酸盐从PC转移到PE和TG涉及不同的机制或花生四烯酰辅酶A池。预先用1-酰基-2-[1-14C]花生四烯酰基PC标记的细胞在进一步孵育时会将14C标记释放到培养基中。这种释放受到酵母聚糖的轻微刺激,在存在Ca2+离子载体A23187的情况下高出三倍。用完整磷脂分子标记巨噬细胞似乎是研究酰基链重新分布和释放反应的合适方法。