Walsh C E, Dechatelet L R, Chilton F H, Wykle R L, Waite M
Biochim Biophys Acta. 1983 Jan 7;750(1):32-40. doi: 10.1016/0005-2760(83)90201-1.
Previous studies have demonstrated that [3H]arachidonic acid is released from prelabeled human neutrophil phospholipids when the cells are stimulated by calcium ionophore A23187 or by opsonized zymosan. Neither lysophospholipid generated by phospholipase A2 activity, diacylglycerol nor monoacylglycerol produced via phospholipase C/diacylglycerol lipase action have been identified following neutrophil challenge. The inability to detect any intermediates during the release of arachidonate is due to either rapid reacylation of lysophospholipid or conversion of diacylglycerol (monoacylglycerol) to cellular acylglycerols. The addition of exogenous [14C]fatty acid at the time of challenge was employed to determine the involvement of either phospholipase A2 or phospholipase C activities. Neutrophil stimulation with calcium ionophore A23187 resulted in an incorporation of exogenous [14C]arachidonate into phosphatidylinositol and phosphatidylcholine, those phospholipids which specifically release arachidonate. When the saturated fatty acid, [14C]stearate, replaced [14C]arachidonate, very little [14C]fatty acid was incorporated into any of the phospholipid species. Lipid phosphorus measurements revealed no significant mass change in any phospholipid class following ionophore challenge. Production of [14C]phosphatidic acid was not detected, as would be expected if diacylglycerol kinase and de novo phospholipid metabolism were significantly involved.
先前的研究表明，当细胞受到钙离子载体A23187或调理酵母聚糖刺激时，[3H]花生四烯酸会从预先标记的人中性粒细胞磷脂中释放出来。在中性粒细胞受到刺激后，未发现磷脂酶A2活性产生的溶血磷脂、磷脂酶C/二酰基甘油脂肪酶作用产生的二酰基甘油或单酰基甘油。在花生四烯酸释放过程中无法检测到任何中间体，这是由于溶血磷脂的快速再酰化或二酰基甘油（单酰基甘油）向细胞酰基甘油的转化。在刺激时添加外源性[14C]脂肪酸用于确定磷脂酶A2或磷脂酶C活性的参与情况。用钙离子载体A23187刺激中性粒细胞会导致外源性[14C]花生四烯酸掺入磷脂酰肌醇和磷脂酰胆碱，这些磷脂会特异性释放花生四烯酸。当饱和脂肪酸[14C]硬脂酸取代[14C]花生四烯酸时，很少有[14C]脂肪酸掺入任何磷脂种类中。脂质磷测量显示，在离子载体刺激后，任何磷脂类别的质量均无显著变化。未检测到[14C]磷脂酸的产生，而如果二酰基甘油激酶和从头磷脂代谢显著参与，则预期会检测到。
