Mass determination of receptor-mediated accumulation of phosphatidate and diglycerides in human neutrophils measured by Coomassie blue staining and densitometry.
Mass determination of receptor-mediated accumulation of phosphatidate and diglycerides in human neutrophils measured by Coomassie blue staining and densitometry.
复制标题
通过考马斯蓝染色和光密度测定法测定人中性粒细胞中受体介导的磷脂酸和甘油二酯的积累。
DOI：
10.1016/0006-291x(89)92407-8
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发表时间：
1989
影响因子：
3.1
通讯作者：
McCall,CE
中科院分区：
文献类型：
--
作者：
Agwu,DE;McPhail,LC;Wykle,RL;McCall,CE
摘要用考马斯蓝染色和密度计量法测定了1-O-[~H]烷基-2-酰基-GPC标记的中性粒细胞在1um fMLP刺激下，1，2-二酰基甘油(AAG)、1-O-烷基-2-酰甘油(EAG)和磷脂酸(PA)的含量。在5s时，AAG和PA分别比对照组增加80%和107%。PA的积累在30s时达到最大值，5s时比AAG高302%，30s时高550%。刺激中性粒细胞后，EAG蓄积延迟15s。这些结果表明，AAG在EAG之前积累，并支持AAG在细胞激活中的作用，可能是通过刺激蛋白激酶C(PKC)。EAG可能会用来对抗AAG的影响，也可能会引起反应。早期积累的高浓度PA提示，PA可能是通过激活fMLP刺激的PMN中的磷脂酶D而产生的。
Abstract Quantitation of 1, 2-diacylglycerol (AAG), 1-O-alkyl-2-acylglycerol (EAG) and phosphatidic acid (PA) was conducted in polymorphonuclear leukocytes (PMN) labeled with 1-O-[3 H] alkyl-2-acyl-GPC following stimulation with 1 uM fMLP using Coomassie blue staining and densitometry. At 5s AAG and PA increased by 80% and 107%, respectively, over controls. The accumulation of PA, which reached a maximum by 30s, was higher than AAG by 302% at 5s, and 550% at 30s. EAG accumulation was delayed by 15s following stimulation of PMN. These results show that AAG accumulates before EAG and support the role of AAG in cellular activation, perhaps, via the stimulation of protein kinase C (PKC). EAG may serve to counter the effects of AAG or may itself elicit responses. The high concentrations of PA which accumulate early suggest that PA may be generated by the activation of phospholipase D in PMN stimulated with fMLP.