1,3-Dioctanoylglycerol (1,3-DiC8) is as effective as 1,2-dioctanoylglycerol (1,2-DiC8) in priming phospholipase A2 activation in human platelets and neutrophils.
In the present study, we investigated the effects of different diacylglycerols in comparison with phorbol 12-myristate 13-acetate (PMA) on eicosanoid-independent phospholipase A2 (PLA2) activation in human platelets and neutrophils. Eicosanoid-independent PLA2 activation was measured under conditions where both cyclooxygenase and lipoxygenases were blocked by BW755C. In the presence of PMA (50 nM), the amount of mass arachidonic acid (AA) released represented 400 and 257% of control (without PMA) in A23187-stimulated platelets and neutrophils, respectively, while 1,2-dioctanoylglycerol (1,2-DiC8) and 1-oleoyl-2-acetyl-sn-glycerol (OAG) had increased the eicosanoid-independent AA release by 150 and 117-134% of control, in platelets and neutrophils, respectively. Our results further demonstrate that 1,3-dioctanoylglycerol (1,3-DiC8), a poor activator of protein kinase C (PKC), is nearly as effective as diacylglycerols, such as OAG and 1,2-DiC8 (activators of PKC) in priming PLA2 activation, but is less effective than PMA as a priming agent. However, all three diacylglycerols were less effective than PMA as priming agents. Furthermore, diacylglycerols including 1,3-DiC8 exerted a much greater effect on PLA2 activation in platelets than in neutrophils. Neither 1,3-DiC8 nor 1,2-DiC8 and OAG had any significant priming effect on the accumulation of palmitic and stearic acids, while PMA caused a substantial accumulation of these fatty acids in platelets, but not in neutrophils. We also found that exogenously added OAG underwent significant hydrolysis even in unstimulated platelets, but not in neutrophils, suggesting that exogenously added OAG may be readily accessible for diacylglycerol (DAG) lipase/PLA1 in platelets.(ABSTRACT TRUNCATED AT 250 WORDS)
Murthy
M
M
Department of Pediatrics and Child Health, Manitoba Institute of Cell Biology, University of Manitoba, Winnipeg, Canada.
Rao
G H
GH
Reddy
S
S
D016718:Arachidonic Acid / Q000378:metabolism; D001792:Blood Platelets / Q000201:enzymology*; D000001:Calcimycin / Q000494:pharmacology; D004075:Diglycerides / Q000494:pharmacology*; D004789:Enzyme Activation; D006801:Humans; D009504:Neutrophils / Q000201:enzymology*; D010741:Phospholipases A / Q000378:metabolism*; D054466:Phospholipases A1; D054467:Phospholipases A2; D011493:Protein Kinase C / Q000502:physiology; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
D004075:Diglycerides; C044295:1,2-dioctanoylglycerol; C058807:1,3-dicapryloylglycerol; D016718:Arachidonic Acid; D000001:Calcimycin; C038237:1-oleoyl-2-acetylglycerol; D011493:Protein Kinase C; D010741:Phospholipases A; D054466:Phospholipases A1; D054467:Phospholipases A2; D013755:Tetradecanoylphorbol Acetate