Eicosanoid-dependent and -independent formation of individual [14C]stearoyl-labelled lysophospholipids in collagen-stimulated human platelets.
Low level collagen activation of platelets is mediated via the release of arachidonic acid (AA) from membrane phospholipids and the formation of thromboxane A2 (TxA2). To assess the specific phospholipids undergoing deacylation via phospholipase A2 thereby providing source(s) of releasable AA, we have measured the individual lysophospholipid formations in platelets prelabelled with [14C]stearic acid and incubated with a low level (2 micrograms/ml) or a high level (10 micrograms/ml) of collagen in the absence or presence of BW755C, a dual inhibitor of cyclooxygenase and lipoxygenase activities. Collagen activation resulted in the generation of [14C]stearoyl-labelled lysophosphatidylinositol (lysoPI), lysophosphatidylcholine (lysoPC), lysophosphatidylethanolamine (lysoPE) and lysophosphatidylserine. BW755C significantly inhibited these collagen-induced changes, suggesting that much of the lysophospholipid, and therefore AA release, was eicosanoid-mediated. At the lower level of collagen, considerable generation of [14C]lysoPE was maintained even in the presence of BW755C, suggesting an eicosanoid-independent degradation of phosphatidyl-ethanolamine. The TxA2-dependent release of AA was also investigated in U-46619-stimulated platelets. This TxA2 mimetic induced considerable formation of the 14C-labelled lysophospholipids, including lysoPI and lysoPC, but not lysoPE. These results suggest that an eicosanoid-independent degradation of phosphatidylethanolamine via phospholipase A2 at lower collagen levels may provide a source of the initial AA for conversion to TxA2 and the subsequent deacylation of phosphatidylinositol, phosphatidylcholine, and also phosphatidylserine.
Thomas
L M
LM
Department of Nutritional Sciences, University of Guelph, Canada.
Holub
B J
BJ
D015772:4,5-Dihydro-1-(3-(trifluoromethyl)phenyl)-1H-pyrazol-3-amine / Q000494:pharmacology; D001792:Blood Platelets / Q000187:drug effects / Q000378:metabolism*; D002250:Carbon Radioisotopes; D003094:Collagen / Q000494:pharmacology*; D006801:Humans; D066298:In Vitro Techniques; D007700:Kinetics; D008246:Lysophospholipids / Q000096:biosynthesis / Q000097:blood* / Q000302:isolation & purification; D011865:Radioisotope Dilution Technique; D013229:Stearic Acids / Q000097:blood*
D002250:Carbon Radioisotopes; D008246:Lysophospholipids; D013229:Stearic Acids; D015772:4,5-Dihydro-1-(3-(trifluoromethyl)phenyl)-1H-pyrazol-3-amine; D003094:Collagen