Platelet-activating factor preferentially stimulates the phospholipase A2/cyclooxygenase cascade in the rabbit cornea.
Platelet-activating factor (PAF) is formed in the cornea after injury as well as by infiltrating inflammatory cells. We have studied the effects of PAF on the release and metabolism of arachidonic acid (AA) in the rabbit cornea. Corneal lipids were labeled in vivo by injecting [3H]AA and subsequently incubated in vitro with 100 nM PAF in the presence or absence of 10 microM BN50727, a PAF antagonist. The AA and eicosanoids released by incubated corneas were analyzed by high-performance liquid chromatography (HPLC). Tissue lipids were examined by mono- and bidimensional thin-layer chromatography (TLC). Within 5 min, PAF stimulated AA release to 76% above control levels. BN50727 inhibited the AA release elicited by PAF at all time points studied. The decreased content of [3H]AA in phosphatidylcholine (PC), phosphatidylethanolamine (PE) and phosphatidylinositol (PI) following PAF exposure and the lack of stimulation by PAF on the release of [3H] linoleic acid suggest that the cytosolic phospholipase A2 was activated. PAF also stimulated depletion of AA from the inositol lipids, phosphatidylinositol-4-phosphate (PIP) and phosphatidylinositol-4,5-biphosphate (PIP2) and increased content of [3H]AA into diacylglycerol (DAG) and phosphatidic acid (PA). This reaction indicates that PAF could also mediate activation of other phospholipases in the cornea. In addition, PAF preferentially stimulated the cyclooxygenase pathway. The PAF antagonist BN50727 mainly suppressed the PAF-stimulated release of PGE2. The antagonist did not inhibit lipoxygenase activity even after 30 min of PAF stimulation. These results suggest that PAF activate a phospholipase A2/cyclooxygenase pathway in the cornea via a PAF-receptor mechanism.(ABSTRACT TRUNCATED AT 250 WORDS)
Hurst
J S
JS
Department of Ophthalmology, Louisiana State University Eye Center, New Orleans 70112, USA.
Bazan
H E
HE
D000818:Animals; D016718:Arachidonic Acid / Q000096:biosynthesis; D001381:Azepines / Q000494:pharmacology; D003315:Cornea / Q000187:drug effects / Q000201:enzymology*; D015232:Dinoprostone / Q000096:biosynthesis; D005260:Female; D066298:In Vitro Techniques; D008297:Male; D010741:Phospholipases A / Q000378:metabolism*; D054467:Phospholipases A2; D010972:Platelet Activating Factor / Q000037:antagonists & inhibitors / Q000494:pharmacology*; D011451:Prostaglandin-Endoperoxide Synthases / Q000378:metabolism*; D011817:Rabbits; D058924:Thienopyridines; D014230:Triazoles / Q000494:pharmacology
D001381:Azepines; D010972:Platelet Activating Factor; D058924:Thienopyridines; D014230:Triazoles; D016718:Arachidonic Acid; D011451:Prostaglandin-Endoperoxide Synthases; D010741:Phospholipases A; D054467:Phospholipases A2; D015232:Dinoprostone; C067429:Setipafant