The sensitivity of bovine corneal epithelial lyso-PAF acetyltransferase to cyclooxygenase and lipoxygenase inhibitors is independent of arachidonate metabolites.
Lyso-platelet-activating factor (PAF) acetyltransferase is a critical regulatory step in PAF synthesis. PAF accumulates in the cornea in response to injury and is a potent inflammatory mediator that stimulates corneal cyclooxygenase but not lipoxygenase reactions. 12(S)-hydroxyeicosatetraenoic (HETE) acid, a major lipoxygenase product of mammalian corneas, is also generated during injury. In the bovine corneal epithelium, both PAF acetyltransferase and 12-lipoxygenase are microsomal enzymes. A potential interaction between these two lipid mediators was, therefore, examined. PAF acetyltransferase activity was assayed by determining radioactivity in the trichloroacetic acid-precipitated complex of [3H]PAF bound to albumin, formed after incubation of corneal epithelial microsomes with lyso-PAF and [3H]acetyl CoA. Lipoxygenase metabolism by bovine corneal epithelial microsomes was also studied. While 12(S)-HETE did not activate lyso-PAF acetyltransferase, PAF synthesis was decreased when microsomes were treated with lipoxygenase inhibitors. The IC50 values for nordihydroguaiaretic acid (NDGA), and baicalein were 75 and 105 microM, respectively. The IC50 value for CDC, a more potent inhibitor of platelet 12-lipoxygenase, was greater than 200 microM. The results indicate that concentrations suppressing lyso-PAF acetyltransferase activity exceed those required to inhibit lipoxygenases from bovine corneal epithelia. Similarly, concentrations of aspirin and indomethacin, cyclooxygenase inhibitors that decrease PAF formation, were greater than those reported to block prostaglandin generation. A number of other compounds, some structurally similar to the lipoxygenase inhibitors that suppress lyso-PAF acetyltransferase, and others unrelated chemically but known as anti-oxidant and cationic-chelators, also inhibited lyso-PAF acetyltransferase. This suggests that lyso-PAF acetyltransferase activity is inhibited by mechanisms independent of lipoxygenase and cyclooxygenase metabolites.
Hurst
J S
JS
Department of Ophthalmology, Louisiana State University Eye Center, New Orleans, USA.
Bazan
H E
HE
D019377:12-Hydroxy-5,8,10,14-eicosatetraenoic Acid / Q000378:metabolism; D000123:Acetyltransferases / Q000187:drug effects*; D000818:Animals; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000494:pharmacology; D000975:Antioxidants / Q000494:pharmacology; D016718:Arachidonic Acid / Q000378:metabolism*; D002417:Cattle; D016861:Cyclooxygenase Inhibitors / Q000494:pharmacology*; D019573:Epithelium, Corneal / Q000187:drug effects* / Q000201:enzymology; D016859:Lipoxygenase Inhibitors / Q000494:pharmacology*
D000894:Anti-Inflammatory Agents, Non-Steroidal; D000975:Antioxidants; D016861:Cyclooxygenase Inhibitors; D016859:Lipoxygenase Inhibitors; D016718:Arachidonic Acid; D019377:12-Hydroxy-5,8,10,14-eicosatetraenoic Acid; D000123:Acetyltransferases; C046035:1-alkylglycerophosphocholine acetyltransferase