PAF-induced synthesis of tetraenoic and pentaenoic leukotrienes in the isolated rabbit lung.
In an isolated rabbit lung model, we tested the hypothesis that platelet-activating factor (PAF)-induced leukotriene (LT) synthesis is critically dependent on the free precursor fatty acid supply and the possible substitution of arachidonic acid (AA) by eicosapentaenoic acid (EPA). To augment the intravascular polymorphonuclear neutrophils (PMNs) in the isolated lung, human PMNs were infused into the pulmonary artery. LTs and hydroxyeicosatetra(penta)enoic acids were quantified with HPLC techniques. Application of PAF (5 microM) or AA (10 microM) provoked the generation of limited quantities of 4-series LTs and 5-hydroxyeicosatetraenoic acid (total sum of 5-lipoxygenase products approximately 7 and approximately 27 pmol/ml in lungs both with and without infused PMNs, respectively). Combined administration amplified 5-lipoxygenase product formation, with a predominance of cysteinyl-LT synthesis in lungs both without (total sum approximately 67 pmol/ml) and, much more strikingly, with (total sum approximately 308 pmol/ml) an infusion of neutrophils. EPA (10 microM) elicited exclusive generation of 5-series LTs and 5-hydroxyeicosapentaenoic acid (total sum approximately 82 pmol/ml). Dual stimulation with PAF and EPA provoked amplification of EPA-derived 5-lipoxygenase product formation, again with predominance of cysteinyl-LTs in lungs without (total sum approximately 224 pmol/ml) and, in particular, with (total sum approximately 545 pmol/ml) preceding microvascular PMN entrapment. Combined application of PAF, AA, and EPA resulted in the synthesis of LTs derived from both fatty acids, with a predominance of 5-series products. We conclude that the PAF-evoked 5-lipoxygenase product formation in the neutrophil-harboring lung capillary bed is critically dependent on intravascular precursor fatty acid supply, with EPA representing the preferred substrate compared with AA. PMN-related transcellular eicosanoid synthesis is suggested to underlie the predominant generation of cysteinyl-LTs. The supply of n-3 versus n-6 precursor fatty acid may thus have a major impact on inflammatory mediator generation.
Grimminger
F
F
Department of Internal Medicine, Justus-Liebig-University, D-35392 Giessen, Germany.
Mayer
K
K
Kiss
L
L
Walmrath
D
D
Seeger
W
W
D000818:Animals; D001094:Arachidonate 5-Lipoxygenase / Q000378:metabolism; D016718:Arachidonic Acid / Q000494:pharmacology; D002196:Capillaries / Q000378:metabolism; D003545:Cysteine / Q000096:biosynthesis; D004338:Drug Combinations; D015118:Eicosapentaenoic Acid / Q000494:pharmacology; D006801:Humans; D006893:Hydroxyeicosatetraenoic Acids / Q000096:biosynthesis / Q000378:metabolism; D066298:In Vitro Techniques; D015289:Leukotrienes / Q000096:biosynthesis*; D008168:Lung / Q000378:metabolism*; D009504:Neutrophils / Q000502:physiology; D010972:Platelet Activating Factor / Q000494:pharmacology*; D011652:Pulmonary Circulation / Q000502:physiology; D011817:Rabbits
D004338:Drug Combinations; D006893:Hydroxyeicosatetraenoic Acids; D015289:Leukotrienes; D010972:Platelet Activating Factor; C112381:cysteinyl-leukotriene; D016718:Arachidonic Acid; C022022:5-hydroxy-6,8,11,14-eicosatetraenoic acid; D015118:Eicosapentaenoic Acid; D001094:Arachidonate 5-Lipoxygenase; D003545:Cysteine