title: FMLP causes eicosanoid-dependent vasoconstriction and edema in lungs from endotoxin-primed rats.
abstract: Recruitment of inflammatory cells to the lung capillaries has been proposed as an important step in the sequence of events that lead to acute lung injury. Frequently, in the clinical setting, bacteremia and sepsis syndrome precede the acute lung failure and endotoxin priming may represent a comparable paradigm, useful for experimental pursuit. Following addition of the chemotactic tripeptide FMLP (10(-9) to 10(-6) M) to the cell-free, salt solution perfusate of isolated rat lungs, only a small degree of vasoconstriction was observed. However, in lungs isolated from rats that received 2 mg/kg intraperitoneal Salmonella enteritidis endotoxin 2 h before lung perfusion, FMLP dose dependently caused a large, transient pulmonary pressor response, edema formation, and release of large amounts of thromboxane and leukotriene B4. Since in vitro priming with endotoxin, direct vascular injury by neutrophil elastase, nor direct stimulation with FMLP of pulmonary artery rings from endotoxin-pretreated rats, mimicked the effects of in vivo endotoxin priming, we conclude that the presence of inflammatory cells in the lung capillaries accounted for the large amount of eicosanoids produced by the lungs after FMLP stimulation. In fact, by retrograde lavage of the lung circulation with a collagenase solution, previously adherent cell clumps were mobilized and identified. These cell clumps, composed of red blood cells, neutrophils, and platelets, were not seen in the vascular lavage sediment obtained from unprimed control lungs. Indomethacin, a thromboxane antagonist, AA861, a 5-lipoxygenase inhibitor, and WEB 2086, a platelet-activating factor (PAF) antagonist, reduced the thromboxane synthesis and release after FMLP (10(-7) M) in in vivo endotoxin-primed lungs. None of the inhibitors employed exclusively inhibited only one particular eicosanoid mediator but rather affected the release of several mediators, suggesting a close link between the different synthetic arachidonic acid pathways. An inhibitor of phospholipase C (2-nitro-4-carboxyphenyl-N,N-diphenylcarbamate), NCDC, but not an inhibitor of phospholipase D (Wortmannin) or of protein kinase C (staurosporine) inhibited the FMLP-stimulated pulmonary pressure rise and eicosanoid release in endotoxin-primed lungs in vivo. Our data suggest that eicosanoids (in particular thromboxane) released from cells trapped in the lung circulation, but not from constitutive lung cells, contribute to vasoconstriction and edema formation caused by the chemoattractant FMLP in endotoxin-primed lungs.
authors.affiliation: Cardiovascular Pulmonary Research Laboratory, University of Colorado Health Sciences Center, Denver 80262.
authors.forename: N F
authors.initials: NF
authors.lastname: Voelkel
authors.forename: J
authors.initials: J
authors.lastname: Czartolomna
authors.forename: J
authors.initials: J
authors.lastname: Simpson
authors.forename: R C
authors.initials: RC
authors.lastname: Murphy
mesh_terms: D000470:Alkaloids / Q000494:pharmacology; D000730:Androstadienes / Q000494:pharmacology; D000818:Animals; D002219:Carbamates / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D015777:Eicosanoids / Q000032:analysis / Q000096:biosynthesis*; D004731:Endotoxins / Q000494:pharmacology*; D066298:In Vitro Techniques; D008297:Male; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000494:pharmacology*; D048448:Phenylcarbamates; D010739:Phospholipase D / Q000037:antagonists & inhibitors; D011493:Protein Kinase C / Q000037:antagonists & inhibitors; D011654:Pulmonary Edema / Q000139:chemically induced* / Q000503:physiopathology; D051381:Rats; D011919:Rats, Inbred Strains; D012477:Salmonella enteritidis; D019311:Staurosporine; D010738:Type C Phospholipases / Q000037:antagonists & inhibitors; D014661:Vasoconstriction / Q000187:drug effects* / Q000502:physiology; D000077191:Wortmannin
chemical_list: D000470:Alkaloids; D000730:Androstadienes; D002219:Carbamates; D015777:Eicosanoids; D004731:Endotoxins; D048448:Phenylcarbamates; C025508:2-nitro-4-carboxyphenyl-N,N-diphenylcarbamate; D009240:N-Formylmethionine Leucyl-Phenylalanine; D011493:Protein Kinase C; D010738:Type C Phospholipases; D010739:Phospholipase D; D019311:Staurosporine; D000077191:Wortmannin