Release of a neutrophil-derived vasoconstrictor agent which augments platelet-induced contractions of blood vessels in vitro.
1. The effects of neutrophil-derived products on vascular tone in vitro were examined by adding purified rabbit neutrophils to siliconized organ baths containing rings of rabbit thoracic aorta. 2. Neutrophil-derived products induced a concentration-dependent contraction of the blood vessels generating 3.9 +/- 0.2 g of tension at a cell concentration of 2 X 10(6) ml-1. This contractile response was not dependent on an intact endothelium and was not ameliorated by treatment of the neutrophils with inhibitors of lipoxygenase or cyclo-oxygenase enzymes, by free radical scavengers or by the use of end organ antagonists to angiotensin II, desArg9-bradykinin, histamine, catecholamines or acetylcholine. 3. Neutrophils stimulated with phorbol myristate acetate, formyl methionyl-leucyl-phenylalanine or calcium ionophore A23187 released a contractile factor into the supernatant which produced qualitatively similar contractions compared to those elicited by incubation with intact unstimulated neutrophils. 4. Ten to twenty times more platelets were required to evoke equivalent contractions to those observed with neutrophils. However, neutrophil supernates significantly augmented platelet-mediated contractions (P less than 0.001). 5. Contractions elicited by neutrophils and supernates derived from activated neutrophils were partially antagonized by 5-hydroxytryptamine (5-HT) receptor antagonists, methysergide and ketanserin. However, the inhibition of exogenous 5-HT-induced contractions on rabbit aorta and rat stomach strips by both antagonists was greater than the inhibition of contractions produced by neutrophils and neutrophil-derived supernates. 6. Extraction of the biologically active material from supernatants of activated neutrophils into acetone, but not chloroform-methanol or ethyl acetate, suggests the contractile factor may be a protein/peptide. Partial purification on a Sephadex G100 column yields a contractile factor with a molecular weight of less than 4,000 daltons. 7. This factor may augment vascular tone, either directly or by interactions with platelets, in pathophysiological states associated with neutrophil activation and accumulation.
Sessa
W C
WC
Department of Pharmacology, New York Medical College, Valhalla 10595.
Mullane
K M
KM
D000818:Animals; D001013:Aorta, Thoracic / Q000187:drug effects; D001792:Blood Platelets / Q000502:physiology*; D000001:Calcimycin / Q000494:pharmacology; D066298:In Vitro Techniques; D016859:Lipoxygenase Inhibitors; D008297:Male; D008970:Molecular Weight; D009119:Muscle Contraction / Q000187:drug effects; D009129:Muscle Tonus / Q000187:drug effects; D009131:Muscle, Smooth, Vascular / Q000187:drug effects*; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000494:pharmacology; D009504:Neutrophils / Q000502:physiology*; D011817:Rabbits; D012701:Serotonin / Q000494:pharmacology; D012702:Serotonin Antagonists / Q000494:pharmacology; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014662:Vasoconstrictor Agents / Q000494:pharmacology*
D016859:Lipoxygenase Inhibitors; D012702:Serotonin Antagonists; D014662:Vasoconstrictor Agents; D012701:Serotonin; D000001:Calcimycin; D009240:N-Formylmethionine Leucyl-Phenylalanine; D013755:Tetradecanoylphorbol Acetate