Intrinsic 5-lipoxygenase activity is required for neutrophil responsivity.
We found that intrinsic neutrophil 5-lipoxygenase activity was necessary for human neutrophil adherence and chemotaxis in vitro and human neutrophil-mediated acute edematous injury in isolated perfused rat lungs given interleukin 8 intratracheally. Treatment with either Zileuton (a specific reversible competitive inhibitor of 5-lipoxygenase) or MK886 (a specific irreversible inhibitor of the 5-lipoxygenase activator protein) prevented stimulated neutrophil adherence and chemotaxis (but not superoxide anion production) in vitro. Zileuton- or MK886-inhibited neutrophil chemotaxis was not restored by adding leukotriene B4 in vitro. Perfusion with neutrophils and either Zileuton or MK886, or with MK886-pretreated neutrophils (without adding MK886 to the perfusate), also prevented lung injury (reflected by lung weight gain and lung Ficoll retention) and perfusate leukotriene B4 increases in isolated rat lungs given interleukin 8 intratracheally. Again, adding leukotriene B4 to the perfusate did not damage interleukin 8-treated isolated lungs perfused with Zileuton-inhibited neutrophils. We conclude that intrinsic 5-lipoxygenase activity is required for neutrophil adherence and chemotaxis and neutrophil-mediated lung injury.
Guidot
D M
DM
Webb-Waring Institute for Biomedical Research, University of Colorado Health Sciences Center, Denver 80262.
Repine
M J
MJ
Westcott
J Y
JY
Repine
J E
JE
D000818:Animals; D001094:Arachidonate 5-Lipoxygenase / Q000097:blood*; D002448:Cell Adhesion / Q000187:drug effects; D002634:Chemotaxis, Leukocyte / Q000187:drug effects; D003574:Cytochrome c Group / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D006801:Humans; D006918:Hydroxyurea / Q000031:analogs & derivatives / Q000494:pharmacology; D066298:In Vitro Techniques; D007211:Indoles / Q000494:pharmacology; D016209:Interleukin-8 / Q000494:pharmacology*; D007700:Kinetics; D020024:Leukotriene Antagonists; D007975:Leukotriene B4 / Q000378:metabolism / Q000494:pharmacology; D016859:Lipoxygenase Inhibitors; D008168:Lung / Q000187:drug effects / Q000378:metabolism*; D008297:Male; D009504:Neutrophils / Q000187:drug effects / Q000201:enzymology / Q000502:physiology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D011994:Recombinant Proteins / Q000494:pharmacology; D013481:Superoxides / Q000378:metabolism; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D002722:Chlorobenzenes
D001094:Arachidonate 5-Lipoxygenase; D003574:Cytochrome c Group; D006918:Hydroxyurea; D007211:Indoles; D016209:Interleukin-8; D020024:Leukotriene Antagonists; D007975:Leukotriene B4; D016859:Lipoxygenase Inhibitors; D011994:Recombinant Proteins; D013481:Superoxides; D013755:Tetradecanoylphorbol Acetate; C060893:MK-886; D002722:Chlorobenzenes; C063449:zileuton