title: Superoxide anion mediates the L-selectin down-regulation induced by non-steroidal anti-inflammatory drugs in human neutrophils.
abstract: Non-steroidal anti-inflammatory drugs (NSAIDs) induce the shedding of L-selectin in human neutrophils through a mechanism still not well understood. In this work we studied both the functional effect of NSAIDs on the neutrophils/endothelial cells dynamic interaction, and the potential involvement of reactive oxygen species (ROS) in the NSAIDs-mediated down-regulation of L-selectin. When human neutrophils were incubated with diclofenac, a significant reduction in the number of cells that rolled on activated endothelial cells was observed. Different NSAIDs (flufenamic acid, meclofenamic acid, diclofenac, indomethacin, nimesulide, flurbiprofen, meloxicam, phenylbutazone, piroxicam, ketoprofen and aspirin) caused variable increase in neutrophil intracellular ROS concentration, which was inversely proportional to the change produced in L-selectin surface expression. Pre-incubation of neutrophils with superoxide dismutase, but not with catalase, showed both a significant protective effect on the L-selectin down-regulation induced by several NSAIDs and a diminished effect of diclofenac on neutrophil rolling. Interestingly, diclofenac and flufenamic acid but not piroxicam significantly increased the extracellular superoxide anion production by neutrophils, and inhibition of nicotinamide adenine dinucleotide phosphate (NADPH)-oxidase activity with diphenyleneiodonium prevented the down-regulation of L-selectin by diclofenac. In accordance with these results, neutrophils from patients with chronic granulomatous disease, a hereditary disease in which neutrophils show a reduced capacity to form superoxide radicals, exhibited a lower down-regulation of L-selectin (IC50: 15.3 μg/ml) compared to normal controls (IC50: 5.6 μg/ml) in response to diclofenac.
  
  
  CONCLUSIONS
  A group of NSAIDs is capable of interfering with the ability of neutrophils to interact with endothelial cells by triggering L-selectin-shedding through the NADPH-oxidase-dependent generation of superoxide anion at the plasma membrane.
authors.affiliation: Servicio de Reumatología, Hospital Universitario de Canarias, La Laguna, Spain.
authors.forename: Maria
authors.initials: M
authors.lastname: Domínguez-Luis
authors.forename: Ada
authors.initials: A
authors.lastname: Herrera-García
authors.forename: Maria
authors.initials: M
authors.lastname: Arce-Franco
authors.forename: Estefania
authors.initials: E
authors.lastname: Armas-González
authors.forename: Marta
authors.initials: M
authors.lastname: Rodríguez-Pardo
authors.forename: Fabian
authors.initials: F
authors.lastname: Lorenzo-Díaz
authors.forename: Manuel
authors.initials: M
authors.lastname: Feria
authors.forename: Susana
authors.initials: S
authors.lastname: Cadenas
authors.forename: Francisco
authors.initials: F
authors.lastname: Sánchez-Madrid
authors.forename: Federico
authors.initials: F
authors.lastname: Díaz-González
mesh_terms: D051722:ADAM Proteins / Q000235:genetics / Q000378:metabolism; D000072198:ADAM17 Protein; D000293:Adolescent; D000818:Animals; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000494:pharmacology*; D002450:Cell Communication / Q000187:drug effects; D002461:Cell Line, Transformed; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D002465:Cell Movement / Q000187:drug effects; D002478:Cells, Cultured; D002648:Child; D004008:Diclofenac / Q000494:pharmacology*; D015536:Down-Regulation / Q000187:drug effects*; D005439:Flufenamic Acid / Q000494:pharmacology*; D006105:Granulomatous Disease, Chronic / Q000235:genetics / Q000276:immunology / Q000378:metabolism / Q000473:pathology; D061307:Human Umbilical Vein Endothelial Cells / Q000166:cytology / Q000187:drug effects / Q000276:immunology / Q000378:metabolism; D006801:Humans; D019041:L-Selectin / Q000378:metabolism*; D008297:Male; D051379:Mice; D008817:Mice, Mutant Strains; D019255:NADPH Oxidases / Q000235:genetics / Q000378:metabolism; D009504:Neutrophils / Q000187:drug effects* / Q000276:immunology / Q000378:metabolism / Q000473:pathology; D013481:Superoxides / Q000378:metabolism*
chemical_list: D000894:Anti-Inflammatory Agents, Non-Steroidal; C546935:SELL protein, human; D013481:Superoxides; D019041:L-Selectin; D004008:Diclofenac; D005439:Flufenamic Acid; D019255:NADPH Oxidases; C060505:neutrophil cytosolic factor 1; D051722:ADAM Proteins; D000072198:ADAM17 Protein