title: Pretreatment of human cerebrovascular endothelial cells with CO-releasing molecule-3 interferes with JNK/AP-1 signaling and suppresses LPS-induced proadhesive phenotype.
abstract: OBJECTIVE
  Exogenously administered CO interferes with PMN recruitment to the inflamed organs. The mechanisms of CO-dependent modulation of vascular proadhesive phenotype, a key step in PMN recruitment, are unclear.
  
  
  METHODS
  We assessed the effects/mechanisms of CO liberated from a water-soluble CORM-3 on modulation of the proadhesive phenotype in hCMEC/D3 in an in vitro model of endotoxemia. To this end, hCMEC/D3 were stimulated with LPS (1 μg/mL) for six hours. In some experiments hCMEC/D3 were pretreated with CORM-3 (200 μmol/L) before LPS-stimulation. PMN rolling/adhesion to hCMEC/D3 were assessed under conditions of laminar shear stress (0.7 dyn/cm(2) ). In parallel, expression of adhesion molecules E-selectin, ICAM-1, and VCAM-1 (qPCR), activation of transcription factors, NF-κB and AP-1 (ELISA), and MAPK-signaling (expression/phosphorylation of p38, ERK1/2, and JNK1/2; western blot) were assessed.
  
  
  RESULTS
  The obtained results indicate that CORM-3 pretreatment reduces PMN rolling/adhesion to LPS-stimulated hCMEC/D3 (p < 0.05). Decreased PMN rolling/adhesion to hCMEC/D3 was associated with CORM-3-dependent inhibition of MAPK JNK1/2 activation (Tyr-phosphorylation), inhibition of transcription factor, AP-1 (c-Jun phosphorylation), and subsequent suppression of VCAM-1 expression (p < 0.05).
  
  
  CONCLUSIONS
  These findings indicate that CORM-3 pretreatment interferes with JNK/AP-1 signaling and suppresses LPS-induced upregulation of the proadhesive phenotype in hCMEC/D3.
authors.affiliation: Centre for Critical Illness Research, Lawson Health Research Institute, London, Ontario, Canada.
authors.forename: Fukashi
authors.initials: F
authors.lastname: Serizawa
authors.forename: Eric
authors.initials: E
authors.lastname: Patterson
authors.forename: Richard F
authors.initials: RF
authors.lastname: Potter
authors.forename: Douglas D
authors.initials: DD
authors.lastname: Fraser
authors.forename: Gediminas
authors.initials: G
authors.lastname: Cepinskas
mesh_terms: D000963:Antimetabolites / Q000494:pharmacology; D002248:Carbon Monoxide / Q000494:pharmacology; D002448:Cell Adhesion / Q000187:drug effects; D002460:Cell Line; D002560:Cerebrovascular Circulation / Q000187:drug effects; D042783:Endothelial Cells / Q000378:metabolism* / Q000473:pathology; D019446:Endotoxemia / Q000139:chemically induced / Q000378:metabolism* / Q000473:pathology; D006801:Humans; D018799:Intercellular Adhesion Molecule-1 / Q000096:biosynthesis; D008070:Lipopolysaccharides / Q000633:toxicity*; D048670:MAP Kinase Kinase 4 / Q000378:metabolism*; D020935:MAP Kinase Signaling System / Q000187:drug effects*; D009942:Organometallic Compounds / Q000494:pharmacology*; D018808:Transcription Factor AP-1 / Q000378:metabolism*; D019010:Vascular Cell Adhesion Molecule-1 / Q000096:biosynthesis
chemical_list: D000963:Antimetabolites; C573420:ICAM1 protein, human; D008070:Lipopolysaccharides; D009942:Organometallic Compounds; D018808:Transcription Factor AP-1; D019010:Vascular Cell Adhesion Molecule-1; C476530:tricarbonylchloro(glycinato)ruthenium(II); D018799:Intercellular Adhesion Molecule-1; D002248:Carbon Monoxide; D048670:MAP Kinase Kinase 4
keywords: CORM; E-selectin; ICAM-1; VCAM-1; adhesion molecules; anti-inflammatory; brain circulation; carbon monoxide; sepsis; transcription factors