• Fukashi Serizawa, Eric Patterson, Richard F Potter, Douglas D Fraser, and Gediminas Cepinskas.
• Centre for Critical Illness Research, Lawson Health Research Institute, London, Ontario, Canada.
• Microcirculation. 2015 Jan 1;22(1):28-36.

ObjectiveExogenously 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.MethodsWe 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.ResultsThe 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).ConclusionsThese findings indicate that CORM-3 pretreatment interferes with JNK/AP-1 signaling and suppresses LPS-induced upregulation of the proadhesive phenotype in hCMEC/D3.© 2014 John Wiley & Sons Ltd.

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