Pretreatment of Human Cerebrovascular Endothelial Cells with CO‐releasing Molecule‐3 Interferes with JNK/AP‐1 Signaling and Suppresses LPS‐induced Proadhesive Phenotype. (January 2015)

• Record Type: Journal Article
• Authors: Serizawa, Fukashi; Patterson, Eric; Potter, Richard F.; Fraser, Douglas D.; Cepinskas, Gediminas
• Is Part Of: Microcirculation. Volume 22:Number 1(2015:Jan.)
• Page Start: 28
• Page End: 36
• Publication Date: 2015-01
• Subjects: Biological transport -- Periodicals; Microcirculation -- Physiology -- Periodicals; 612.135
• DOI: 10.1111/micc.12161
• Languages: English
• ISSNs: 1073-9688

Abstract

<abstract abstract-type="main" id="micc12161-abs-0001"> <title>Abstract</title> <sec id="micc12161-sec-0001" sec-type="section"> <title>Objective</title> <p>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.</p> </sec> <sec id="micc12161-sec-0002" sec-type="section"> <title>Methods</title> <p>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 <italic>in vitro</italic> model of endotoxemia. To this end, hCMEC/D3 were stimulated with LPS (1 <italic>μ</italic>g/mL) for six hours. In some experiments hCMEC/D3 were pretreated with CORM‐3 (200 <italic>μ</italic>mol/L) before LPS‐stimulation. PMN rolling/adhesion to hCMEC/D3 were assessed under conditions of laminar shear stress (0.7 dyn/cm<sup>2</sup>). In parallel, expression of adhesion molecules E‐selectin, ICAM‐1, and VCAM‐1 (qPCR), activation of transcription factors, NF‐<italic>κ</italic>B and AP‐1 (ELISA), and MAPK‐signaling (expression/phosphorylation of p38, ERK1/2, and JNK1/2; western blot) were assessed.</p> </sec> <sec id="micc12161-sec-0003" sec-type="section"> <title>Results</title> <p>The obtained results indicate that CORM‐3 pretreatment reduces PMN rolling/adhesion to LPS‐stimulated hCMEC/D3 (<italic>p</italic> < 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 (<italic>p</italic> < 0.05).</p> </sec> <sec id="micc12161-sec-0004" sec-type="section"> <title>Conclusions</title> <p>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.</p> </sec> </abstract>