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Carbon monoxide (CO)‐dependent modulation of polymorphonuclear leukocyte (PMN) adhesion to human cerebrovascular endothelial cells in an in vitro model of endotoxemia
Author(s) -
Serizawa Fukashi,
Patterson Eric,
Potter Richard F,
Fraser Duglas,
Cepinskas Gediminas
Publication year - 2013
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.27.1_supplement.57.10
Subject(s) - p38 mitogen activated protein kinases , cell adhesion molecule , adhesion , western blot , phosphorylation , chemistry , in vitro , inflammation , stimulation , e selectin , pharmacology , cell adhesion , immunology , medicine , mapk/erk pathway , biochemistry , organic chemistry , gene
CO liberated from a water‐soluble CO‐releasing molecule (CORM‐3) suppresses sepsis‐induced inflammation. However, the exact mechanisms are poorly understood. In this study we examined the mechanisms of CORM‐3‐dependent modulation of PMN adhesion in an in vitro model of endotoxemia employing human‐derived cerebrovascular endothelial cells, hCMEC/D3 (provided by Dr. P. Couraud, INSERM, Paris). hCMEC/D3 were stimulated with LPS (1μg/mL) for 6 hrs. In some experiments hCMEC/D3 were: 1) pre‐treated, 2) co‐treated, or 3) treated with CORM‐3 or inactive CORM‐3 (iCORM‐3) (200μM) before/during/following stimulation with LPS, respectively. PMN rolling/adhesion to hCMEC/D3 was assessed under conditions of flow (shear stress 0.7dyn/cm 2 ). In parallel, MAPK‐signaling (phosphorylation of p38, ERK1/2 and JNK1/2; Western Blot) and expression of adhesion molecules (E‐selectin, ICAM‐1 and VCAM‐1; RT‐PCR) were also assessed. CORM‐3 significantly reduced PMN adhesion to hCMEC/D3 with the most profound effects seen in “pre‐treatment” group. This was accompanied by decreased levels of JNK1/2 Tyr‐phosphorylation and suppressed expression of E‐selectin and VCAM‐1. The results indicate that CORM‐3 interferes with activation of JNK1/2 phosphorylation and expression of hCMEC/D3 pro‐adhesive phenotype, thus potentially could be used to reduce systemic inflammation (HSFO‐393, HSFO‐NA6914, IRF‐04–79)