Carbon Monoxide-Releasing Molecule-401 Suppresses Polymorphonuclear Leukocyte Migratory Potential by Modulating F-Actin Dynamics.
Carbon monoxide-releasing molecules (CORMs) suppress inflammation by reducing polymorphonuclear leukocyte (PMN) recruitment to the affected organs. We investigated modulation of PMN-endothelial cell adhesive interactions by water-soluble CORM-401 using an experimental model of endotoxemia in vitro. Human umbilical vein endothelial cells (HUVEC) grown on laminar-flow perfusion channels were stimulated with 1 μg/mL lipopolysaccharide for 6 hours and perfused with 100 μmol/L CORM-401 (or inactive compound iCORM-401)-pretreated PMN for 5 minutes in the presence of 1.0 dyn/cm2shear stress.
PMN co-cultures were perfused for additional 15 minutes with PMN-free medium containing CORM-401/inactive CORM-401. The experiments were videorecorded (phase-contrast microscopy), and PMN adhesion/migration were assessed off-line. In parallel, CORM-401-dependent modulation of PMN chemotaxis, F-actin expression/distribution, and actin-regulating pathways [eg, p21-activated protein kinases (PAK1/2) and extracellular signal-regulated kinase (ERK)/C-Jun N-terminal kinase (JNK) mitogen-activated protein kinases (MAPK)] were assessed in response to N-formyl-methionyl-leucyl-phenylalanine (fMLP) stimulation. Pretreating PMN with CORM-401 did not suppress PMN adhesion to HUVEC, but significantly reduced PMN transendothelial migration (P < 0.0001) and fMLP-induced PMN chemotaxis (ie, migration directionality and velocity). These changes were associated with CORM-401-dependent suppression of F-actin levels/cellular distribution and fMLP-induced phosphorylation of PAK1/2 and ERK/JNK MAPK (P < 0.05). CORM-401 had no effect on p38 MAPK activation. In summary, this study demonstrates, for the first time, CORM-401-dependent suppression of neutrophil migratory potential associated with modulation of PAK1/2 and ERK/JNK MAPK signaling and F-actin dynamics.
Centre for Critical Illness Research, Lawson Health Research Institute, London, Ontario, Canada.
Ken
K
Inoue
Centre for Critical Illness Research, Lawson Health Research Institute, London, Ontario, Canada.
Eric K
EK
Patterson
Department of Chemistry, McMaster University, Hamilton, Ontario, Canada.
Alfredo
A
Capretta
Centre for Critical Illness Research, Lawson Health Research Institute, London, Ontario, Canada; Department of Medical Biophysics, University of Western Ontario, London, Ontario, Canada.
Abdel R
AR
Lawendy
Centre for Critical Illness Research, Lawson Health Research Institute, London, Ontario, Canada; Department of Paediatrics, University of Western Ontario, London, Ontario, Canada.
Douglas D
DD
Fraser
Centre for Critical Illness Research, Lawson Health Research Institute, London, Ontario, Canada; Department of Medical Biophysics, University of Western Ontario, London, Ontario, Canada. Electronic address: gcepinsk@uwo.ca.
Gediminas
G
Cepinskas
D000199:Actins / Q000378:metabolism; D018821:CD18 Antigens / Q000378:metabolism; D002248:Carbon Monoxide / Q000378:metabolism*; D002448:Cell Adhesion / Q000502:physiology; D002465:Cell Movement / Q000502:physiology*; D002478:Cells, Cultured; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D048670:MAP Kinase Kinase 4 / Q000378:metabolism; D020935:MAP Kinase Signaling System / Q000502:physiology; D020929:Mitogen-Activated Protein Kinase Kinases / Q000378:metabolism; D009504:Neutrophils / Q000502:physiology*; D010766:Phosphorylation / Q000502:physiology; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000378:metabolism
D000199:Actins; D018821:CD18 Antigens; D019408:Platelet Endothelial Cell Adhesion Molecule-1; D002248:Carbon Monoxide; D048670:MAP Kinase Kinase 4; D020929:Mitogen-Activated Protein Kinase Kinases