Faculty Collaboration Database

Modulation of endotoxin-induced endothelial function by calcium/calmodulin-dependent protein kinase. Shock 2003 Aug;20(2):176-82

Date

07/17/2003

Pubmed ID

12865664

DOI

10.1097/01.shk.0000074789.29800.a5

Scopus ID

2-s2.0-1542541983 (requires institutional sign-in at Scopus site)   18 Citations

Abstract

Endothelial cells facilitate sepsis-induced neutrophil adherence through the production of adhesion molecules and proinflammatory cytokines. The production of these factors requires coordinated intracellular inflammatory signaling. Recently, patients prone to sepsis-induced complications have been shown to have derangements in intracellular calcium and potentially calcium/calmodulin-dependent protein kinase (CaMK) activity, but the impact of these impairments is unknown. Human umbilical vein endothelial vein endothelial cells (HUVECs) were exposed to lipopolysaccharide (LPS) for various periods of time. Select HUVECs were pretreated with an inhibitor of CaMK II, KN62. Total cellular and nuclear proteins were extracted and analyzed for various components of the Toll-mediated signal cascade. Neutrophil adhesion was assayed fluorometrically using calcein-labeled neutrophils on treated HUVECs. LPS stimulation led to mitogen-activated protein kinase activation and translocation of activator protein-1 (AP-1) and nuclear factor (NF)-kappaB. CaMK blockade inhibited LPS induced ERK 1/2 and JNK but enhanced p38 activity. This selective MAPK inhibition was associated with a reduction in AP-1 activity, with no affect on NF-kappaB activity. Associated with this altered cell signaling was increased ICAM-1 production and enhanced neutrophil adhesion. Altered CaMK activity resulted in dysregulated mitogen-activated protein kinase signaling, demonstrated by reduced ERK 1/2 and JNK activity but enhanced p38 activity. This altered signaling is associated with reduced AP-1 activation and unaffected NF-kappaB activation. Neutrophil adhesion, however, is enhanced presumably through increased ICAM-1 production. Therefore, CaMK inhibition of endothelial cells, characteristic of sustained increases in intracellular calcium, appears to result in a dysregulated proadhesive phenotype.

Author List

Cuschieri J, Gourlay D, Garcia I, Jelacic S, Maier RV

Author

David M. Gourlay MD Chief, Professor in the Surgery department at Medical College of Wisconsin

MESH terms used to index this publication - Major topics in bold

Blotting, Western
Calcium
Calcium-Calmodulin-Dependent Protein Kinases
Cell Adhesion
Cell Differentiation
Cell Nucleus
Cell Survival
Cells, Cultured
DNA
Endothelium, Vascular
Endotoxins
Escherichia coli
Fluoresceins
Humans
Intercellular Adhesion Molecule-1
Lipopolysaccharides
MAP Kinase Signaling System
Mitogen-Activated Protein Kinase 1
Mitogen-Activated Protein Kinase 3
Mitogen-Activated Protein Kinases
Neutrophils
Phenotype
Sepsis
Time Factors
Transcription Factor AP-1
Umbilical Veins
p38 Mitogen-Activated Protein Kinases