Modulation of endotoxin-induced endothelial activity by microtubule depolymerization. J Trauma 2003 Jan;54(1):104-12; discussion 112-3
Date
01/25/2003Pubmed ID
12544905DOI
10.1097/00005373-200301000-00013Scopus ID
2-s2.0-0037237407 (requires institutional sign-in at Scopus site) 16 CitationsAbstract
BACKGROUND: Endotoxin not only activates the Toll-mediated signaling pathway within endothelial cells that leads to neutrophil migration but also causes the polymerization of microtubules. The potential role of this polymerization event, however, is unknown.
METHODS: Human umbilical vein endothelial cells stimulated with endotoxin were pretreated with or without the microtubule depolymerizing agent colchicine. Toll-mediated signaling events and protein production were in turn investigated by Western blot, gel shift, and enzyme-linked immunosorbent assay. Finally, neutrophil adhesion was assayed fluorometrically under the various conditions.
RESULTS: Endotoxin led to activation of the various Toll-mediated pathways, production of intercellular adhesion molecule-1 and interleukin-8, and subsequent neutrophil adhesion. Pretreatment with colchicine led to selective inhibition of anti-dual phosphorylated extracellular signal-regulated kinase-1/2, anti-dual phosphorylated c-jun N-terminal kinase, and adaptor protein-1; selective enhancement of p38; and no effect on nuclear factor-kappaB. This selective modulation of intracellular signaling resulted in attenuated intercellular adhesion molecule-1, interleukin-8 and prostaglandin E2 production, but enhanced cyclooxygenase-2 expression. As a result, microtubule disruption led to a significant reduction in neutrophil adhesion.
CONCLUSION: Microtubule formation is essential to optimal endotoxin-induced intracellular signaling through anti-dual phosphorylated extracellular signal-regulated kinase-1/2, anti-dual phosphorylated c-jun N-terminal kinase, and adaptor protein-1. Failure of these signaling events is associated with a marked reduction in the formation of a proadhesive phenotype that may prove to be beneficial in modulating neutrophil recruitment during sepsis.
Author List
Cuschieri J, Gourlay D, Garcia I, Jelacic S, Maier RVAuthor
David M. Gourlay MD Chief, Professor in the Surgery department at Medical College of WisconsinMESH terms used to index this publication - Major topics in bold
Adaptor Protein Complex 1Blotting, Western
Cells, Cultured
Colchicine
Cyclooxygenase 2
Dinoprostone
Drosophila Proteins
Electrophoretic Mobility Shift Assay
Endothelium, Vascular
Enzyme-Linked Immunosorbent Assay
Escherichia coli
Humans
Intercellular Adhesion Molecule-1
Interleukin-8
Isoenzymes
JNK Mitogen-Activated Protein Kinases
Lipopolysaccharides
Membrane Glycoproteins
Membrane Proteins
Microtubules
Mitogen-Activated Protein Kinase 3
Mitogen-Activated Protein Kinases
NF-kappa B
Neutrophil Activation
Neutrophil Infiltration
Polymers
Prostaglandin-Endoperoxide Synthases
Receptors, Cell Surface
Sepsis
Signal Transduction
Toll-Like Receptors
Umbilical Veins