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Am J Physiol Lung Cell Mol Physiol 280: L1168-L1178, 2001;
1040-0605/01 $5.00
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Vol. 280, Issue 6, L1168-L1178, June 2001

Differential effect of MLC kinase in TNF-alpha -induced endothelial cell apoptosis and barrier dysfunction

Irina Petrache1, Alexander D. Verin1, Michael T. Crow2, Anna Birukova1, Feng Liu1, and Joe G. N. Garcia1

1 Division of Pulmonary and Critical Care Medicine, Department of Medicine, Johns Hopkins University School of Medicine, and 2 Vascular Biology Unit, Laboratory of Cardiovascular Science, Gerontology Research Center, National Institute on Aging, National Institutes of Health, Baltimore, Maryland 21224

Tumor necrosis factor (TNF)-alpha is released in acute inflammatory lung syndromes linked to the extensive vascular dysfunction associated with increased permeability and endothelial cell apoptosis. TNF-alpha induced significant decreases in transcellular electrical resistance across pulmonary endothelial cell monolayers, reflecting vascular barrier dysfunction (beginning at 4 h and persisting for 48 h). TNF-alpha also triggered endothelial cell apoptosis beginning at 4 h, which was attenuated by the caspase inhibitor Z-Val-Ala-Asp-fluoromethylketone. Exploring the involvement of the actomyosin cytoskeleton in these important endothelial cell responses, we determined that TNF-alpha significantly increased myosin light chain (MLC) phosphorylation, with prominent stress fiber and paracellular gap formation, which paralleled the onset of decreases in transcellular electrical resistance and enhanced apoptosis. Reductions in MLC phosphorylation by the inhibition of either MLC kinase (ML-7, cholera toxin) or Rho kinase (Y-27632) dramatically attenuated TNF-alpha -induced stress fiber formation, indexes of apoptosis, and caspase-8 activity but not TNF-alpha -induced barrier dysfunction. These studies indicate a central role for the endothelial cell cytoskeleton in TNF-alpha -mediated apoptosis, whereas TNF-alpha -induced vascular permeability appears to evolve independently of contractile tension generation.

caspases; cytoskeleton; acute lung injury; permeability; Rho kinase; tumor necrosis factor-alpha


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