Endothelial barrier dysfunction and p42 oxidation induced by TNF-alpha are mediated by nitric oxide

The American Journal of Physiology
T J FerroA Johnson

Abstract

We tested the hypothesis that nitric oxide (.NO) mediates tumor necrosis factor-alpha (TNF-alpha)-induced alterations in permeability and actin of pulmonary artery endothelial monolayers (PAEM). The permeability of PAEM was assessed by the clearance rate of albumin labeled with Evans blue dye. The PAEM Triton-soluble ("cytoskeletal-nonassociated") and -insoluble ("cytoskeletal-associated") lysates were analyzed by Western blot for actin and oxidized protein using polyclonal antibodies to the COOH terminus of actin and dinitrophenylhydrazone (DNP), respectively. PAEM were incubated with TNF-alpha (100 U/ml) for 4 h. Incubation of PAEM with TNF-alpha resulted in increases in 1) the .NO oxidation product nitrite (NO2-), 2) nitrotyrosine immunofluorescence, 3) the oxidation of p42 (tentatively identified as actin), and 4) permeability to Evans blue dye-albumin. The .NO synthase inhibitor aminoguanidine (100 microM) prevented the TNF-alpha-induced increase in NO2-, nitrotyrosine immunofluorescence, oxidized p42, and permeability. Coincubation with L-arginine (200 microM) or the .NO mimic spermine-NO (1 microM) prevented the ablation of the response to TNF-alpha by aminoguanidine. The data indicate that TNF-alpha-induced increases in...Continue Reading

Citations

Feb 8, 2003·American Journal of Physiology. Lung Cellular and Molecular Physiology·Alma Siflinger-Birnboim, Arnold Johnson
Dec 24, 2005·Physiological Reviews·Dolly Mehta, Asrar B Malik
Oct 14, 2003·American Journal of Physiology. Lung Cellular and Molecular Physiology·Paul NeumannArnold Johnson
May 16, 2001·American Journal of Physiology. Gastrointestinal and Liver Physiology·A BananA Keshavarzian
Mar 11, 2000·American Journal of Physiology. Lung Cellular and Molecular Physiology·B A JohnsonP Davies
Jun 17, 2003·American Journal of Physiology. Lung Cellular and Molecular Physiology·Nancy GertzbergArnold Johnson
May 15, 2001·American Journal of Physiology. Lung Cellular and Molecular Physiology·I PetracheJ G Garcia
Dec 6, 2003·American Journal of Physiology. Lung Cellular and Molecular Physiology·Randall S Frey, Asrar B Malik
Jun 3, 2000·American Journal of Physiology. Lung Cellular and Molecular Physiology·T FerroA Johnson
Mar 26, 2003·American Journal of Respiratory Cell and Molecular Biology·Philip J O'ReillySadis Matalon
Jul 8, 2003·Journal of Applied Physiology·Fiemu E NwariakuLance S Terada
Nov 15, 2005·American Journal of Physiology. Lung Cellular and Molecular Physiology·Paul NeumannArnold Johnson
Sep 4, 2007·American Journal of Physiology. Lung Cellular and Molecular Physiology·Nancy GertzbergArnold Johnson
Apr 6, 2001·American Journal of Physiology. Lung Cellular and Molecular Physiology·K BoveA Johnson

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