HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) All Versions of this Article: 290/5/L919    most recent 00434.2005v1 Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in Web of Science Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Web of Science (9) Citing Articles via Google Scholar Google Scholar Articles by Moldobaeva, A. Articles by Pearse, D. B. Search for Related Content PubMed PubMed Citation Articles by Moldobaeva, A. Articles by Pearse, D. B.

Role of protein kinase G in barrier-protective effects of cGMP in human pulmonary artery endothelial cells

¹Division of Pulmonary and Critical Care Medicine, Department of Medicine, and ²Department of Pathology, The Johns Hopkins Medical Institutions, Baltimore, Maryland

Submitted 11 October 2005 ; accepted in final form 2 December 2005

Increases in endothelial cGMP prevent oxidant-mediated endothelial barrier dysfunction, but the downstream mechanisms remain unclear. To determine the role of cGMP-dependent protein kinase (PKG)_I, human pulmonary artery endothelial cells (HPAEC) lacking PKG_I expression were infected with a recombinant adenovirus encoding PKG_I (Ad.PKG) and compared with uninfected and control-infected (Ad.gal) HPAEC. Transendothelial electrical resistance (TER), an index of permeability, was measured after H₂O₂ (250 µM) exposure with or without pretreatment with 8-(4-chlorophenylthio)guanosine 3',5'-cyclic monophosphate (CPT-cGMP). HPAEC infected with Ad.PKG, but not Ad.gal, expressed PKG_I protein and demonstrated Ser²³⁹ and Ser¹⁵⁷ phosphorylation of vasodilator-stimulated phosphoprotein after treatment with CPT-cGMP. Adenoviral infection decreased basal permeability equally in Ad.PKG- and Ad.gal-infected HPAEC compared with uninfected cells. Treatment with CPT-cGMP (100 µM) caused a PKG_I-independent decrease in permeability (8.2 ± 0.6%). In all three groups, H₂O₂ (250 µM) caused a similar 35% increase in permeability associated with increased actin stress fiber formation, intercellular gaps, loss of membrane VE-cadherin, and increased intracellular Ca²⁺ concentration ([Ca²⁺]_i). In uninfected and Ad.gal-infected HPAEC, pretreatment with CPT-cGMP (100 µM) partially blocked the increased permeability induced by H₂O₂. In Ad.PKG-infected HPAEC, CPT-cGMP (50 µM) prevented the H₂O₂-induced TER decrease, cytoskeletal rearrangement, and loss of junctional VE-cadherin. CPT-cGMP attenuated the peak [Ca²⁺]_i caused by H₂O₂ similarly (23%) in Ad.gal- and Ad.PKG-infected HPAEC, indicating a PKG_I-independent effect. These data suggest that cGMP decreased HPAEC basal permeability by a PKG_I-independent process, whereas the ability of cGMP to prevent H₂O₂-induced barrier dysfunction was predominantly mediated by PKG_I through a Ca²⁺-independent mechanism.

cAMP; pulmonary edema; acute lung injury; hydrogen peroxide

This article has been cited by other articles:

				E. P. Schmidt, M. Damarla, O. Rentsendorj, L. E. Servinsky, B. Zhu, A. Moldobaeva, A. Gonzalez, P. M. Hassoun, and D. B. Pearse Soluble guanylyl cyclase contributes to ventilator-induced lung injury in mice Am J Physiol Lung Cell Mol Physiol, December 1, 2008; 295(6): L1056 - L1065. [Abstract] [Full Text] [PDF]

				J. M. Dodd-o, M. L. Hristopoulos, K. Kibler, J. Gutkowska, S. Mukaddam-Daher, A. Gonzalez, L. E. Welsh-Servinsky, and D. B. Pearse The role of natriuretic peptide receptor-A signaling in unilateral lung ischemia-reperfusion injury in the intact mouse Am J Physiol Lung Cell Mol Physiol, April 1, 2008; 294(4): L714 - L723. [Abstract] [Full Text] [PDF]

				O. Rentsendorj, T. Mirzapoiazova, D. Adyshev, L. E. Servinsky, T. Renne, A. D. Verin, and D. B. Pearse Role of vasodilator-stimulated phosphoprotein in cGMP-mediated protection of human pulmonary artery endothelial barrier function Am J Physiol Lung Cell Mol Physiol, April 1, 2008; 294(4): L686 - L697. [Abstract] [Full Text] [PDF]

				W.-L. Yeh, D.-Y. Lu, C.-J. Lin, H.-C. Liou, and W.-M. Fu Inhibition of Hypoxia-Induced Increase of Blood-Brain Barrier Permeability by YC-1 through the Antagonism of HIF-1{alpha} Accumulation and VEGF Expression Mol. Pharmacol., August 1, 2007; 72(2): 440 - 449. [Abstract] [Full Text] [PDF]

				G. Boerrigter, L. C. Costello-Boerrigter, A. Cataliotti, H. Lapp, J.-P. Stasch, and J. C. Burnett Jr Targeting Heme-Oxidized Soluble Guanylate Cyclase in Experimental Heart Failure Hypertension, May 1, 2007; 49(5): 1128 - 1133. [Abstract] [Full Text] [PDF]

				C. Glynos, A. Kotanidou, S. E. Orfanos, Z. Zhou, D. C. M. Simoes, C. Magkou, C. Roussos, and A. Papapetropoulos Soluble guanylyl cyclase expression is reduced in LPS-induced lung injury Am J Physiol Regulatory Integrative Comp Physiol, April 1, 2007; 292(4): R1448 - R1455. [Abstract] [Full Text] [PDF]

				A. W. Orr, R. Stockton, M. B. Simmers, J. M. Sanders, I. J. Sarembock, B. R. Blackman, and M. A. Schwartz Matrix-specific p21-activated kinase activation regulates vascular permeability in atherogenesis J. Cell Biol., February 26, 2007; 176(5): 719 - 727. [Abstract] [Full Text] [PDF]