help button home button
AJRCMB
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Zulueta, J. J.
Right arrow Articles by Hassoun, P. M.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Zulueta, J. J.
Right arrow Articles by Hassoun, P. M.

Am. J. Respir. Cell Mol. Biol., Volume 26, Number 1, January, 2002 22-30

Modulation of Inducible Nitric Oxide Synthase by Hypoxia in Pulmonary Artery Endothelial Cells

Javier J. Zulueta, Raneeta Sawhney, Usamah Kayyali, Michael Fogel, Cameron Donaldson, Hailu Huang, Joseph J. Lanzillo, and Paul M. Hassoun

Pulmonary and Critical Care Division, Department of Medicine/Tupper Research Institute, New England Medical Center, Boston, Massachusetts; and Tufts University School of Medicine, Boston, Massachusetts

The effects of hypoxia on the regulation of inducible nitric oxide synthase (NOS) 2 expression were examined in cultured rat pulmonary microvascular endothelial cells (EC). EC did not express NOS 2 mRNA or protein when exposed to normoxia or hypoxia unless they were pretreated with interleukin (IL)-1beta and/or tumor necrosis factor (TNF)-alpha for 24 h. Induction of NOS 2 by IL-1beta +TNF-alpha was significantly attenuated by concomitant exposure of EC to hypoxia or treatment of EC with antioxidants such as tiron, diphenyliodonium, and catalase, suggesting that NOS 2 expression is dependent on the production of reactive oxygen species. Degradation of Ikappa B and activation of NF-kappa B, which were both induced by treatment of EC with cytokines, were not altered when the cells were exposed to hypoxia, suggesting that the modulation of NOS 2 expression by hypoxia is unrelated to NF-kappa B activation. Following stimulation with IL-1beta +TNF-alpha for 24 h, incubation of EC in normoxia resulted in a progressive decline in NOS 2 expression and a calculated half-life of approximately 6 h for NOS 2 mRNA. Hypoxia significantly prolonged the half-life of NOS 2 mRNA (17 h, P < 0.05 versus normoxic EC). The half-life of NOS 2 mRNA was also prolonged by actinomycin D treatment (19.5 and 29.5 h for normoxic and hypoxic EC, respectively), suggesting that transcription of an RNA destabilizing factor or RNAse contributes to NOS 2 mRNA degradation. In EC transiently transfected with the rat NOS 2 promoter, hypoxia and the combination of IL-1beta +TNF-alpha independently increased promoter activity 2.2- and 3-fold, respectively. As opposed to the attenuating effect that hypoxia had on IL-1beta +TNF-alpha - dependent induction of NOS 2 gene expression, the concomitant treatment with IL-1beta +TNF-alpha and hypoxia synergistically increased NOS 2 promoter activity 17.6-fold. Taken together, these results suggest that hypoxia alone does not induce NOS 2 expression in cultured pulmonary microvascular EC, but may modulate cytokine induction of this enzyme at pretranscriptional, transcriptional, and posttranscriptional levels.

Abbreviations: diaminonaphtalene, DAN; endothelial cell, EC; ethylenediaminetetraacetic acid, EDTA; hypoxia-inducible factor, HIF; interleukin, IL; inducible nitric oxide synthase, iNOS; internal standard, IS; Nomega -nitro-L -arginine methyl esther, L-NAME; Moloney murine leukemia virus reverse transcriptase, MMLV-RT; nitric oxide, NO; nitric oxide synthase, NOS; 1-(H)-naphtotriazole, NT; polymerase chain reaction, PCR; reactive oxygen species, ROS; tumor necrosis factor, TNF; tumor necrosis factor-alpha , TNF-alpha .




This article has been cited by other articles:


Home page
 Am. J. Physiol. Cell Physiol.Home page
M. A. Robinson, J. E. Baumgardner, V. P. Good, and C. M. Otto
Physiological and hypoxic O2 tensions rapidly regulate NO production by stimulated macrophages
Am J Physiol Cell Physiol, April 1, 2008; 294(4): C1079 - C1087.
[Abstract] [Full Text] [PDF]

Home page
 Am. J. Respir. Crit. Care Med.Home page
X. Peng, R.-E. E. Abdulnour, S. Sammani, S.-F. Ma, E. J. Han, E. J. Hasan, R. Tuder, J. G. N. Garcia, and P. M. Hassoun
Inducible Nitric Oxide Synthase Contributes to Ventilator-induced Lung Injury
Am. J. Respir. Crit. Care Med., August 15, 2005; 172(4): 470 - 479.
[Abstract] [Full Text] [PDF]

Home page
 Am. J. Physiol. Lung Cell. Mol. Physiol.Home page
J. Agorreta, J. J. Zulueta, L. M. Montuenga, and M. Garayoa
Adrenomedullin expression in a rat model of acute lung injury induced by hypoxia and LPS
Am J Physiol Lung Cell Mol Physiol, March 1, 2005; 288(3): L536 - L545.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
R. M. Day, G. Thiel, J. Lum, R. D. Chevere, Y. Yang, J. Stevens, L. Sibert, and B. L. Fanburg
Hepatocyte Growth Factor Regulates Angiotensin Converting Enzyme Expression
J. Biol. Chem., March 5, 2004; 279(10): 8792 - 8801.
[Abstract] [Full Text] [PDF]

Home page
 Am. J. Respir. Crit. Care Med.Home page
J. Agorreta, M. Garayoa, L. M. Montuenga, and J. J. Zulueta
Effects of Acute Hypoxia and Lipopolysaccharide on Nitric Oxide Synthase-2 Expression in Acute Lung Injury
Am. J. Respir. Crit. Care Med., August 1, 2003; 168(3): 287 - 296.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
U. S. Kayyali, C. M. Pennella, C. Trujillo, O. Villa, M. Gaestel, and P. M. Hassoun
Cytoskeletal Changes in Hypoxic Pulmonary Endothelial Cells Are Dependent on MAPK-activated Protein Kinase MK2
J. Biol. Chem., November 1, 2002; 277(45): 42596 - 42602.
[Abstract] [Full Text] [PDF]

Home page
 Am. J. Respir. Cell Mol. Bio.Home page
B. R. Pitt and C. M. St. Croix
Complex Regulation of iNOS in Lung
Am. J. Respir. Cell Mol. Biol., January 1, 2002; 26(1): 6 - 9.
[Full Text] [PDF]


HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
Proc. Am. Thorac. Soc. Am. J. Respir. Crit. Care Med.
Copyright © 2002 American Thoracic Society.