Document Detail


Depletion of nitric oxide causes cell cycle alterations, apoptosis, and oxidative stress in pulmonary cells.
MedLine Citation:
PMID:  9843847     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Nitric oxide (NO.) is important in the regulation of mitochondrial function, cell signaling, and gene expression. To elucidate how endogenous NO. regulates the function of airway epithelial cells, we used carboxy-PTIO, a hydrophilic, negatively charged NO. trap, to scavenge NO. from rat lung epithelial (RLE) and rat pleural mesothelial (RPM) cells and to determine the elicitation of cell cycle alterations, apoptosis, and oxidative stress. The reaction of NO. with PTIO causes the formation of PTI, which is measured by electron spin resonance (ESR) and is a quantitative measure of NO. formation. ESR spectroscopy revealed the production of NO. in RLE or RPM cells over a period from 1 to 24 h of exposure, indicating scavenging of NO. by PTIO. Cycle analyses in confluent RLE or RPM cells revealed two- to threefold increases in S and G2/M phases after exposure to 100-200 microM PTIO as well as increases in the fraction of cells undergoing apoptosis. Direct addition of PTI to cells failed to elicit cell cycle perturbations or apoptosis. The guanylyl cyclase inhibitor ODQ mimicked the effects of PTIO. 8-Bromo-cGMP but not 8-bromo-cAMP ameliorated the PTIO- or ODQ-mediated cell cycle perturbations and apoptosis, suggesting that cGMP-dependent pathways are involved in these cell cycle perturbations. Treatment of log-phase cells with PTIO resulted in more dramatic cell cycle perturbations compared with cells treated at confluence. Assessment of 5-bromo-2'-deoxyuridine incorporation to measure DNA synthesis demonstrated decreases in PTIO-treated compared with sham cells in addition to a cell cycle arrest in late S or G2/M phase. Last, incubation with dichlorofluorescin diacetate revealed oxidative stress in PTIO- but not in PTI-exposed RLE or RPM cells. We conclude that the depletion of endogenous NO. induces oxidative stress, cell cycle perturbations, and apoptosis. Our findings illustrate the importance of endogenous NO. in the control of cell cycle progression and survival of pulmonary and pleural cells and that a critical balance between NO. and superoxide may be necessary for these physiological events.
Authors:
Y M Janssen; R Soultanakis; K Steece; E Heerdt; R J Singh; J Joseph; B Kalyanaraman
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  The American journal of physiology     Volume:  275     ISSN:  0002-9513     ISO Abbreviation:  Am. J. Physiol.     Publication Date:  1998 Dec 
Date Detail:
Created Date:  1999-02-03     Completed Date:  1999-02-03     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  0370511     Medline TA:  Am J Physiol     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  L1100-9     Citation Subset:  IM    
Affiliation:
Department of Pathology, University of Vermont, Burlington, Vermont 05405, USA.
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MeSH Terms
Descriptor/Qualifier:
Animals
Apoptosis / drug effects,  physiology*
Benzoates / pharmacology
Cell Cycle / drug effects,  physiology
Cells, Cultured
Electron Spin Resonance Spectroscopy
Enzyme Inhibitors / pharmacology
Epithelial Cells / physiology
Fluoresceins / metabolism
Guanylate Cyclase / antagonists & inhibitors,  physiology
Imidazoles / pharmacology
Lung / cytology*,  physiology*
Nitric Oxide / antagonists & inhibitors,  metabolism*
Oxadiazoles / pharmacology
Oxidative Stress / physiology*
Pleura / cytology,  physiology
Quinoxalines / pharmacology
Rats
Grant Support
ID/Acronym/Agency:
R03-OH-03467-01/OH/NIOSH CDC HHS
Chemical
Reg. No./Substance:
0/1H-(1,2,4)oxadiazolo(4,3-a)quinoxalin-1-one; 0/Benzoates; 0/Enzyme Inhibitors; 0/Fluoresceins; 0/Imidazoles; 0/Oxadiazoles; 0/Quinoxalines; 10102-43-9/Nitric Oxide; 145757-47-7/1,3-dihydroxy-4,4,5,5-tetramethyl-2-(4-carboxyphenyl)tetrahydroimidazole; 2044-85-1/diacetyldichlorofluorescein; EC 4.6.1.2/Guanylate Cyclase

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