Nitric Oxide Synthase Isoenzyme Expression and Activity in Peripheral Lung Tissue of Patients with Chronic Obstructive Pulmonary Disease
| Autor: | James C. Hogg, Caterina Brindicci, Peter J. Barnes, Misako Ito, Sergei A. Kharitonov, Kazuhiro Ito, Mark W. Elliott |
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| Rok vydání: | 2010 |
| Předmět: |
Adult
Male Pulmonary and Respiratory Medicine Pathology medicine.medical_specialty Nitric Oxide Synthase Type II Critical Care and Intensive Care Medicine Endothelial NOS Cell Line Nitric oxide Pulmonary Disease Chronic Obstructive chemistry.chemical_compound Forced Expiratory Volume Intensive care medicine Humans Registries Aged COPD Lung biology business.industry Smoking Respiratory disease Epithelial Cells Middle Aged medicine.disease Up-Regulation respiratory tract diseases Isoenzymes Blot Nitric oxide synthase Oxidative Stress medicine.anatomical_structure Breath Tests chemistry Case-Control Studies Disease Progression biology.protein Female business |
| Zdroj: | American Journal of Respiratory and Critical Care Medicine. 181:21-30 |
| ISSN: | 1535-4970 1073-449X |
| DOI: | 10.1164/rccm.200904-0493oc |
| Popis: | Nitric oxide (NO) is increased in the lung periphery of patients with chronic obstructive pulmonary disease (COPD). However, expression of the NO synthase(s) responsible for elevated NO has not been identified in the peripheral lung tissue of patients with COPD of varying severity.Protein and mRNA expression of nitric oxide synthase type I (neuronal NOS [nNOS]), type II (inducible NOS [iNOS]), and type III (endothelial NOS [eNOS]) were quantified by Western blotting and reverse transcription-polymerase chain reaction, respectively, in specimens of surgically resected lung tissue from nonsmoker control subjects, patients with COPD of varying severity, and smokers without COPD, and in a lung epithelial cell line (A549). The effects of nitrative/oxidative stress on NOS expression and activity were also evaluated in vitro in A549 cells. nNOS nitration was quantified by immunoprecipitation and dimerization of nNOS was detected by low-temperature SDS-PAGE/Western blot in the presence of the peroxynitrite generator, 3-morpholinosydnonimine-N-ethylcarbamide (SIN1), in vitro and in vivo.Lung tissue from patients with severe and very severe COPD had graded increases in nNOS (mRNA and protein) compared with nonsmokers and normal smokers. Hydrogen peroxide (H(2)O(2)) and SIN1 as well as the cytokine mixture (IFN-gamma, IL-1beta, and tumor necrosis factor-alpha) increased mRNA expression and activity of nNOS in A549 cells in a concentration-dependent manner compared with nontreated cells. Tyrosine nitration resulted in an increase in nNOS activity in vitro, but did not affect its dimerization.Patients with COPD have a significant increase in nNOS expression and activity that reflects the severity of the disease and may be secondary to oxidative stress. |
| Databáze: | OpenAIRE |
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