Quantitative Assessment of Protein-bound Tyrosine Nitration in Airway Secretions from Patients with Inflammatory Airway Disease

Because reactive nitrogen species (RNS) have potent inflammatory activity, they may be involved in the inflammatory process in pulmonary diseases. We recently reported increased numbers of 3-nitrotyrosine immunopositive cells, which are evidences of RNS production, in the sputum of patients with chr...

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Veröffentlicht in:Free radical research 2004-01, Vol.38 (1), p.49-57
Hauptverfasser: Sugiura, Hisatoshi, Ichinose, Masakazu, Tomaki, Masafumi, Ogawa, Hiromasa, Koarai, Akira, Kitamuro, Tomomi, Komaki, Yuichi, Akita, Takefumi, Nishino, Hirohito, Okamoto, Shinichiro, Akaike, Takaaki, Hattori, Toshio
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container_title Free radical research
container_volume 38
creator Sugiura, Hisatoshi
Ichinose, Masakazu
Tomaki, Masafumi
Ogawa, Hiromasa
Koarai, Akira
Kitamuro, Tomomi
Komaki, Yuichi
Akita, Takefumi
Nishino, Hirohito
Okamoto, Shinichiro
Akaike, Takaaki
Hattori, Toshio
description Because reactive nitrogen species (RNS) have potent inflammatory activity, they may be involved in the inflammatory process in pulmonary diseases. We recently reported increased numbers of 3-nitrotyrosine immunopositive cells, which are evidences of RNS production, in the sputum of patients with chronic obstructive pulmonary disease (COPD) and patients with asthma compared with healthy subjects. In the present study, we attempted to quantify this protein nitration in the airways by means of high-performance liquid chromatography (HPLC) used together with an electrochemical detection system that we developed. Sputum samples were obtained from 15 stable COPD patients, 9 asthmatic patients and 7 healthy subjects by using hypertonic saline inhalation. The values for the molar ratio of protein-bound 3-nitrotyrosine/tyrosine in patients with asthma (4.31±1.13 × 10-6, p
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We recently reported increased numbers of 3-nitrotyrosine immunopositive cells, which are evidences of RNS production, in the sputum of patients with chronic obstructive pulmonary disease (COPD) and patients with asthma compared with healthy subjects. In the present study, we attempted to quantify this protein nitration in the airways by means of high-performance liquid chromatography (HPLC) used together with an electrochemical detection system that we developed. Sputum samples were obtained from 15 stable COPD patients, 9 asthmatic patients and 7 healthy subjects by using hypertonic saline inhalation. The values for the molar ratio of protein-bound 3-nitrotyrosine/tyrosine in patients with asthma (4.31±1.13 × 10-6, p&lt;0.05) and patients with COPD (3.04±0.36 × 10-6, p&lt;0.01) were significantly higher than those in healthy subjects (1.37±0.19 × 10-6). The levels of protein-bound 3-nitrotyrosine in the airways were not significantly different in asthmatic patients and COPD patients. A significant negative correlation was found between values for protein-bound 3-nitrotyrosine/tyrosine and % FEV1 values in patients with COPD (r=-0.53, p&lt;0.05) but not in patients with asthma. These results suggest that our HPLC-electrochemical method is useful for quantifying RNS production in human airways. 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subjects 3-Nitrotyrosine
Aged
Asthma
Asthma - metabolism
Asthma - physiopathology
Case-Control Studies
Chromatography, High Pressure Liquid - methods
COPD
Electrochemistry - methods
Female
Forced Expiratory Volume
Humans
Male
Middle Aged
Nitric Oxide - analysis
Proteins - metabolism
Pulmonary Disease, Chronic Obstructive - metabolism
Pulmonary Disease, Chronic Obstructive - physiopathology
Reactive nitrogen species
Reference Values
Sensitivity and Specificity
Sputum - metabolism
Tyrosine - analogs & derivatives
Tyrosine - analysis
Tyrosine - metabolism
title Quantitative Assessment of Protein-bound Tyrosine Nitration in Airway Secretions from Patients with Inflammatory Airway Disease
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