Quantitative assessment of protein-bound tyrosine nitration in airway secretions from patients with inflammatory airway disease

Hisatoshi Sugiura, Masakazu Ichinose, Masafumi Tomaki, Hiromasa Ogawa, Akira Koarai, Tomomi Kitamuro, Yuichi Komaki, Takefumi Akita, Hirohito Nishino, Shinichiro Okamoto, Takaaki Akaike, Toshio Hattori

Research output: Contribution to journalReview articlepeer-review

33 Citations (Scopus)


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 < 0.05) and patients with COPD (3.04 ± 0.36 × 10-6, p < 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 < 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. More importantly, they show that increased RNS production in the airways seems to contribute in a critical way to the pathogenesis of COPD, and that the effects of RNS in airways may differ in asthma and COPD.

Original languageEnglish
Pages (from-to)49-57
Number of pages9
JournalFree Radical Research
Issue number1
Publication statusPublished - 2004 Jan


  • 3-Nitro-tyrosine
  • Asthma
  • COPD
  • Reactive nitrogen species


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