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Open Access 01-12-2016 | Research article

Impaired nuclear factor erythroid 2-related factor 2 expression increases apoptosis of airway epithelial cells in patients with chronic obstructive pulmonary disease due to cigarette smoking

Authors: Kazuhiro Yamada, Kazuhisa Asai, Fumihiro Nagayasu, Kanako Sato, Naoki Ijiri, Naoko Yoshii, Yumiko Imahashi, Tetsuya Watanabe, Yoshihiro Tochino, Hiroshi Kanazawa, Kazuto Hirata

Published in: BMC Pulmonary Medicine | Issue 1/2016

Abstract

Background

Cigarette smoking-induced oxidative stress is known to be a key mechanism in COPD pathogenesis. Nuclear factor erythroid 2-related factor 2 (Nrf2) is a central transcription factor that regulates the antioxidant defense system. The aim of this study was to compare Nrf2 expression in COPD subjects and control subjects, and to determine the role of Nrf2 in protecting against oxidative stress-induced apoptosis.

Methods

We enrolled 8 COPD subjects and 7 control subjects in this study. We performed bronchial brushing by bronchoscopy and obtained bronchial epithelial cells from the airways. Nrf2 expression in bronchial epithelial cells was evaluated by real-time PCR and Western blotting. We examined the effect of 10 or 15 % cigarette smoke extract (CSE) induced A549 cells apoptosis using a time-lapse cell imaging assay with caspase-3/7 activation detecting reagent and performed Terminal deoxynucleotidyltransferase-mediated dUTP nick end labelling assay for confirming A549 cells apoptosis. We also examined the effects of Nrf2 knockdown and, 0.1, 0.5, and 1.0 mM N-acetyl cysteine on CSE-induced apoptosis. Statistical analyses were performed using t-test, paired t-test or an analysis of variance followed by the Tukey-Kramer method.

Results

Nrf2 mRNA expression in COPD subjects was significantly lower than that in control subjects and Nrf2 mRNA were negatively correlated with pack year. Nrf2 protein in COPD subjects was significantly lower than that in control subjects. CSE-induced A549 cells apoptosis was increased in a time-, concentration-dependent manner, and was significantly increased by Nrf2 knockdown. N-acetyl cysteine significantly ameliorated CSE-induced apoptosis.

Conclusions

Nrf2 expression was lower in COPD patients than in control subjects. Nrf2 might have a protective role against apoptosis caused by CSE-induced oxidative stress. These results suggest an involvement of Nrf2 in COPD and administration of antioxidants to patients with COPD might be a basic therapeutic option.

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Title: Impaired nuclear factor erythroid 2-related factor 2 expression increases apoptosis of airway epithelial cells in patients with chronic obstructive pulmonary disease due to cigarette smoking
Authors: Kazuhiro Yamada
Kazuhisa Asai
Fumihiro Nagayasu
Kanako Sato
Naoki Ijiri
Naoko Yoshii
Yumiko Imahashi
Tetsuya Watanabe
Yoshihiro Tochino
Hiroshi Kanazawa
Kazuto Hirata
Publication date: 01-12-2016
Publisher: BioMed Central
Published in: BMC Pulmonary Medicine / Issue 1/2016
Electronic ISSN: 1471-2466
DOI: https://doi.org/10.1186/s12890-016-0189-1

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Results

Conclusions

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