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.
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.
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DOI：
10.1186/s12890-016-0189-1
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发表时间：
2016-02-09
影响因子：
3.1
通讯作者：
Hirata K
中科院分区：
文献类型：
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作者：
Yamada K;Asai K;Nagayasu F;Sato K;Ijiri N;Yoshii N;Imahashi Y;Watanabe T;Tochino Y;Kanazawa H;Hirata K
已知吸烟引起的氧化应激是慢性阻塞性肺病发病机制的关键机制。核因子红细胞 2 相关因子 2 (Nrf2) 是调节抗氧化防御系统的核心转录因子。本研究的目的是比较 COPD 受试者和对照受试者中 Nrf2 的表达，并确定 Nrf2 在防止氧化应激诱导的细胞凋亡中的作用。我们在这项研究中招募了 8 名慢性阻塞性肺病受试者和 7 名对照受试者。我们通过支气管镜进行支气管刷检，并从气道中获取支气管上皮细胞。通过实时PCR和Western blotting评估支气管上皮细胞中Nrf2的表达。我们使用 caspase-3/7 激活检测试剂进行延时细胞成像测定，检查了 10% 或 15% 香烟烟雾提取物 (CSE) 诱导 A549 细胞凋亡的效果，并进行了末端脱氧核苷酸转移酶介导的 dUTP 缺口末端标记测定，以确认 A549 细胞凋亡。我们还检查了 Nrf2 敲低和 0.1、0.5 和 1.0 mM N-乙酰半胱氨酸对 CSE 诱导的细胞凋亡的影响。使用 t 检验、配对 t 检验或方差分析，然后采用 Tukey-Kramer 方法进行统计分析。 COPD受试者中Nrf2 mRNA的表达量显着低于对照受试者，且Nrf2 mRNA与包装年份呈负相关。 COPD受试者中的Nrf2蛋白显着低于对照受试者。 CSE诱导的A549细胞凋亡以时间、浓度依赖性方式增加，并且通过Nrf2敲除显着增加。 N-乙酰半胱氨酸显着改善 CSE 诱导的细胞凋亡。 COPD 患者中的 Nrf2 表达低于对照组。 Nrf2 可能对 CSE 诱导的氧化应激引起的细胞凋亡具有保护作用。这些结果表明 Nrf2 参与 COPD，并且对 COPD 患者施用抗氧化剂可能是一种基本的治疗选择。
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. 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. 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. 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.