香烟烟雾提取物经aPKCι/ζ-NRF2上调大鼠气道上皮细胞γ-谷氨酰半胱氨酸合成酶表达

基础医学与临床 ›› 2010, Vol. 30 ›› Issue (8): 795-800.

香烟烟雾提取物经aPKCι/ζ-NRF2上调大鼠气道上皮细胞γ-谷氨酰半胱氨酸合成酶表达

江刚戴爱国胡瑞成

湖南省人民医院湖南省老年医院-湖南省老年医学研究所呼吸疾病研究室

收稿日期:2009-08-24 修回日期:2009-11-24 出版日期:2010-08-05 发布日期:2010-08-05
通讯作者: 戴爱国

Cigarette smoke extracts upregulate the expression of γ－glutamylcysteine synthetase via aPKCι/ζ-NRF2 signaling pathway in the bronchial epithelial cells of rats

Gang JIANG, Ai-guo DAI, Rui-cheng HU

Hunan Provincial People's Hospital

Received:2009-08-24 Revised:2009-11-24 Online:2010-08-05 Published:2010-08-05
Contact: Ai-guo DAI,

摘要/Abstract

摘要： 目的观察香烟烟雾提取物（CSE）是否通过不典型蛋白激酶Cι/ζ (aPKCι/ζ) -红系衍生的核因子相关因子2（NRF2）信号通路调控大鼠气道上皮细胞γ-谷氨酰半胱氨酸合成酶（γ-GCS）。方法用Western blot法检测γ-GCS、NRF2和p-aPKCι/ζ蛋白，免疫细胞化学法观察γ-GCS蛋白表达，反转录聚合酶链反应（RT-PCR）法检测γ-GCS mRNA，免疫荧光法检测NRF2蛋白。结果 NRF2蛋白在CSE 3h组主要表达在胞核，其胞核蛋白高表达。p-aPKCι/ζ蛋白在CSE 3h组显著高于对照组（P＜0.05）。γ-GCS蛋白及其mRNA在CSE 3h组较对照组显著增强（P＜0.05）。预先加入aPKCι/ζ抑制剂RO813220，NRF2胞质蛋白表达增强，p-aPKCι/ζ蛋白、γ-GCS蛋白及其mRNA均明显低于CSE 3h组（均P <0.05）。相关性分析显示NRF2与γ-GCS、p-aPKCι/ζ呈正相关，p-aPKCι/ζ与NRF2、γ-GCS呈正相关（P＜0.05）。结论 CSE可能通过aPKCι/ζ-NRF2调节γ-GCS表达。

关键词: 香烟烟雾提取物, 不典型蛋白激酶Cι/ζ, 红系衍生的核因子相关因子, γ-谷氨酰半胱氨酸合成酶

Abstract: Objective To observe the effect of the signal pathway of aPKCι/ζ (antypical protein kinase Cι/ζ)-NRF2 (nuclear factor-Ｅ2 related factor) on γ-glutamylcysteine synthetase（γ-GCS）of the bronchial epithelial cells of rats after exposure to cigarette smoke extracts（CSE）. Methods γ-GCS, NRF2 and p-aPKCι/ζ proteins were semi-quantified by Western blot. The expression of γ-GCS protein was assessed by immunocytochemistry. γ-GCS mRNA expression were detected by reverse transcription-polymerase chain reaction (RT-PCR). NRF2 protein was observed by immunofluorescence. Results NRF2 protein mainly expressed in the nucleus, NRF2 protein of nucleus strongly expressed after exposure to CSE for 3 hours. p-aPKCι/ζ protein significantly enhanced in CSE 3h group than the control group（P＜0.05）. γ-GCS protein and mRNA were significantly increased in CSE 3h group as compared to the control group（P＜0.05）.Pretreated with aPKCι/ζ inhibitor RO318220, NRF2 protein of cytoplasm strongly expressed, p-aPKCι/ζ protein, γ-GCS protein and mRNA were significantly decreased than CSE3h group（P＜0.05）. The correlation analysis demonstrated that there were a positive correlation between NRF2 and γ-GCS and p- aPKCι/ζ, and between p-aPKCι/ζ and NRF2 and γ-GCS（P＜0.05）. Conclusion CSE might upregulate γ-GCS expression through aPKCι/ζ -NRF2 signaling pathway in the bronchial epithelial cells of rats.

Key words: cigarette smoke extract, antypical protein kinase Cι/ζ, Nuclear factor –Ｅ2 related factor, γ- glutamylcysteine synthetase

江刚戴爱国胡瑞成. 香烟烟雾提取物经aPKCι/ζ-NRF2上调大鼠气道上皮细胞γ-谷氨酰半胱氨酸合成酶表达[J]. 基础医学与临床, 2010, 30(8): 795-800.

Gang JIANG; Ai-guo DAI; Rui-cheng HU. Cigarette smoke extracts upregulate the expression of γ－glutamylcysteine synthetase via aPKCι/ζ-NRF2 signaling pathway in the bronchial epithelial cells of rats[J]. Basic & Clinical Medicine, 2010, 30(8): 795-800.