香烟烟雾提取物经aPKC_(ι/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  

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

作　　者：江刚[1] 戴爱国 胡瑞成 

机构地区：[1]湖南省人民医院呼吸内科,湖南长沙410005 [2]湖南省老年医院湖南省老年医学研究所呼吸疾病研究室,湖南长沙410016

出　　处：《基础医学与临床》2010年第8期795-800,共6页Basic and Clinical Medicine

基　　金：湖南省科技计划(01JZY2017;05JT1023);湖南省医药卫生科研课题(B2005125;B2005292)

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-GCSmRNA,免疫荧光法检测NRF2蛋白。结果 NRF2蛋白在CSE 3 h组主要表达在胞核,其胞核蛋白高表达。p-aPKCι/ζ蛋白在CSE 3 h组显著高于对照组(P<0.05)。γ-GCS蛋白及其mRNA在CSE 3 h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 3 h组(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aPKC-NRF2调节γ-GCS表达。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）in 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 expressed after exposure to CSE for 3 hours.p-aPKCι/ζ protein significantly enhanced in CSE 3 h group than the control group（P0.05）.γ-GCS protein and mRNA were significantly increased in CSE 3 h group as compared to that ofcontrol group（P0.05）.Pretreated with aPKCι/ζ inhibitor RO318220,NRF2 protein of cytoplasm strongly expressed,p-aPKCι/ζ protein,γ-GCS protein and mRNA significantly decreased than that in CSE 3 h group（P0.05）.There is a positive correlation between NRF2 and γ-GCS and p-aPKCι/ζ,and between p-aPKCι/ζ and NRF2 and γ-GCS（P0.05）.Conclusion CSE may upregulate γ-GCS expression through aPKCι/ζ-NRF2 signaling pathway in the bronchial epithelial cells of rats.

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2 Γ-谷氨酰半胱氨酸合成酶 

分 类 号：R563.9[医药卫生—呼吸系统]