转录因子Nrf2／Bach1及MAPK激酶调控γ-GCS在大鼠慢性阻塞性肺疾病中的作用  

Regulation of Nrf2/Bach1 on γ-glutamylcysteine synthetase in the lung tissue of chronic obstructive pulmonary disease and the effect of mitogen-activated protein kinase in rats

作　　者：王梅芳 戴爱国 胡瑞成 高军丽 朱黎明 

机构地区：[1]湖南省老年医学研究所呼吸疾病研究室,湖南省老年医院,湖南长沙410001

出　　处：《中国病理生理杂志》2009年第5期959-964,共6页Chinese Journal of Pathophysiology

基　　金：湖南省卫生厅科研基金资助项目(No.B2005125);湖南省科技计划资助项目(No.05JT1023);湖南省科技计划重点资助项目(No.2007FJ3016)

摘　　要：目的:研究转录因子红系衍生核因子相关因子-2(Nrf2)及BTB-CNC异体同源体1(Bach1)在慢性阻塞性肺疾病大鼠肺组织中的表达,探索其感受氧化应激调控谷氨酰半胱氨酸合成酶(γ-GCS)表达的可能机制。方法:复制慢性阻塞性肺疾病(COPD)模型,通过免疫组化、Westernblotting、逆转录-聚合酶链反应(RT-PCR)技术研究肺组织Nrf2、Bach1、γ-GCSmRNA及其蛋白和p-ERK、p-p38、p-JNK蛋白的表达。结果:免疫组化结果显示COPD组p-ERK、p-p38、γ-GCS、Nrf2蛋白水平较对照组升高(P<0.01),而Bach1、p-JNK蛋白水平无差异(P>0.05);Westernblotting结果显示COPD组p-ERK、p-JNK、Nrf2核蛋白质水平较对照组升高(P<0.01),Bach1、p-p38浆蛋白水平较对照组升高(P<0.01)而Bach1核蛋白水平较对照组降低(P<0.01);RT-PCR结果显示,COPD组Nrf2、Bach1mRNA水平较对照组无差异(P>0.05),而γ-GCSmRNA水平升高(P<0.01);直线相关分析结果表明,γ-GCSmRNA表达水平与Nrf2、p-ERK、p-p38总蛋白质水平,Nrf2、p-ERK、p-JNK核蛋白水平和Bach1、p-p38浆蛋白质水平均呈正相关(P<0.01),而与Bach1核蛋白质水平呈负相关(P<0.01);Nrf2核蛋白质水平与p-ERK、p-JNK核蛋白水平呈正相关(P<0.01),而Bach1核蛋白质水平与p-ERK、p-JNK核蛋白水平呈负相关(P<0.01)。结论:在慢性阻塞性肺疾病的发病过程中转录因子Nrf2/Bach1可能在核内竞争反向调节抗氧化基因γ-GCS的表达;信号转导蛋白p-ERK、p-JNK在调控Nrf2/Bach1核内平衡中起重要作用,且调控主要发生在转录后水平;γ-GCS可能还存在其它转录调节机制。AIM ： To evaluate the expression of transcriptional factors NF - E2 related factor 2 （ NrE2）, BTB and CNC homology 1 （Bachl） in rat lungs with chronic obstructive pulmonary disease （COPD）, and to explore their mechanism of regulating the expression of γ - glutamylcysteine synthetase （ γ- GCS） in oxidative state. METHODS ： The COPD model was established. Reverse transcription- polymerase chain reaction （RT- PCR）, immunohistochemistry and Western blotting techniques were used. RESULTS： Immunohistochemical results showed that p- ERK, p- p38, γ - GCS, Nrf2 protein levels were higher in COPD group than those in control group （ all P 〈 0. 01 ）. Western blotting resuits showed that p - ERK, p - JNK, Nrf2 nuclear protein expressions were higher than those in normal control group （ P 〈 0. 01 ）. In COPD group cytoplasm protein levels of p- p38 and Bachl were high （P 〈 0. 01 ） while nuclear bachl protein level was low （P 〈0. 01 ） compared to those in control group. RT- PCR results showed that Nrf2 and Bachl mRNA expressions in COPD group had no significant differences with those in control group （P 〉 0. 05 ） while γ - GCS mRNA ex-pression in COPD group was higher than that in control group （ P 〈 0.01 ）. Linear correlation analysis showed that γ- GCS mRNA was positively correlated with Nrt2, p -ERK, p -p38 total proteins, nuclear Nrf2, p -ERK, p -JNK proteins and cytoplasm Bachl, p -p38 protein while negatively correlated with bachl nuclear protein （P 〈 0. 01 ）. Bachl nuclear protein was negatively correlated with p - ERK while Bachl cytoplasm protein was positively correlated with it. Positive correlations between nuclear Nrt2 protein and nuclear p - ERK, p - JNK protein （P 〈 0. 01 ） were observed while negative correlations between nuclear Bachl protein and nuclear p - ERK, p - JNK protein （ P 〈 0.01 ） were found. CONCLUSION： In the nucleus, Nrf2 up - regulates the expression of T - GCS while Bachl down - regulates it in the physiopathol

关 键 词：肺疾病 阻塞性 红系衍生核因子相关因子-2 BTB—CNC异体同源体1 谷氨酰半胱氨酸合成 酶 有丝分裂素激活蛋白激酶类 

分 类 号：R563.3[医药卫生—呼吸系统]