大鼠慢性阻塞性肺疾病中P-ERk参与Bach1调控γ-GCS的表达  

P-ERK was Involved in the Expression of γ-Glutamylcysteine Synthetase Regulated by Bach1 in Rats with Chronic Obstructive Pulmonary Disease

作　　者：王梅芳[1,2] 戴爱国 胡瑞成 朱黎明 

机构地区：[1]南华大学研究生院,湖南衡阳421001 [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410001

出　　处：《郧阳医学院学报》2008年第5期385-389,F0002,共6页Journal of Yunyang Medical College

基　　金：湖南省卫生厅科研项目(B2005125);湖南省科技计划项目(05JT1023);湖南省卫生厅科研计划项目(B2007169)

摘　　要：目的:探讨慢性阻塞性肺疾病(COPD)中BTB-CNC异体同源体1(BTBandCNChomology1,Bach1)对谷氨酰半胱氨酸连接酶(γ-GCS)表达的调控及磷酸化的细胞外信号调节激酶(P-ERK)可能的参与作用。方法:复制COPD模型,通过免疫组化、westernblot、RT-PCR检测肺组织Bach1、γ-GCSmRNA及其蛋白和P-ERK蛋白的表达。结果:免疫组化结果显示COPD组P-ERK、γ-GCS蛋白水平较对照组升高(P<0.01),而Bach1蛋白水平无差异(P>0.05);western印迹结果显示COPD组Bach1核蛋白水平较对照组降低(P<0.01),而浆蛋白水平升高(P<0.01);RT-PCR结果显示COPD组Bach1mRNA水平较对照组无差异(P>0.05),而γ-GCSmRNA水平升高(P<0.01);相关分析发现γ-GCSmRNA及蛋白的表达与P-ERK蛋白、Bach1浆蛋白呈正相关(P<0.01),而与Bach1核蛋白呈负相关(P<0.01);Bach1核蛋白与P-ERK蛋白呈负相关(P<0.01),而Bach1浆蛋白与P-ERK蛋白呈正相关(P<0.01)。结论:Bach1与P-ERK均参与了大鼠COPD的发病过程:Bach1在核内下调抗氧化基因γ-GCS的表达,氧化应激时P-ERK介导其出核,上调抗氧化基因γ-GCS的表达;P-ERK主要在转录后水平调节Bach1胞浆胞核穿梭运动。Objective To explore the regulation of BTB and CNC homologyl （ Bachl ） on γ-glutamylcysteine synthetase （ γ- GCS） expression in chronic obstructive pulmonary disease （COPD） and the role of phosphorylation of extraceUular signalregulated kinase （P-ERK） for γ-GCS expression regulated by Bachl. Methods The COPD model was established. Reverse transcriptiun-polymerase chain reaction （RT-PCR） , immunohistochemistry and Western blot were used to determine the contents of Bachl, γ-GCS mRNA and P-ERK protein in the lung tissues. Results Immunohistochemistry showed that P- ERK,γ-GCS protein levels were higher in COPD group than that in control group （ all P 〈0.01 ） ,but the level of Bachl had no significant difference（ P 〉0.05 ）. Western blot showed that in COPD group Bachl cytoplasm protein level was increased （ P 〈 0.01 ）, while Bachl nuclear protein level was decreased（ P 〈 0. 01 ） compared with control group. The expression of Bachl mRNA in COPD group had no significant difference with that in control group （ P 〉 0.05 ） , while γ-GCS mRNA expression in COPD group was higher than that in control group（ P 〈 0. 01 ）. Linear correlation analysis showed that γ- GCS mRNA and protein were positively correlated with P-ERK and Bachl cytoplasm protein, while negatively correlated with Bach1 nuclear protein（ all P 〈 0.01 ） ;Bachl nuclear protein was negatively correlated with P-ERK,while Bachl cytoplasm protein was positively correlated with P-ERK. Conclusion Both Bachl and P-ERK participate the pathogenesis of COPD in rats, Bachl could down-regulate the expression of γ-GCS in the nucleus, P-ERK can mediate the export of Bachl in the oxidative stress state, up-regulate γ-GCS expression, Bachl is mainly regulated by P-ERK at the post-transcriptional level.

关 键 词：肺疾病 阻塞性 BTB-CNC异体同源体1 谷氨酰半胱氨酸连接酶 磷酸化的细胞外信号调节激酶 

分 类 号：R563.9[医药卫生—呼吸系统]