摘要
为了探讨Bach1(BTBand CNC homology1)与红系衍生的核因子相关因子2(Nrf2)和γ-谷氨酰半胱氨酸合成酶(γ-GCS)的表达变化在慢性阻塞性肺疾病(COPD)中的作用和意义,用气管内注入脂多糖及熏香烟的复合刺激法建立大鼠COPD模型.观察两组大鼠的肺组织病理学改变,测两组大鼠的肺功能指标;应用免疫组化、Western印迹、原位杂交和逆转录-聚合酶链反应(RT-PCR)等方法检测Bach1与Nrf2及γ-GCS在两组大鼠的肺组织中的表达.结果显示,COPD组的肺功能指标(FEV0.3、FEV0.3/FVC%、PEF)明显恶化;光镜下肺组织病理改变符合COPD的特征性改变;γ-GCS与Nrf2的mRNA及蛋白质表达在COPD组大鼠肺组织中明显增强;而Bach1 mRNA及蛋白质在COPD组和对照组的表达无明显差别.且核/胞浆分离技术表明,Nrf2蛋白在对照组主要表达于胞浆,胞核中表达水平较弱,在COPD组胞浆、胞核均有表达,但是在胞核中的水平明显升高;Bach1蛋白在对照组主要表达于胞核中,胞浆中无明显表达,在COPD组主要表达于胞浆,胞核中无明显表达.相关性分析表明,γ-GCS mRNA与FEV0.3、FEV0.3/FVC%、PEF均呈负相关;Nrf2蛋白表达与γ-GCSmRNA呈正相关;Bach1蛋白与γ-GCS mRNA呈负相关.上述结果提示,Bach1与Nrf2和γ-GCS可能均在大鼠COPD的发病中发挥作用,且Bach1和Nrf2可能通过竞争性机制调控γ-GCS的表达,影响COPD的发生和发展.
To investigate the expression of NF-E2-related factor 2（Nrf2）, BTB and CNC homology 1 （Bachl） and γ-glutamylcysteine synthetase （7-GCS） in pulmonary tissues of rats with chronic obstructive pulmonary disease, the rat COPD model （COPD group） was established by intratracheal instillation of lipopolysaccharide （LPS） twice and exposure to cigarette smoke. The lung function measurements FEV0.3, FEV0.3/FVC% and PEF were carried out. The pathological changes were also observed. In situ hybridization（ISH）, reverse transcription-polymerase chain reaction （RT-PCR）, immunohistochemistry and Western blot were used to detect the expression of Bach, Nrf2 and γ-GCS. The results showed that the lung function of COPD group was significantly aggravated. The COPD group rats shared specific pathological features of COPD. The levels of γ- GCS and Nrf2 mRNA and protein in COPD group were significantly higher than those in control group. The levels of Bachl mRNA and protein in COPD group and control group were stable. Nuclear/cytosol fractionation showed that Nrf2 protein was mainly expressed in cytoplasm in control group. But Nrf2 protein was mainly expressed in nucleus. Bachl protein of cytoplasm was mainly detected in COPD group. Linear correlation analysis showed that γ-GCS mRNA was negatively correlated with FEV0.3, FEV0.3/FVC% and PEF. Nrf2 protein was positively correlated with γ-GCS mRNA. Bachl protein was negatively correlated with γ-GCS mRNA. Taken together, these findings indicate that Bachl, Nrf2 and γ-GCS may play important roles in the development of COPD.
出处
《中国生物化学与分子生物学报》
CAS
CSCD
北大核心
2008年第7期649-655,共7页
Chinese Journal of Biochemistry and Molecular Biology
基金
湖南省科技计划项目(No05JT1023)
湖南省医药卫生科研课题(NoB2005125
2005292)资助~~
