γ-谷氨酰半胱氨酸合成酶与转录因子激活蛋白-1在吸烟气道上皮细胞表达的研究

发布时间：2018-07-07 07:23

本文选题：吸烟 + γ-谷氨酰半胱氨酸合成酶　；参考：《山西医科大学》2011年硕士论文

【摘要】：目的：(1)研究观察不同吸烟时间对大鼠支气管上皮细胞γ-谷氨酰半胱氨酸合成酶(γ-GCS)和转录因子激活蛋白-1(AP-1)表达的影响以及二者的相关性。(2)研究观察吸烟对人气道上皮细胞γ-谷氨酰半胱氨酸合成酶(γ-GCS)表达的影响,探讨γ-GCS在吸烟所致慢性气道炎症及氧化／抗氧化失衡中的作用机制。方法：(1)自制大鼠实验性被动吸烟装置,建立吸烟引起慢性气道炎症的动物模型,将40只大鼠随机分为不吸烟组、吸烟1月组、吸烟2月组、吸烟3月组,每组各10只。分别采用免疫组织化学法检测支气管上皮细胞γ-GCS和AP-1(c-fos和c-jun)蛋白的表达水平。(2)选自2007年9月-2008年10月行支气管镜检查的患者。吸烟指数300支,平均年龄50±10岁。共60例,分为4组：吸烟组(慢性支气管炎组、非慢性支气管炎组),不吸烟组(慢性支气管炎组、非慢性支气管炎组),每组各15例。通过支气管镜刷检获取支气管气道上皮细胞,采用免疫细胞化学法检测支气管上皮细胞中γ-GCS蛋白的表达水平。实验数据应用SPSSl1.0软件进行统计。结果：1.香烟对大鼠支气管上皮细胞γ-GCS表达的影响：(1)吸烟3月组大鼠出现了肺气肿的病理改变；吸烟1月、2月和3月组大鼠支气管上皮细胞γ-GCS蛋白表达的水平较不吸烟组明显增高,差异有显著性(P均0.01),(2)吸烟各组大鼠支气管上皮细胞c-fos. c-Jun蛋白的表达水平均较不吸烟组明显增高,差异有显著性(P均0.01),吸烟各组间比较差异亦有显著性(P分别0.01)；(3)吸烟1月组大鼠支气管上皮细胞c-fos与c-jun蛋白的表达水平成直线正相关(r值分别为0.679、0.735；P值分别为0.031、0.015);c-fos与γ-GCS蛋白的表达水平成直线正相关(r值分别为0.703、0.724;P值分别为0.023、0.018);c-jun与γ-GCS蛋白的表达水平成直线正相关(r值分别为0.641、0.747；P值分别为0.046、0.013)；(4)吸烟3月组大鼠支气管上皮细胞c-fos与c-jun蛋白的表达水平成直线正相关(r值分别为0.636、0.698;P值分别为0.048、0.025)；c-fos、c-jun与γ-GCS蛋白的表达水平无直线相关关系。2.吸烟对人气道上皮细胞γ-GCS表达的影响：在气管镜下取人气道上皮细胞,各组气道上皮细胞γ-GCSh的表达：(1)吸烟组(慢性支气管炎组(0.198±0.097)、非慢性支气管炎组(0.346±0.113))的气道上皮细胞γ-GCS的表达水平,低于不吸烟组(慢性支气管炎组(0.403±0.102)、非慢性支气管炎组(0.518±0.150))差异均有统计学意义(P均0.05)；(2)慢性支气管炎组(0.403±0.102)的气道上皮细胞Y-GCS的表达水平,低于非慢性支气管炎组(0.498±0.150)的表达,差异有统计学意义(P0.05)。结论：1.随着吸烟时间的延长,大鼠支气管上皮细胞中γ-GCS蛋白表达水平逐渐增高；c-fos和c-jun二者的表达也逐渐增高,并且呈正相关；2.但吸烟者人气道内上皮细胞γ-GCS的免疫活性降低,γ-GCS在不吸烟非慢性支气管炎者表达最强,而在吸烟者与慢性支气管炎者的气道内表达出现下调；γ-GCS在香烟所致慢性气道炎症、氧化应激及氧化/抗氧化失衡中可能发挥一定的作用；调控γ-GCS和维持GSH水平,对于抗氧化治疗COPD还有待与进一步研究。
[Abstract]:Objective: (1) to investigate the effects of different smoking time on the expression of gamma glutamyl cysteine synthetase (gamma -GCS) and transcription factor activator -1 (AP-1) in the bronchial epithelial cells of rats and the correlation between the two. (2) the effects of smoking on the expression of gamma glutamyl cysteine synthetase (gamma -GCS) in human airway epithelial cells were investigated and studied. The role of GCS in chronic airway inflammation and oxidative / antioxidant imbalance induced by smoking.
Methods: (1) an experimental passive smoking device of self-made rats was established to establish an animal model of chronic airway inflammation caused by smoking. 40 rats were randomly divided into non smoking group, smoking January group, smoking February group, smoking March group, and 10 rats in each group. Immunohistochemistry was used to detect the protein of gamma -GCS and AP-1 (c-fos and c-jun) in bronchial epithelial cells respectively. (2) a total of 300 smoking indexes, with an average age of 50 + 10 years, were selected from September 2007 -2008 in October. A total of 60 cases were divided into 4 groups: smoking group (chronic bronchitis group, non chronic bronchitis group), non smoking group (chronic bronchitis group, non chronic bronchitis group), 15 cases in each group. Bronchoscopic brush examination The bronchial epithelial cells were obtained and the expression of gamma -GCS protein in bronchial epithelial cells was detected by immunocytochemistry. The experimental data were calculated by SPSSl1.0 software.
Results: 1. the effect of 1. cigarettes on the expression of gamma -GCS in the bronchial epithelial cells of rats: (1) the pathological changes of emphysema appeared in the rats in the March group of smoking, and the level of the expression of gamma -GCS protein in the bronchial epithelial cells of the rats in January, February and March was significantly higher than that in the non smoking group (P 0.01), and (2) on the bronchi of the rats in each group of smoking. The expression level of c-fos. c-Jun protein in the skin cells was significantly higher than that in the non smoking group (P 0.01), and the difference in smoking groups was also significant (P, respectively 0.01). (3) the expression level of c-fos and c-jun protein in the bronchial epithelial cells of the January group of smoking rats was positively correlated (r value was 0.679,0.735; P value was 0 respectively). 31,0.015); c-fos was positively correlated with the expression level of gamma -GCS protein (r value was 0.703,0.724; P value was 0.023,0.018 respectively); c-jun and the expression level of gamma -GCS protein were straight line correlation (r value was 0.641,0.747; P values were respectively); (4) the expression water of bronchial epithelial cells in smoking rats in March Positive correlation (r value 0.636,0.698, P value respectively 0.048,0.025); c-fos, c-jun and gamma -GCS protein expression level no linear correlation between.2. smoking on human airway epithelial cell gamma -GCS expression: airway epithelial cells under the trachea, the expression of gamma -GCSh in each group of airway epithelial cells: (1) smoking group (chronic Branch) The expression level of gamma -GCS in airway epithelial cells in the tracheitis group (0.198 + 0.097) and non chronic bronchitis group (0.346 + 0.113) was lower than that in the non smoking group (0.403 + 0.102) and non chronic bronchitis group (0.518 + 0.150). (2) the airway epithelium of chronic bronchitis group (2) was (0.403 + 0.102). The expression level of Y-GCS was lower than that in non chronic bronchitis group (0.498 + 0.150), and the difference was statistically significant (P0.05).
Conclusion: 1. with the prolongation of smoking time, the expression level of gamma -GCS protein in the bronchial epithelial cells of rats increased gradually, and the expression of c-fos and c-jun two increased gradually, and was positively correlated. 2. but the immune activity of gamma -GCS in the airway epithelial cells of smokers decreased, and the expression of gamma -GCS was the strongest in non smoking non chronic bronchitis. The expression of the airway in smokers and chronic bronchitis is downregulated, and gamma -GCS may play a role in chronic airway inflammation, oxidative stress and oxidation / antioxidant imbalance caused by cigarettes, and the regulation of gamma -GCS and the maintenance of GSH levels are still to be studied for antioxidant treatment of COPD.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R363

【参考文献】