香烟烟雾暴露对大鼠4-HNE和γ-GCS表达的影响
发布时间：2018-09-04 17:56
【摘要】：研究背景慢性阻塞性肺疾病（chronicobstructivepulmonarydisease，COPD）是呼吸系统常见和多发的疾病，患病率和死亡率均较高，其中吸烟是重要的危险因素。目前认为COPD的发病机制主要包括以下三个方面:1、气道和肺部炎症;2、蛋白酶/抗蛋白酶失衡;3、氧化/抗氧化失衡。其中氧化/抗氧化失衡在COPD发病中的作用日益受到重视。4-羟基-壬烯醛（4-HNE）是脂质过氧化终末产物之一，研究表明4-HNE有调节抗氧化物质谷胱甘肽，参与氧化抗氧化失衡，炎症反应，细胞凋亡，信号传导的作用，还能导致血管水肿，内皮细胞渗透性改变，影响基因的表达。谷胱甘肽是肺部抗氧化的重要屏障，可以保护细胞免受4-HNE的损伤。γ-谷氨酰半胱氨酸合成酶（γ-GCS）是谷胱甘肽合成过程中的限速酶,它的数量、活性等直接影响谷胱甘肽的水平。本课题旨在探讨4-HNE及γ-GCS在COPD氧化抗氧化失衡发病机制中的作用。 目的通过观察不同吸烟量、吸烟持续时间对大鼠支气管肺泡灌洗液（BALF）及外周血4-HNE和γ-GCS表达的影响及其相关性，探讨4-HNE及γ-GCS在COPD氧化抗氧化失衡发病机制中的作用。方法实验动物雄性Wistar大鼠30只，随机分为不吸烟组、吸烟6周组、吸烟12周组。吸烟6周及12周组：每次吸烟15支，大约2小时，，每天吸烟2次（上、下午各一次），每周吸烟6天，分别吸烟6、12周，不吸烟组大鼠正常饲养12周，与吸烟组饲养条件及环境相同。采用双抗体夹心酶联免疫吸附（ABC-ELISA）法分别检测各组大鼠BALF及外周血4-HNE和γ-GCS的含量，并分析两者的相关性。 结果1、4-HNE的表达含量较之不吸烟组(15.917±1.584；12.305±1.822)，吸烟6周组(21.376±2.80；18.333±2.585)和吸烟12周组(28.634±2.999；22.754±3.882)大鼠BALF和外周血中4-HNE含量明显增加，差异均具有统计学意义(P均＜0.01)；吸烟12周组较之吸烟6周组有所增加，差异也存在统计学意义(P＜0.01)。2、γ-GCS的表达含量较之不吸烟组(13.525±2.311；11.847±1.171)，吸烟6周组(15.802±1.647；16.221±1.577)大鼠BALF和外周血中γ-GCS的表达明显增加，差异均存在统计学意义(P0.05；P＜0.01)；较之不吸烟组比较，吸烟12周组(11.527±1.606；9.015±1.569)的表达明显减少，差异也存在统计学意义(P＜0.05；P＜0.01)。3、吸烟6周组BALF及外周血4-HNE及γ-GCS的表达呈正相关（r=0.764，r=0.674），吸烟12周组BALF中4-HNE及γ-GCS表达呈负相关（r=㧟0.777），差异均存在统计学意义（P均＜0.05），而外周血中二者表达无显著相关性。 结论香烟烟雾暴露可使大鼠体液4-HNE及γ-GCS的表达水平增高，随着吸烟时间延长，γ-GCS逐渐降低，提示二者在COPD氧化抗氧化失衡中发挥一定的作用。
[Abstract]:Background chronic obstructive pulmonary disease (chronicobstructivepulmonarydisease,COPD) is a common and frequent respiratory disease with high morbidity and mortality. Smoking is an important risk factor. It is believed that the pathogenesis of COPD mainly includes the following three aspects: 1: 1, airway and lung inflammation 2, protease / antiproteinase imbalance 3, oxidation / antioxidation imbalance 3. The role of oxidation / antioxidation imbalance in the pathogenesis of COPD has been paid more and more attention to. 4- hydroxy-nonene aldehyde (4-HNE) is one of the end products of lipid peroxidation. It has been shown that 4-HNE regulates the antioxidant substance glutathione and participates in oxidative antioxidant imbalance. Inflammation, apoptosis and signal transduction can also lead to vascular edema, endothelial cell permeability changes, affecting gene expression. Glutathione is an important barrier of lung antioxidation, which can protect cells from 4-HNE damage. 纬 -GCS is a rate-limiting enzyme in the process of glutathione synthesis. Its quantity and activity directly affect the level of glutathione. The purpose of this study was to investigate the role of 4-HNE and 纬 -GCS in the pathogenesis of oxidative and antioxidant imbalance in COPD. Objective to investigate the effect of different amount of smoking and duration of smoking on the expression of 4-HNE and 纬 -GCS in bronchoalveolar lavage fluid (BALF) and peripheral blood of rats and to explore the role of 4-HNE and 纬 -GCS in the pathogenesis of oxidative and antioxidant imbalance of COPD. Methods Thirty male Wistar rats were randomly divided into non smoking group, 6 week smoking group and 12 week smoking group. Smoking for 6 weeks and 12 weeks: 15 cigarettes, about 2 hours, 2 times a day (upper, 1 in the afternoon), 6 days a week, 6 weeks, 12 weeks respectively, the rats in the non-smoking group were fed for 12 weeks. The feeding conditions and environment were the same as those in smoking group. Double antibody sandwich enzyme-linked immunosorbent assay (ABC-ELISA) was used to detect the contents of BALF, 4-HNE and 纬 -GCS in peripheral blood of rats in each group, and the correlation between them was analyzed. Results 1the expression of 4-HNE was significantly higher than that of non-smoking group (15.917 卤1.584 卤12.305 卤1.822), smoking group of 6 weeks (21.376 卤2.80,18.333 卤2.585) and smoking group of 12 weeks (28.634 卤2.9990.79 卤22.754 卤3.882). The difference was statistically significant (all P < 0.01), and the level of 4-HNE in peripheral blood was significantly higher than that in non-smoking group (21.376 卤2.80,18.333 卤2.585) and 12-week smoking group (28.634 卤2.999 卤22.754 卤3.882). The expression of 纬 -GCS in the 12-week smoking group was significantly higher than that in the non-smoking group (13.525 卤2.311 卤11.847 卤1.171), and the expression of BALF and 纬 -GCS in peripheral blood was significantly increased in the 6-week smoking group (15.802 卤1.647 卤16.221 卤1.577). The difference was statistically significant (P < 0. 05), and the expression of 12 weeks smoking group (11.527 卤1. 606 卤9. 015 卤1.569) was significantly lower than that of non smoking group (P < 0. 05). There was a positive correlation between the expression of BALF and 4-HNE and 纬 -GCS in peripheral blood of 6 weeks smoking group (r = 0. 764), and a negative correlation between the expression of 4-HNE and 纬 -GCS in BALF of 12 weeks smoking group (r = 0. 777), but there was no significant correlation between the expression of 4-HNE and 纬 -GCS in peripheral blood (P < 0. 05). Conclusion cigarette smoke exposure can increase the expression of 4-HNE and 纬 -GCS in the body fluid of rats. 纬 -GCS decreases gradually with the prolongation of smoking time, suggesting that they play a role in the imbalance of oxidation and antioxidation 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】：2012
【分类号】：R563.9
本文编号：2222913
[Abstract]:Background chronic obstructive pulmonary disease (chronicobstructivepulmonarydisease,COPD) is a common and frequent respiratory disease with high morbidity and mortality. Smoking is an important risk factor. It is believed that the pathogenesis of COPD mainly includes the following three aspects: 1: 1, airway and lung inflammation 2, protease / antiproteinase imbalance 3, oxidation / antioxidation imbalance 3. The role of oxidation / antioxidation imbalance in the pathogenesis of COPD has been paid more and more attention to. 4- hydroxy-nonene aldehyde (4-HNE) is one of the end products of lipid peroxidation. It has been shown that 4-HNE regulates the antioxidant substance glutathione and participates in oxidative antioxidant imbalance. Inflammation, apoptosis and signal transduction can also lead to vascular edema, endothelial cell permeability changes, affecting gene expression. Glutathione is an important barrier of lung antioxidation, which can protect cells from 4-HNE damage. 纬 -GCS is a rate-limiting enzyme in the process of glutathione synthesis. Its quantity and activity directly affect the level of glutathione. The purpose of this study was to investigate the role of 4-HNE and 纬 -GCS in the pathogenesis of oxidative and antioxidant imbalance in COPD. Objective to investigate the effect of different amount of smoking and duration of smoking on the expression of 4-HNE and 纬 -GCS in bronchoalveolar lavage fluid (BALF) and peripheral blood of rats and to explore the role of 4-HNE and 纬 -GCS in the pathogenesis of oxidative and antioxidant imbalance of COPD. Methods Thirty male Wistar rats were randomly divided into non smoking group, 6 week smoking group and 12 week smoking group. Smoking for 6 weeks and 12 weeks: 15 cigarettes, about 2 hours, 2 times a day (upper, 1 in the afternoon), 6 days a week, 6 weeks, 12 weeks respectively, the rats in the non-smoking group were fed for 12 weeks. The feeding conditions and environment were the same as those in smoking group. Double antibody sandwich enzyme-linked immunosorbent assay (ABC-ELISA) was used to detect the contents of BALF, 4-HNE and 纬 -GCS in peripheral blood of rats in each group, and the correlation between them was analyzed. Results 1the expression of 4-HNE was significantly higher than that of non-smoking group (15.917 卤1.584 卤12.305 卤1.822), smoking group of 6 weeks (21.376 卤2.80,18.333 卤2.585) and smoking group of 12 weeks (28.634 卤2.9990.79 卤22.754 卤3.882). The difference was statistically significant (all P < 0.01), and the level of 4-HNE in peripheral blood was significantly higher than that in non-smoking group (21.376 卤2.80,18.333 卤2.585) and 12-week smoking group (28.634 卤2.999 卤22.754 卤3.882). The expression of 纬 -GCS in the 12-week smoking group was significantly higher than that in the non-smoking group (13.525 卤2.311 卤11.847 卤1.171), and the expression of BALF and 纬 -GCS in peripheral blood was significantly increased in the 6-week smoking group (15.802 卤1.647 卤16.221 卤1.577). The difference was statistically significant (P < 0. 05), and the expression of 12 weeks smoking group (11.527 卤1. 606 卤9. 015 卤1.569) was significantly lower than that of non smoking group (P < 0. 05). There was a positive correlation between the expression of BALF and 4-HNE and 纬 -GCS in peripheral blood of 6 weeks smoking group (r = 0. 764), and a negative correlation between the expression of 4-HNE and 纬 -GCS in BALF of 12 weeks smoking group (r = 0. 777), but there was no significant correlation between the expression of 4-HNE and 纬 -GCS in peripheral blood (P < 0. 05). Conclusion cigarette smoke exposure can increase the expression of 4-HNE and 纬 -GCS in the body fluid of rats. 纬 -GCS decreases gradually with the prolongation of smoking time, suggesting that they play a role in the imbalance of oxidation and antioxidation 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】：2012
【分类号】：R563.9
【参考文献】
相关期刊论文 前10条
1 宋宁;COPD与细胞凋亡[J];国外医学.呼吸系统分册;2001年04期
2 何志义;氧化/抗氧化失衡与慢性阻塞性肺疾病[J];国外医学(呼吸系统分册);2003年01期
3 朱渊红;氧化/抗氧化失衡与慢性阻塞性肺疾病[J];国外医学(呼吸系统分册);2003年01期
4 陈娟;冉丕鑫;;4-羟基壬烯醛在疾病发生机制方面的研究进展[J];国际呼吸杂志;2006年11期
5 赵立敏;刘峰;史苏芹;杨秀芬;;氧化应激及抗氧化治疗对慢性阻塞性肺疾病的影响[J];临床荟萃;2009年04期
6 许建英;孙丽娜;庞敏;;γ-谷氨酰半胱氨酸合成酶、转化生长因子β1在吸烟大鼠肺组织中的表达[J];山西医科大学学报;2009年02期
7 于亮;冉丕鑫;;4-羟基壬烯醛诱导支气管上皮细胞凋亡[J];现代临床医学生物工程学杂志;2006年05期
8 韩玲;许建英;姚宏;;γ-谷氨酰半胱氨酸合成酶在吸烟气道上皮细胞表达的研究[J];中国药物与临床;2011年03期
9 邹朝霞,何志义,冉丕鑫;香烟烟雾提取物影响肺泡上皮细胞谷胱甘肽和γ谷氨酰半胱氨酸合成酶的表达[J];中华结核和呼吸杂志;2005年06期
10 于亮;冉丕鑫;;香烟对气道上皮细胞表达4-羟基壬烯醛的影响以及银杏内酯B的干预作用[J];中华结核和呼吸杂志;2006年04期
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