【摘要】：研究背景慢性阻塞性肺疾�。╟hronicobstructivepulmonarydisease，COPD）是呼吸系統(tǒng)常見和多發(fā)的疾病，患病率和死亡率均較高，其中吸煙是重要的危險因素。目前認為COPD的發(fā)病機制主要包括以下三個方面:1、氣道和肺部炎癥;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發(fā)病中的作用日益受到重視。4-羥基-壬烯醛（4-HNE）是脂質(zhì)過氧化終末產(chǎn)物之一，研究表明4-HNE有調(diào)節(jié)抗氧化物質(zhì)谷胱甘肽，參與氧化抗氧化失衡，炎癥反應(yīng)，細胞凋亡，信號傳導(dǎo)的作用，還能導(dǎo)致血管水腫，內(nèi)皮細胞滲透性改變，影響基因的表達。谷胱甘肽是肺部抗氧化的重要屏障，可以保護細胞免受4-HNE的損傷。γ-谷氨酰半胱氨酸合成酶（γ-GCS）是谷胱甘肽合成過程中的限速酶,它的數(shù)量、活性等直接影響谷胱甘肽的水平。本課題旨在探討4-HNE及γ-GCS在COPD氧化抗氧化失衡發(fā)病機制中的作用。 目的通過觀察不同吸煙量、吸煙持續(xù)時間對大鼠支氣管肺泡灌洗液（BALF）及外周血4-HNE和γ-GCS表達的影響及其相關(guān)性，探討4-HNE及γ-GCS在COPD氧化抗氧化失衡發(fā)病機制中的作用。方法實驗動物雄性Wistar大鼠30只，隨機分為不吸煙組、吸煙6周組、吸煙12周組。吸煙6周及12周組：每次吸煙15支，大約2小時，，每天吸煙2次（上、下午各一次），每周吸煙6天，分別吸煙6、12周，不吸煙組大鼠正常飼養(yǎng)12周，與吸煙組飼養(yǎng)條件及環(huán)境相同。采用雙抗體夾心酶聯(lián)免疫吸附（ABC-ELISA）法分別檢測各組大鼠BALF及外周血4-HNE和γ-GCS的含量，并分析兩者的相關(guān)性。 結(jié)果1、4-HNE的表達含量較之不吸煙組(15.917±1.584；12.305±1.822)，吸煙6周組(21.376±2.80；18.333±2.585)和吸煙12周組(28.634±2.999；22.754±3.882)大鼠BALF和外周血中4-HNE含量明顯增加，差異均具有統(tǒng)計學意義(P均＜0.01)；吸煙12周組較之吸煙6周組有所增加，差異也存在統(tǒng)計學意義(P＜0.01)。2、γ-GCS的表達含量較之不吸煙組(13.525±2.311；11.847±1.171)，吸煙6周組(15.802±1.647；16.221±1.577)大鼠BALF和外周血中γ-GCS的表達明顯增加，差異均存在統(tǒng)計學意義(P0.05；P＜0.01)；較之不吸煙組比較，吸煙12周組(11.527±1.606；9.015±1.569)的表達明顯減少，差異也存在統(tǒng)計學意義(P＜0.05；P＜0.01)。3、吸煙6周組BALF及外周血4-HNE及γ-GCS的表達呈正相關(guān)（r=0.764，r=0.674），吸煙12周組BALF中4-HNE及γ-GCS表達呈負相關(guān)（r=㧟0.777），差異均存在統(tǒng)計學意義（P均＜0.05），而外周血中二者表達無顯著相關(guān)性。 結(jié)論香煙煙霧暴露可使大鼠體液4-HNE及γ-GCS的表達水平增高，隨著吸煙時間延長，γ-GCS逐漸降低，提示二者在COPD氧化抗氧化失衡中發(fā)揮一定的作用。
[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. 緯 -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 緯 -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 緯 -GCS in bronchoalveolar lavage fluid (BALF) and peripheral blood of rats and to explore the role of 4-HNE and 緯 -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 緯 -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 鹵1.584 鹵12.305 鹵1.822), smoking group of 6 weeks (21.376 鹵2.80,18.333 鹵2.585) and smoking group of 12 weeks (28.634 鹵2.9990.79 鹵22.754 鹵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 鹵2.80,18.333 鹵2.585) and 12-week smoking group (28.634 鹵2.999 鹵22.754 鹵3.882). The expression of 緯 -GCS in the 12-week smoking group was significantly higher than that in the non-smoking group (13.525 鹵2.311 鹵11.847 鹵1.171), and the expression of BALF and 緯 -GCS in peripheral blood was significantly increased in the 6-week smoking group (15.802 鹵1.647 鹵16.221 鹵1.577). The difference was statistically significant (P < 0. 05), and the expression of 12 weeks smoking group (11.527 鹵1. 606 鹵9. 015 鹵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 緯 -GCS in peripheral blood of 6 weeks smoking group (r = 0. 764), and a negative correlation between the expression of 4-HNE and 緯 -GCS in BALF of 12 weeks smoking group (r = 0. 777), but there was no significant correlation between the expression of 4-HNE and 緯 -GCS in peripheral blood (P < 0. 05). Conclusion cigarette smoke exposure can increase the expression of 4-HNE and 緯 -GCS in the body fluid of rats. 緯 -GCS decreases gradually with the prolongation of smoking time, suggesting that they play a role in the imbalance of oxidation and antioxidation of COPD.
【學位授予單位】：山西醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2012
【分類號】：R563.9

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