氨溴索对哮喘大鼠肺组织激活蛋白-1和γ-谷胺酰半胱氨酸合成酶表达的影响

发布时间：2018-02-26 15:23

本文关键词： 氨溴索支气管哮喘激活蛋白-1 γ-谷胺酰半胱氨酸合成酶　出处：《新乡医学院》2012年硕士论文　论文类型：学位论文

【摘要】：背景经典理论认为,哮喘是以嗜酸粒细胞(Eosinophil,EOS)浸润为主的气道炎症,但近年来很多研究发现,氧化／抗氧化失衡在哮喘的发生发展中也起着重要作用。氨溴索(ambroxol,AMB)是一种临床常用祛痰药,近年来研究发现,其具有一定抗氧化作用,在哮喘治疗中具有一定的应用价值。目的通过观察氨溴索对哮喘大鼠肺组织病理的改变以及对哮喘大鼠肺组织及支气管肺泡灌洗液中激活蛋白-1(activator protein-1,AP-1)和γ-谷胺酰半胱氨酸合成酶(y-glutamylcysteine synthetase, y-GCS)蛋白的影响,初步探讨氨溴索对哮喘大鼠气道氧化/抗氧化系统的作用及其可能的作用机制。方法将30只雄性Sprauge-Dawley(SD)大鼠随机分为3组：空白对照组、哮喘模型组和氨溴索处理组。哮喘模型组和氨溴索处理组分别于第1d和第8d向大鼠腹腔内注射抗原液1ml,(含有卵清蛋白(ovalbumin,OVA)100mg,氢氧化铝100mg)致敏。第15天开始雾化吸入1%OVA (w/v)激发气道反应,每次20分钟,每日1次,共7天。空白对照组则于第1、8天腹腔内注射生理盐水1ml,第15天开始雾化吸入生理盐水,每次20分钟,每日1次,共7天。氨溴索处理组致敏同时给予腹腔注射氨溴索50mg.kg-1.d-1,空白对照组和哮喘模型组则同时给予等量生理盐水腹腔注射,共21天。各组动物于最后一次雾化吸入后24小时行支气管肺泡灌洗,收集支气管肺泡灌洗液(bronchoalveolar lavage fluid, BALF)。采用HE染色观察肺组织病理的改变,免疫细胞化学染色法检测支气管肺泡灌洗液中AP-1(c-Jun和c-Fos亚单位)及y-GCS的表达水平以及免疫组织化学法观察AP-1(c-Jun和c-Fos亚单位)及y-GCS在肺组织中的表达。结果(1)激发过程中哮喘模型组大鼠出现呼吸窘迫症状(呼吸急促、青紫等),氨溴索处理组大鼠激发的症状与哮喘模型组相比有明显减轻,空白对照组大鼠未见呼吸窘迫等症状。 (2)各组大鼠肺组织病理变化：空白对照组显示大鼠肺组织可见肺泡大小均一且肺泡壁薄,支气管上皮完整,周围几乎无炎性细胞浸润及其分泌物。哮喘模型组显示支气管粘膜上皮水肿伴有局部的脱落,基底膜的增厚,粘液腺增多,管壁的各层均可见嗜酸性粒细胞、巨噬细胞和单核细胞的浸润。氨溴索处理组显示病变范围、程度均较模型组明显减轻； (3)肺组织和支气管肺泡灌洗液AP-1(c-Jun、c-Fos亚单位)蛋白表达：哮喘模型组肺组织及支气管肺泡灌洗液中c-Jun蛋白和c-Fos蛋白的表达水平与空白对照组比较明显升高(P0.01)；氨溴索处理组c-Jun蛋白和c-Fos蛋白的表达水平与哮喘模型组比较明显降低(P0.01),但仍高于空白对照组(P0.01)； (4)肺组织和支气管肺泡灌洗液γ-GCS蛋白表达：哮喘模型组肺组织及支气管肺泡灌洗液γ-GCS蛋白的表达水平与空白对照组比较明显升高(P0.01)；氨溴索处理组肺组织及支气管肺泡灌洗液中γ-GCS蛋白的表达水平与哮喘模型组比较明显降低(P0.01),但仍高于空白对照组(P0.01)。结论(1)氨溴索发挥抗氧化作用,其机制可能是通过降低AP-1(c-Jun、c-Fos亚单位)和γ-GCS蛋白的表达水平有关。 (2)氨溴索通过其抗氧化作用,减轻支气管肺组织的损伤,对哮喘大鼠模型具有一定的气道保护作用。
[Abstract]:That is the background of classical theory, asthma, eosinophil (Eosinophil, EOS) - infiltration of airway inflammation, but in recent years many studies have found that the oxidation / antioxidant imbalance also plays an important role in asthma development. Ambroxol (ambroxol, AMB) is a clinical common expectorant, research in recent years found that it has certain antioxidant effect and has certain application value in the treatment of asthma.
Objective To observe the effect of ambroxol on lung tissue of asthmatic rats and the pathological changes of the lung tissue of asthmatic rats and bronchoalveolar lavage fluid in activated protein -1 (activator protein-1 AP-1) and gamma glutamyl cysteine synthetase (y-glutamylcysteine, synthetase, y-GCS) effect of egg white, preliminary study the effects of ambroxol on airway oxidative / asthma rats the role of antioxidant system and its possible mechanism.
Methods 30 male Sprauge-Dawley (SD) rats were randomly divided into 3 groups: control group, asthma group and ambroxol treatment group. The asthma group and the ambroxol treatment group were 1D and 8D to 1ml rats by intraperitoneal injection of antigen liquid, containing ovalbumin ((ovalbumin, OVA) 100mg, aluminum hydroxide 100mg) sensitized. Fifteenth day 1%OVA inhalation (w/v) induced airway responses, 20 minutes each time, 1 times a day, a total of 7 days. The blank control group on day 1,8 intraperitoneal injection of saline 1ml, fifteenth day inhalation of saline, 20 minutes each time, 1 times a day, a total of 7 days.
The ambroxol treatment group given intraperitoneal injection of ambroxol sensitized 50mg.kg-1.d-1, blank control group and asthma model group were given normal saline intraperitoneal injection for 21 days. In each animal after the last inhalation 24 hours underwent bronchoalveolar lavage, collected bronchoalveolar lavage fluid (bronchoalveolar lavage, fluid, BALF) by HE staining. To observe the lung tissue pathological changes, immunohistochemical staining was detected in bronchoalveolar lavage fluid (AP-1 c-Jun and c-Fos subunits) and the expression of y-GCS and immunohistochemistry (AP-1 c-Jun and c-Fos subunit) and y-GCS expression in the lung tissue.
Results (1) during the excitation process, the respiratory distress symptoms (breathing fast, cyanosis, etc.) appeared in the asthma model group. The symptoms of the rats in the ambroxol treatment group were significantly reduced compared with those in the asthma model group, and no symptoms such as respiratory distress were found in the blank control group.
(2) the pathological changes of lung tissue of rats in each group: blank control group showed rat lung alveolar size uniform and thin alveolar wall, bronchial epithelial integrity, there are almost no infiltration of inflammatory cells and its secretion. The asthma model group showed bronchial epithelial edema accompanied by partial loss, basement membrane thickening, mucous gland increased. Each layer of the pipe wall are visible eosinophil infiltration of macrophages and monocytes. The ambroxol treatment group showed the lesion and degree were significantly reduced compared with model group;
(3) in lung tissue and bronchoalveolar lavage fluid (AP-1 c-Jun, c-Fos subunit) protein expression in asthma model group lung tissue and bronchoalveolar lavage and expression of blank c-Jun protein and c-Fos protein in BAL fluid increased significantly compared to the control group (P0.01); ambroxol treatment group of c-Jun protein and c-Fos protein expression levels and asthma the model group decreased significantly (P0.01), but still higher than the control group (P0.01);
(4) in lung tissue and bronchoalveolar lavage fluid gamma -GCS protein expression: the expression level and blank of asthma model group lung tissue and bronchoalveolar lavage fluid gamma -GCS protein increased significantly compared to the control group (P0.01); ambroxol treatment group lung tissue and bronchoalveolar lavage fluid in gamma -GCS protein expression level compared with the asthma model group (P0.01) significantly decreased, but still higher than the control group (P0.01).
Conclusion (1) ambroxol plays an antioxidant role, and its mechanism may be related to the reduction of the expression level of AP-1 (c-Jun, c-Fos subunit) and gamma -GCS protein.
(2) ambroxol reduces the damage of bronchopulmonary tissue through its antioxidation and has a certain protective effect on the model of asthmatic rats.

【学位授予单位】：新乡医学院
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R562.25

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