本文關(guān)鍵詞： 氨溴索 支氣管哮喘 激活蛋白-1 γ-谷胺酰半胱氨酸合成酶　出處：《新鄉(xiāng)醫(yī)學院》2012年碩士論文　論文類型：學位論文

【摘要】：背景經(jīng)典理論認為,哮喘是以嗜酸粒細胞(Eosinophil,EOS)浸潤為主的氣道炎癥,但近年來很多研究發(fā)現(xiàn),氧化／抗氧化失衡在哮喘的發(fā)生發(fā)展中也起著重要作用。氨溴索(ambroxol,AMB)是一種臨床常用祛痰藥,近年來研究發(fā)現(xiàn),其具有一定抗氧化作用,在哮喘治療中具有一定的應用價值。 目的通過觀察氨溴索對哮喘大鼠肺組織病理的改變以及對哮喘大鼠肺組織及支氣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)蛋白的影響,初步探討氨溴索對哮喘大鼠氣道氧化/抗氧化系統(tǒng)的作用及其可能的作用機制。 方法將30只雄性Sprauge-Dawley(SD)大鼠隨機分為3組：空白對照組、哮喘模型組和氨溴索處理組。哮喘模型組和氨溴索處理組分別于第1d和第8d向大鼠腹腔內(nèi)注射抗原液1ml,(含有卵清蛋白(ovalbumin,OVA)100mg,氫氧化鋁100mg)致敏。第15天開始霧化吸入1%OVA (w/v)激發(fā)氣道反應,每次20分鐘,每日1次,共7天�？瞻讓φ战M則于第1、8天腹腔內(nèi)注射生理鹽水1ml,第15天開始霧化吸入生理鹽水,每次20分鐘,每日1次,共7天。 氨溴索處理組致敏同時給予腹腔注射氨溴索50mg.kg-1.d-1,空白對照組和哮喘模型組則同時給予等量生理鹽水腹腔注射,共21天。各組動物于最后一次霧化吸入后24小時行支氣管肺泡灌洗,收集支氣管肺泡灌洗液(bronchoalveolar lavage fluid, BALF)。采用HE染色觀察肺組織病理的改變,免疫細胞化學染色法檢測支氣管肺泡灌洗液中AP-1(c-Jun和c-Fos亞單位)及y-GCS的表達水平以及免疫組織化學法觀察AP-1(c-Jun和c-Fos亞單位)及y-GCS在肺組織中的表達。 結(jié)果(1)激發(fā)過程中哮喘模型組大鼠出現(xiàn)呼吸窘迫癥狀(呼吸急促、青紫等),氨溴索處理組大鼠激發(fā)的癥狀與哮喘模型組相比有明顯減輕,空白對照組大鼠未見呼吸窘迫等癥狀。 (2)各組大鼠肺組織病理變化：空白對照組顯示大鼠肺組織可見肺泡大小均一且肺泡壁薄,支氣管上皮完整,周圍幾乎無炎性細胞浸潤及其分泌物。哮喘模型組顯示支氣管粘膜上皮水腫伴有局部的脫落,基底膜的增厚,粘液腺增多,管壁的各層均可見嗜酸性粒細胞、巨噬細胞和單核細胞的浸潤。氨溴索處理組顯示病變范圍、程度均較模型組明顯減輕； (3)肺組織和支氣管肺泡灌洗液AP-1(c-Jun、c-Fos亞單位)蛋白表達：哮喘模型組肺組織及支氣管肺泡灌洗液中c-Jun蛋白和c-Fos蛋白的表達水平與空白對照組比較明顯升高(P0.01)；氨溴索處理組c-Jun蛋白和c-Fos蛋白的表達水平與哮喘模型組比較明顯降低(P0.01),但仍高于空白對照組(P0.01)； (4)肺組織和支氣管肺泡灌洗液γ-GCS蛋白表達：哮喘模型組肺組織及支氣管肺泡灌洗液γ-GCS蛋白的表達水平與空白對照組比較明顯升高(P0.01)；氨溴索處理組肺組織及支氣管肺泡灌洗液中γ-GCS蛋白的表達水平與哮喘模型組比較明顯降低(P0.01),但仍高于空白對照組(P0.01)。 結(jié)論(1)氨溴索發(fā)揮抗氧化作用,其機制可能是通過降低AP-1(c-Jun、c-Fos亞單位)和γ-GCS蛋白的表達水平有關(guān)。 (2)氨溴索通過其抗氧化作用,減輕支氣管肺組織的損傷,對哮喘大鼠模型具有一定的氣道保護作用。
[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.

【學位授予單位】：新鄉(xiāng)醫(yī)學院
【學位級別】：碩士
【學位授予年份】：2012
【分類號】：R562.25

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