水通道蛋白对气道高反应大鼠呼吸道粘液分泌的影响
发布时间：2019-05-26 18:57
【摘要】：目的：气道高反应性(airway hyperresponsiveness, AHR)是指一些外来物质或反应原分子进入气道从而刺激气道所产生的一些高度应激状态和反应过程,实验内容①利用外源刺激因子臭氧建立一个失稳态的模型,同时利用一些相关理化指标来确定该模型的成功建立；②观察臭氧应激后大鼠肺组织水通道蛋白(aquaporin, AQP)的表达情况,分析气道高反应过程中AQPs的异常表达；③观察臭氧应激后支气管分泌的粘蛋白总量、肺组织粘蛋白基因MUC5AC表达改变和异常表达的AQPs变化,探讨异常表达的AQPs对臭氧应激大鼠气道粘液分泌的量和质影响。 方法：①选用20只雄性SD大鼠将其任意的分成2组：正常对照组；臭氧应激组每组10只。分别用检测一些指标包括总蛋白含量测定、支气管肺泡灌洗液中细胞的分类和数量、大鼠模型的呼气相气道阻力(expiratory airwayresistance, Re)以及气道中支气管的病理切片等来确定大鼠应激模型的成功建立；②选取20只SD大鼠任意将其分为2组：正常对照组；臭氧应激组每组10只。采用荧光实时定量RT-PCR和Western Blot检测臭氧应激后大鼠肺组织三种AQPs(AQP1、AQP4、AQP5)的表达,分析气道高反应过程中,AQPs的异常表达；③30只SD大鼠任意将其分为3组每组10只：正常对照组；臭氧应激组；臭氧应激+地塞米松治疗组；分组检测支气管分泌粘蛋白总量、肺组织粘蛋白基因MUC5AC表达改变和臭氧应激后肺组织三种异常表达的AQPs(AQP1、AQP4、AQP5)的量,分析它们之间的相关性。 结果：①臭氧(ozone, O3)是诸多可损伤气道的外源性刺激因子其中的一种,用其攻击支气管上皮细胞破坏其结构完整性从而建立高气道模型,并且通过检测方法①中所提到的各类理化指标有力的证实了气道高反应性的存在；②大鼠肺组织中存在着多种水通道蛋白如AQP1、AQP4、AQP5均有表达且AQP5的表达最多；臭氧应激后的大鼠肺组织内AQP1、AQP4、AQP5表达明显下调；③大鼠肺组织有MUC5AC表达,臭氧应激数天后,大鼠气道粘液分泌量明显增加,且MUC5AC表达随之增多,两者呈现正相关趋势。④随着AQPs的表达下降,气道高反应大鼠粘液分泌量明显增多,MUC5AC的表达也增多。提示AQPs的表达与气道粘液分泌总量呈现负相关的趋势；AQPs的表达与粘液中的MUC5AC表达呈现负相关的趋势。 结论：由于气道上皮细胞在臭氧的刺激下出现了功能缺陷或失稳状态,从而导致了气道高反应的发生。在气道高反应性过程中,粘液高分泌状态与MUC5AC的表达水平有关,AQPs的表达下调可增加支气管粘液的分泌量和MUC5AC的表达。我们分析认为AQP1、AQP4、AQP5的降低可能导致了气道高反应时肺水在毛细血管与肺泡间的转运失衡,加重气道炎症、释放多种炎症介质,使纤毛上皮和杯状细胞比例失衡,最终导致水清除障碍,粘液高分泌。
[Abstract]:Objective: airway hyperreactive (airway hyperresponsiveness, AHR) refers to some high stress states and reaction processes produced by some foreign substances or reactive promolecules entering the airway to stimulate the airway. In content 1, an unstable model was established by using ozone, a foreign stimulator, and some related physical and chemical indexes were used to determine the successful establishment of the model. (2) to observe the expression of aquaporin (aquaporin, AQP) in lung tissue of rats after ozone stress, and to analyze the abnormal expression of AQPs in airway hyperresponsiveness. (3) to observe the total amount of mucin secreted by bronchus, the expression of mucin gene MUC5AC and AQPs in lung tissue after ozone stress, and to explore the effect of abnormal expression of AQPs on airway mucus secretion in ozone stress rats. Methods: 1 Twenty male SD rats were randomly divided into two groups: normal control group (n = 10) and ozone stress group (n = 10). Some indexes were measured, including total protein content, classification and quantity of cells in bronchoalveolar lavage fluid, and respiratory airway resistance (expiratory airwayresistance, in rat model. Re) and pathological sections of bronchus in airway were used to determine the successful establishment of stress model in rats. 2 20 SD rats were randomly divided into two groups: normal control group (n = 10) and ozone stress group (n = 10). The expression of three kinds of AQPs (AQP1,AQP4,AQP5) in lung tissue of rats after ozone stress was detected by fluorescence real-time quantitative RT-PCR and Western Blot, and the abnormal expression of AQPs in airway hyperresponsiveness was analyzed. There were 10 rats in each group: normal control group, ozone stress dexamethasone treatment group and ozone stress dexamethasone treatment group. The total amount of mucin secreted by bronchus, the expression of mucin gene MUC5AC in lung tissue and the amount of AQPs (AQP1,AQP4,AQP5) abnormal expression in lung tissue after ozone stress were detected, and the correlation between them was analyzed. Results: 1 Ozone (ozone, O 3) is one of many exogenous stimulators that can damage airway. It is used to attack bronchial epithelial cells and destroy its structural integrity to establish high airway model. The existence of airway hyperresponsiveness was confirmed by all kinds of physical and chemical indexes mentioned in method 1. (2) A variety of aquaporins, such as AQP1,AQP4,AQP5, were expressed in the lung tissue of rats, and the expression of AQP5 was the highest, while the expression of AQP1,AQP4,AQP5 in the lung tissue of rats after ozone stress was significantly down-regulated. 3The expression of MUC5AC was found in lung tissue of rats. After ozone stress, the secretion of airway mucus increased significantly, and the expression of MUC5AC increased, which showed a positive correlation trend. 4 with the decrease of AQPs expression, The mucus secretion and the expression of MUC5AC were significantly increased in rats with airway hyperresponsiveness. It is suggested that the expression of AQPs is negatively correlated with the total secretion of airway mucus, and the expression of AQPs is negatively correlated with the expression of MUC5AC in mucus. Conclusion: the airway hyperreaction occurs because of the functional defect or instability of airway epithelial cells stimulated by ozone. In the process of airway hyperresponsiveness, the hypersecretion of mucus is related to the expression of MUC5AC. The down-regulation of AQPs expression can increase the secretion of bronchial mucus and the expression of MUC5AC. Our analysis suggests that the decrease of AQP1,AQP4,AQP5 may lead to the imbalance of lung water transport between capillaries and alveoli during airway hyperreaction, aggravate airway inflammation, release a variety of inflammatory mediators, and make the proportion of ciliated epithelial and goblet cells unbalanced. Eventually, it leads to water clearance and high secretion of mucus.
【学位授予单位】：湖南师范大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R363
本文编号：2485555
[Abstract]:Objective: airway hyperreactive (airway hyperresponsiveness, AHR) refers to some high stress states and reaction processes produced by some foreign substances or reactive promolecules entering the airway to stimulate the airway. In content 1, an unstable model was established by using ozone, a foreign stimulator, and some related physical and chemical indexes were used to determine the successful establishment of the model. (2) to observe the expression of aquaporin (aquaporin, AQP) in lung tissue of rats after ozone stress, and to analyze the abnormal expression of AQPs in airway hyperresponsiveness. (3) to observe the total amount of mucin secreted by bronchus, the expression of mucin gene MUC5AC and AQPs in lung tissue after ozone stress, and to explore the effect of abnormal expression of AQPs on airway mucus secretion in ozone stress rats. Methods: 1 Twenty male SD rats were randomly divided into two groups: normal control group (n = 10) and ozone stress group (n = 10). Some indexes were measured, including total protein content, classification and quantity of cells in bronchoalveolar lavage fluid, and respiratory airway resistance (expiratory airwayresistance, in rat model. Re) and pathological sections of bronchus in airway were used to determine the successful establishment of stress model in rats. 2 20 SD rats were randomly divided into two groups: normal control group (n = 10) and ozone stress group (n = 10). The expression of three kinds of AQPs (AQP1,AQP4,AQP5) in lung tissue of rats after ozone stress was detected by fluorescence real-time quantitative RT-PCR and Western Blot, and the abnormal expression of AQPs in airway hyperresponsiveness was analyzed. There were 10 rats in each group: normal control group, ozone stress dexamethasone treatment group and ozone stress dexamethasone treatment group. The total amount of mucin secreted by bronchus, the expression of mucin gene MUC5AC in lung tissue and the amount of AQPs (AQP1,AQP4,AQP5) abnormal expression in lung tissue after ozone stress were detected, and the correlation between them was analyzed. Results: 1 Ozone (ozone, O 3) is one of many exogenous stimulators that can damage airway. It is used to attack bronchial epithelial cells and destroy its structural integrity to establish high airway model. The existence of airway hyperresponsiveness was confirmed by all kinds of physical and chemical indexes mentioned in method 1. (2) A variety of aquaporins, such as AQP1,AQP4,AQP5, were expressed in the lung tissue of rats, and the expression of AQP5 was the highest, while the expression of AQP1,AQP4,AQP5 in the lung tissue of rats after ozone stress was significantly down-regulated. 3The expression of MUC5AC was found in lung tissue of rats. After ozone stress, the secretion of airway mucus increased significantly, and the expression of MUC5AC increased, which showed a positive correlation trend. 4 with the decrease of AQPs expression, The mucus secretion and the expression of MUC5AC were significantly increased in rats with airway hyperresponsiveness. It is suggested that the expression of AQPs is negatively correlated with the total secretion of airway mucus, and the expression of AQPs is negatively correlated with the expression of MUC5AC in mucus. Conclusion: the airway hyperreaction occurs because of the functional defect or instability of airway epithelial cells stimulated by ozone. In the process of airway hyperresponsiveness, the hypersecretion of mucus is related to the expression of MUC5AC. The down-regulation of AQPs expression can increase the secretion of bronchial mucus and the expression of MUC5AC. Our analysis suggests that the decrease of AQP1,AQP4,AQP5 may lead to the imbalance of lung water transport between capillaries and alveoli during airway hyperreaction, aggravate airway inflammation, release a variety of inflammatory mediators, and make the proportion of ciliated epithelial and goblet cells unbalanced. Eventually, it leads to water clearance and high secretion of mucus.
【学位授予单位】：湖南师范大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R363
【参考文献】
相关期刊论文 前5条
1 李翔;王笑梅;张坚松;;两种气道高反应大鼠模型的建立与比较[J];湖南师范大学学报(医学版);2009年01期
2 吉宁飞;卞涛;陈力;符晓苏;殷凯生;;支气管哮喘大鼠模型的建立与气道反应性的测定[J];南京医科大学学报(自然科学版);2006年11期
3 秦晓群;气道上皮细胞的抗氧化损伤保护及微环境调控[J];生理科学进展;1999年02期
4 秦晓群,向阳,管茶香,张长青,孙秀泓;整合素配体结合反应上调兔支气管上皮细胞抗氧化能力[J];生理学报;2001年01期
5 李娟,张耀庭,曾伟,罗璇,廖长春;应用考马斯亮蓝法测定总蛋白含量[J];中国生物制品学杂志;2000年02期
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