水通道蛋白對氣道高反應(yīng)大鼠呼吸道粘液分泌的影響
發(fā)布時(shí)間：2019-05-26 18:57
【摘要】：目的：氣道高反應(yīng)性(airway hyperresponsiveness, AHR)是指一些外來物質(zhì)或反應(yīng)原分子進(jìn)入氣道從而刺激氣道所產(chǎn)生的一些高度應(yīng)激狀態(tài)和反應(yīng)過程,實(shí)驗(yàn)內(nèi)容①利用外源刺激因子臭氧建立一個(gè)失穩(wěn)態(tài)的模型,同時(shí)利用一些相關(guān)理化指標(biāo)來確定該模型的成功建立；②觀察臭氧應(yīng)激后大鼠肺組織水通道蛋白(aquaporin, AQP)的表達(dá)情況,分析氣道高反應(yīng)過程中AQPs的異常表達(dá)；③觀察臭氧應(yīng)激后支氣管分泌的粘蛋白總量、肺組織粘蛋白基因MUC5AC表達(dá)改變和異常表達(dá)的AQPs變化,探討異常表達(dá)的AQPs對臭氧應(yīng)激大鼠氣道粘液分泌的量和質(zhì)影響。 方法：①選用20只雄性SD大鼠將其任意的分成2組：正常對照組；臭氧應(yīng)激組每組10只。分別用檢測一些指標(biāo)包括總蛋白含量測定、支氣管肺泡灌洗液中細(xì)胞的分類和數(shù)量、大鼠模型的呼氣相氣道阻力(expiratory airwayresistance, Re)以及氣道中支氣管的病理切片等來確定大鼠應(yīng)激模型的成功建立；②選取20只SD大鼠任意將其分為2組：正常對照組；臭氧應(yīng)激組每組10只。采用熒光實(shí)時(shí)定量RT-PCR和Western Blot檢測臭氧應(yīng)激后大鼠肺組織三種AQPs(AQP1、AQP4、AQP5)的表達(dá),分析氣道高反應(yīng)過程中,AQPs的異常表達(dá)；③30只SD大鼠任意將其分為3組每組10只：正常對照組；臭氧應(yīng)激組；臭氧應(yīng)激+地塞米松治療組；分組檢測支氣管分泌粘蛋白總量、肺組織粘蛋白基因MUC5AC表達(dá)改變和臭氧應(yīng)激后肺組織三種異常表達(dá)的AQPs(AQP1、AQP4、AQP5)的量,分析它們之間的相關(guān)性。 結(jié)果：①臭氧(ozone, O3)是諸多可損傷氣道的外源性刺激因子其中的一種,用其攻擊支氣管上皮細(xì)胞破壞其結(jié)構(gòu)完整性從而建立高氣道模型,并且通過檢測方法①中所提到的各類理化指標(biāo)有力的證實(shí)了氣道高反應(yīng)性的存在；②大鼠肺組織中存在著多種水通道蛋白如AQP1、AQP4、AQP5均有表達(dá)且AQP5的表達(dá)最多；臭氧應(yīng)激后的大鼠肺組織內(nèi)AQP1、AQP4、AQP5表達(dá)明顯下調(diào)；③大鼠肺組織有MUC5AC表達(dá),臭氧應(yīng)激數(shù)天后,大鼠氣道粘液分泌量明顯增加,且MUC5AC表達(dá)隨之增多,兩者呈現(xiàn)正相關(guān)趨勢。④隨著AQPs的表達(dá)下降,氣道高反應(yīng)大鼠粘液分泌量明顯增多,MUC5AC的表達(dá)也增多。提示AQPs的表達(dá)與氣道粘液分泌總量呈現(xiàn)負(fù)相關(guān)的趨勢；AQPs的表達(dá)與粘液中的MUC5AC表達(dá)呈現(xiàn)負(fù)相關(guān)的趨勢。 結(jié)論：由于氣道上皮細(xì)胞在臭氧的刺激下出現(xiàn)了功能缺陷或失穩(wěn)狀態(tài),從而導(dǎo)致了氣道高反應(yīng)的發(fā)生。在氣道高反應(yīng)性過程中,粘液高分泌狀態(tài)與MUC5AC的表達(dá)水平有關(guān),AQPs的表達(dá)下調(diào)可增加支氣管粘液的分泌量和MUC5AC的表達(dá)。我們分析認(rèn)為AQP1、AQP4、AQP5的降低可能導(dǎo)致了氣道高反應(yīng)時(shí)肺水在毛細(xì)血管與肺泡間的轉(zhuǎn)運(yùn)失衡,加重氣道炎癥、釋放多種炎癥介質(zhì),使纖毛上皮和杯狀細(xì)胞比例失衡,最終導(dǎo)致水清除障礙,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.
【學(xué)位授予單位】：湖南師范大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2012
【分類號】：R363
本文編號：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.
【學(xué)位授予單位】：湖南師范大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2012
【分類號】：R363
【參考文獻(xiàn)】
相關(guān)期刊論文 前5條
1 李翔;王笑梅;張堅(jiān)松;;兩種氣道高反應(yīng)大鼠模型的建立與比較[J];湖南師范大學(xué)學(xué)報(bào)(醫(yī)學(xué)版);2009年01期
2 吉寧飛;卞濤;陳力;符曉蘇;殷凱生;;支氣管哮喘大鼠模型的建立與氣道反應(yīng)性的測定[J];南京醫(yī)科大學(xué)學(xué)報(bào)(自然科學(xué)版);2006年11期
3 秦曉群;氣道上皮細(xì)胞的抗氧化損傷保護(hù)及微環(huán)境調(diào)控[J];生理科學(xué)進(jìn)展;1999年02期
4 秦曉群,向陽,管茶香,張長青,孫秀泓;整合素配體結(jié)合反應(yīng)上調(diào)兔支氣管上皮細(xì)胞抗氧化能力[J];生理學(xué)報(bào);2001年01期
5 李娟,張耀庭,曾偉,羅璇,廖長春;應(yīng)用考馬斯亮藍(lán)法測定總蛋白含量[J];中國生物制品學(xué)雜志;2000年02期
，本文編號：2485555
本文鏈接：http://sikaile.net/xiyixuelunwen/2485555.html
最近更新
教材專著
