線粒體損傷在支氣管哮喘發(fā)生機(jī)制中的作用
發(fā)布時(shí)間：2018-08-21 08:38
【摘要】：支氣管哮喘是一種復(fù)雜的炎癥疾病，它伴有各種程度的氣流受限，氣道高反應(yīng)性和氣道炎癥。它通常是由環(huán)境因素及炎癥刺激相互結(jié)合引起的。最近人們研究發(fā)現(xiàn)線粒體的功能和哮喘的發(fā)病機(jī)制有著密切的聯(lián)系，他們發(fā)現(xiàn)線粒體的功能紊亂在哮喘的氣道炎癥中起重要作用。那么在哮喘中線粒體會(huì)發(fā)生怎樣的變化呢？本次研究我們將通過哮喘動(dòng)物模型及對線粒體超微結(jié)構(gòu)的觀察來探討哮喘中線粒體結(jié)構(gòu)發(fā)生了哪些變化及這些變化在哮喘中的作用。線粒體是細(xì)胞內(nèi)的一種重要的細(xì)胞器，是細(xì)胞內(nèi)的能量發(fā)生器,可以產(chǎn)生機(jī)體需要的ATP。同時(shí)，線粒體也在其他許多生理過程中都扮演著重要角色,例如在體內(nèi)葡萄糖的代謝作用以及在細(xì)胞水平上影響胞內(nèi)的鈣信號、活性氧(ROS)的生成和細(xì)胞凋亡等。那么，線粒體對胞質(zhì)內(nèi)鈣信號和活性氧會(huì)有怎樣的調(diào)控作用呢？研究已表明生理狀態(tài)下，線粒體通過釋放和攝取的途徑對胞漿內(nèi)鈣離子濃度進(jìn)行調(diào)控，而胞漿的鈣穩(wěn)態(tài)對維持細(xì)胞內(nèi)的正常生理活動(dòng)有重要作用，當(dāng)外界炎癥刺激導(dǎo)致線粒體破壞時(shí)，細(xì)胞內(nèi)鈣穩(wěn)態(tài)會(huì)破壞，鈣離子濃度會(huì)產(chǎn)生波動(dòng)。本次研究我們通過測定炎癥刺激時(shí)鈣離子濃度的變化來探討鈣離子和炎癥的關(guān)系。同樣，線粒體的破壞也與活性氧的含量呈正相關(guān)，外界炎癥刺激細(xì)胞導(dǎo)致各種自由基的釋放，它們會(huì)損傷線粒體，導(dǎo)致線粒體超微結(jié)構(gòu)的改變，被破壞的線粒體會(huì)通過氧化磷酸化產(chǎn)生過量的活性氧，大量的活性氧會(huì)對細(xì)胞組織造成損傷，這對氣道上皮細(xì)胞的纖毛擺動(dòng)頻率會(huì)產(chǎn)生怎樣的影響呢？本次研究將通過測定不同濃度活性氧模擬物過氧化氫對細(xì)胞纖毛擺動(dòng)頻率的影響來反映活性氧與炎癥的關(guān)系。 目的 本次研究利用卵清蛋白來激發(fā)大鼠建立哮喘模型，并對大鼠的氣道反應(yīng)性和各項(xiàng)炎癥反應(yīng)進(jìn)行檢測；通過電鏡觀察氣道線粒體的結(jié)構(gòu)來說明線粒體在支氣管哮喘中的變化；通過測定炎癥刺激時(shí)氣道上皮細(xì)胞中活性氧及鈣離子濃度以及不同濃度活性氧對氣道上皮細(xì)胞纖毛擺動(dòng)頻率的影響來反映線粒體損傷在支氣管哮喘中的機(jī)制。 方法 1.用卵清蛋白來激發(fā)大鼠建立哮喘模型，利用呼吸機(jī)和多導(dǎo)生理信號記錄儀來測定大鼠氣道反應(yīng)性，利用共聚焦顯微鏡測定纖毛擺動(dòng)頻率，用瑞士染色涂片對嗜酸性粒細(xì)胞進(jìn)行計(jì)數(shù)，，用ELISA對血漿中炎癥因子進(jìn)行檢測，用HE染色分析分析大鼠的肺組織形態(tài)， 2.用電鏡來觀察線粒體超微結(jié)構(gòu)， 3.用IL-4刺激氣道上皮細(xì)胞并用共聚焦顯微鏡測定細(xì)胞內(nèi)的ROS及鈣離子濃度；用不同濃度的活性氧刺激細(xì)胞并測定纖毛擺動(dòng)頻率。 結(jié)果 1.與正常組大鼠相比，哮喘大鼠氣道反應(yīng)性明顯增高（P0.05）；氣道結(jié)構(gòu)發(fā)生重構(gòu)，氣管管壁明顯增厚（P0.05）,管壁周圍有大量炎癥細(xì)胞浸潤；氣管上皮纖毛擺動(dòng)頻率降低；肺泡灌洗液中嗜酸性粒細(xì)胞數(shù)量明顯增多；血漿中IL-8和TNF-的含量明顯增高（p0.05）；這些結(jié)果說明哮喘時(shí)大鼠機(jī)體發(fā)生了炎癥反應(yīng); 2.與正常組大鼠相比，哮喘組大鼠氣道內(nèi)線粒體的超微結(jié)構(gòu)發(fā)生改變，出現(xiàn)明顯的腫脹與破壞；說明哮喘時(shí)氣道內(nèi)線粒體結(jié)構(gòu)發(fā)生破壞，其導(dǎo)致的功能損傷在哮喘的發(fā)病機(jī)制起重要作用; 3.與正常組細(xì)胞相比，IL-4刺激的氣道上皮細(xì)胞其活性氧含量與細(xì)胞內(nèi)鈣離子濃度明顯增高（P0.05），其纖毛擺動(dòng)頻率明顯降低（P0.05）；用不同濃度H2O2刺激細(xì)胞時(shí)，細(xì)胞纖毛擺動(dòng)頻率隨濃度的增高而降低，均低于正常組（P0.05）。 結(jié)論 上述結(jié)果表明哮喘時(shí)線粒體的結(jié)構(gòu)會(huì)發(fā)生明顯的破壞，損傷后的線粒體會(huì)導(dǎo)致胞漿內(nèi)鈣離子濃度的升高及大量活性氧的釋放，活性氧的釋放會(huì)導(dǎo)致纖毛擺動(dòng)頻率的降低，使其無法維持正常的呼吸道功能，這將進(jìn)一步加重炎癥，導(dǎo)致哮喘遷延不愈。
[Abstract]:Bronchial asthma is a complex inflammatory disease accompanied by various degrees of airflow restriction, airway hyperresponsiveness and airway inflammation. It is usually caused by the combination of environmental factors and inflammatory stimuli. Recent studies have found that the function of mitochondria is closely related to the pathogenesis of asthma. They have found that the function of mitochondria is closely related to the pathogenesis of asthma. Disturbance plays an important role in airway inflammation in asthma. So what happens to mitochondria in asthma? In this study, we will explore the changes of mitochondrial structure in asthma and the role of these changes in asthma through asthma animal models and observation of mitochondrial ultrastructure. An important organelle is an intracellular energy generator that produces the ATP needed by the body. At the same time, mitochondria also play an important role in many other physiological processes, such as glucose metabolism in vivo, calcium signaling at the cellular level, production of reactive oxygen species (ROS) and cell apoptosis. How does mitochondria regulate intracytoplasmic calcium signaling and reactive oxygen species? Studies have shown that under physiological conditions, mitochondria regulate intracytoplasmic calcium concentration by means of release and uptake, and cytoplasmic calcium homeostasis plays an important role in maintaining normal cellular physiological activities, when inflammation stimulates mitochondria. In this study, we investigated the relationship between calcium ion and inflammation by measuring the changes of calcium ion concentration during inflammatory stimulation. Similarly, the destruction of mitochondria is positively correlated with the content of reactive oxygen species (ROS). External inflammation stimulates the release of various free radicals in cells, and they Mitochondria will be damaged, resulting in ultrastructural changes in mitochondria, the damaged mitochondria will produce excessive reactive oxygen species through oxidative phosphorylation, a large number of reactive oxygen species will cause damage to the cell tissue, which will affect the frequency of cilia swing in airway epithelial cells? The effect of hydrogen peroxide on cell cilia oscillation frequency reflects the relationship between reactive oxygen species and inflammation.
objective
In this study, ovalbumin was used to stimulate asthma model in rats, and the airway reactivity and inflammatory reactions were detected; the structure of airway mitochondria was observed by electron microscopy to explain the changes of mitochondria in bronchial asthma; the concentration of reactive oxygen species (ROS) and calcium ions in airway epithelial cells were measured by inflammatory stimulation. And the effect of different concentrations of reactive oxygen species on cilia oscillation frequency of airway epithelial cells to reflect the mechanism of mitochondrial damage in bronchial asthma.
Method
1. Establish asthma model in rats stimulated by ovalbumin, measure airway responsiveness by respirator and multi-channel physiological signal recorder, measure cilia swing frequency by confocal microscope, count eosinophils by Swiss staining smear, detect inflammatory factors in plasma by ELISA, and analyze by HE staining. Lung tissue morphology in rats
2. electron microscopy was used to observe the ultrastructure of mitochondria.
3. The airway epithelial cells were stimulated by IL-4 and the intracellular concentrations of ROS and Ca2+ were measured by confocal microscopy, and the ciliary oscillation frequencies were measured by reactive oxygen species with different concentrations.
Result
1. Compared with the normal group, the airway responsiveness of asthmatic rats was significantly increased (P 0.05); the airway structure was reconstructed, the tracheal tube wall was significantly thickened (P 0.05), there were a large number of inflammatory cells infiltrated around the tracheal wall; the frequency of ciliary oscillation of tracheal epithelium was decreased; the number of eosinophils in alveolar lavage fluid was significantly increased; the contents of IL-8 and TNF-in plasma were significantly increased. The amount was significantly higher (P0.05); these results indicated that inflammation occurred in asthmatic rats.
2. Compared with normal rats, the ultrastructure of airway mitochondria in asthmatic rats was changed, and there was obvious swelling and destruction.
3. Compared with the normal group, the content of reactive oxygen species (ROS) and intracellular calcium concentration of IL-4-stimulated airway epithelial cells were significantly increased (P 0.05), and the frequency of ciliary oscillation was significantly decreased (P 0.05).
conclusion
These results suggest that the structure of mitochondria in asthma may be destroyed obviously, and the damaged mitochondria will lead to the increase of calcium concentration and the release of a large number of reactive oxygen species in the cytoplasm. The release of reactive oxygen species will lead to the decrease of ciliary oscillation frequency, making it unable to maintain normal respiratory function, which will further aggravate inflammation and lead to asthma. Delay is not healed.
【學(xué)位授予單位】：華中科技大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2013
【分類號】：R562.25
[Abstract]:Bronchial asthma is a complex inflammatory disease accompanied by various degrees of airflow restriction, airway hyperresponsiveness and airway inflammation. It is usually caused by the combination of environmental factors and inflammatory stimuli. Recent studies have found that the function of mitochondria is closely related to the pathogenesis of asthma. They have found that the function of mitochondria is closely related to the pathogenesis of asthma. Disturbance plays an important role in airway inflammation in asthma. So what happens to mitochondria in asthma? In this study, we will explore the changes of mitochondrial structure in asthma and the role of these changes in asthma through asthma animal models and observation of mitochondrial ultrastructure. An important organelle is an intracellular energy generator that produces the ATP needed by the body. At the same time, mitochondria also play an important role in many other physiological processes, such as glucose metabolism in vivo, calcium signaling at the cellular level, production of reactive oxygen species (ROS) and cell apoptosis. How does mitochondria regulate intracytoplasmic calcium signaling and reactive oxygen species? Studies have shown that under physiological conditions, mitochondria regulate intracytoplasmic calcium concentration by means of release and uptake, and cytoplasmic calcium homeostasis plays an important role in maintaining normal cellular physiological activities, when inflammation stimulates mitochondria. In this study, we investigated the relationship between calcium ion and inflammation by measuring the changes of calcium ion concentration during inflammatory stimulation. Similarly, the destruction of mitochondria is positively correlated with the content of reactive oxygen species (ROS). External inflammation stimulates the release of various free radicals in cells, and they Mitochondria will be damaged, resulting in ultrastructural changes in mitochondria, the damaged mitochondria will produce excessive reactive oxygen species through oxidative phosphorylation, a large number of reactive oxygen species will cause damage to the cell tissue, which will affect the frequency of cilia swing in airway epithelial cells? The effect of hydrogen peroxide on cell cilia oscillation frequency reflects the relationship between reactive oxygen species and inflammation.
objective
In this study, ovalbumin was used to stimulate asthma model in rats, and the airway reactivity and inflammatory reactions were detected; the structure of airway mitochondria was observed by electron microscopy to explain the changes of mitochondria in bronchial asthma; the concentration of reactive oxygen species (ROS) and calcium ions in airway epithelial cells were measured by inflammatory stimulation. And the effect of different concentrations of reactive oxygen species on cilia oscillation frequency of airway epithelial cells to reflect the mechanism of mitochondrial damage in bronchial asthma.
Method
1. Establish asthma model in rats stimulated by ovalbumin, measure airway responsiveness by respirator and multi-channel physiological signal recorder, measure cilia swing frequency by confocal microscope, count eosinophils by Swiss staining smear, detect inflammatory factors in plasma by ELISA, and analyze by HE staining. Lung tissue morphology in rats
2. electron microscopy was used to observe the ultrastructure of mitochondria.
3. The airway epithelial cells were stimulated by IL-4 and the intracellular concentrations of ROS and Ca2+ were measured by confocal microscopy, and the ciliary oscillation frequencies were measured by reactive oxygen species with different concentrations.
Result
1. Compared with the normal group, the airway responsiveness of asthmatic rats was significantly increased (P 0.05); the airway structure was reconstructed, the tracheal tube wall was significantly thickened (P 0.05), there were a large number of inflammatory cells infiltrated around the tracheal wall; the frequency of ciliary oscillation of tracheal epithelium was decreased; the number of eosinophils in alveolar lavage fluid was significantly increased; the contents of IL-8 and TNF-in plasma were significantly increased. The amount was significantly higher (P0.05); these results indicated that inflammation occurred in asthmatic rats.
2. Compared with normal rats, the ultrastructure of airway mitochondria in asthmatic rats was changed, and there was obvious swelling and destruction.
3. Compared with the normal group, the content of reactive oxygen species (ROS) and intracellular calcium concentration of IL-4-stimulated airway epithelial cells were significantly increased (P 0.05), and the frequency of ciliary oscillation was significantly decreased (P 0.05).
conclusion
These results suggest that the structure of mitochondria in asthma may be destroyed obviously, and the damaged mitochondria will lead to the increase of calcium concentration and the release of a large number of reactive oxygen species in the cytoplasm. The release of reactive oxygen species will lead to the decrease of ciliary oscillation frequency, making it unable to maintain normal respiratory function, which will further aggravate inflammation and lead to asthma. Delay is not healed.
【學(xué)位授予單位】：華中科技大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2013
【分類號】：R562.25
【共引文獻(xiàn)】
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