本文選題：PM + 氣道炎癥��； 參考：《浙江大學》2017年碩士論文

【摘要】：近年來,由于工業(yè)發(fā)展以及城市化進程的推進,環(huán)境污染越來越嚴重,越來越多的國內外研究表明,空氣中一定濃度的顆粒物,對于一些呼吸系統(tǒng)疾病包括哮喘、COPD、肺癌以及心血管系統(tǒng)疾病的發(fā)病率的上升有著重要的影響。PM主要包括工業(yè)生產排放、自然源中土壤灰塵、機動車尾氣或是生物燃料及礦物燃料燃燒所產生,PM顆粒物進入氣道會通過堆積在氣道或者通過支氣管進入肺泡,從而能夠引起許多呼吸系統(tǒng)及肺部疾病或者加重原有的氣道炎癥疾病。PM的危害與其顆粒物的大小有著必然的聯系顆粒物的顆粒直徑越小時,顆粒物的表面積就會越大而且其表面所附著的有毒物質也會增加,同時顆粒進入氣道后的沉降速度也會變慢,同時進入肺泡的機會也會變大,對肺部的危害也會更嚴重。研究發(fā)現暴露于一定濃度顆粒物的環(huán)境時可誘導明顯的氣道高反應性,也會誘導以中性粒細胞為主的氣道炎癥;可加重哮喘氣道炎癥,從而導致哮喘患者對激素治療敏感性降低,一定濃度顆粒物處理細胞也會增強氣道上皮細胞內氧化應激水平從而誘導氣道上皮間質的轉化。目前有很多研究將誘導炎癥細胞死亡而清除浸潤的炎癥細胞作為治療炎癥性疾病的一個重要的治療方法。而且有研究表明凋亡抑制蛋白Bcl-2的抑制劑能夠通過誘導嗜酸性粒細胞或者中性粒細胞的凋亡從而比較有效的治療呼吸氣道炎癥例如哮喘特別是對于激素治療抵抗的中性粒細胞型哮喘。本研究在浙江大學呼吸疾病研究所關于PM課題研究的基礎上去探討PM誘導的氣道炎癥是否與凋亡抑制蛋白Bcl-2有關系,并探討凋亡抑制蛋白Bcl-2的抑制劑是否能夠緩解PM誘導氣道炎癥從而開發(fā)治療PM誘導氣道炎癥的新策略。目的:體內研究建立PM誘導氣道炎癥模型,研究凋亡抑制蛋白Bcl-2在PM誘導的氣道炎癥中的作用,并利用Bcl-2過表達小鼠去驗證是否凋亡抑制蛋白在PM誘導的氣道炎癥中發(fā)揮重要作用同時利用Bcl-2抑制劑去處理體內PM誘導的氣道炎癥模型,探究其是否能夠緩解此類炎癥。方法:體外首先用標準化的PM氣道滴注WT小鼠,建立氣道炎癥模型,流式細胞術運用AnnexiV/PI檢測炎癥細胞的凋亡,探究是否PM誘導的氣道炎癥與炎癥細胞的凋亡有關。同時檢測PM干預后小鼠肺泡灌洗液細胞中凋亡抑制蛋白Bcl-2的表達,探討是否炎癥細胞的凋亡與Bcl-2相關。然后利用Vav-Bcl-2轉基因小鼠氣道滴注PM,炎癥Bcl-2在PM誘導的氣道炎癥中的作用。最后利用Bcl-2選擇性抑制劑ABT199處理PM干預后的小鼠,看是否能夠給緩解PM引起的氣道炎癥,并利用Annexi V/PI檢測炎癥細胞的凋亡從而確定ABT199是否是通過增加炎癥細胞的凋亡從而緩解PM引起的氣道炎癥。結果:連續(xù)兩天氣道滴注PM會引起肺泡灌洗液總數、肺泡灌洗液中中性粒細胞比例明顯增加,病理切片結果也也說明PM能夠明顯增加炎癥細胞的聚集。Annexi V/PI檢測炎癥細胞的凋亡結果說明PM干預后炎癥細胞的凋亡與NS組相比明顯增加。且IF及流式檢測結果也表明BALF細胞中凋亡抑制蛋白Bcl-2表達也有明顯增加。Bcl-2過表達小鼠在PM氣道滴注后與WT小鼠相比,中性粒細胞數目、氣道炎癥的聚集及中性粒細胞趨化因子的表達都明顯增加而且炎癥自行減退的時間及程度都減慢。構建模型時在末次滴注PM之后24小時滴注Bcl-2選擇性抑制劑,結果發(fā)現Bcl-2抑制劑能夠明顯誘導中性粒細胞的凋亡從而減輕PM引起的氣道炎癥。結論:PM能夠誘導以中性粒細胞為主的氣道炎癥,并且這種氣道炎癥在Bcl-2過表達的小鼠體內會明顯加重且自行清理的時間延長,Bcl-2選擇性抑制劑ABT-199能夠通過誘導中性粒細胞的凋亡即炎癥細胞的凋亡從而減輕PM引起的氣道炎癥。
[Abstract]:In recent years, due to the development of industry and the advancement of urbanization, environmental pollution is becoming more and more serious. More and more domestic and foreign studies have shown that certain concentrations of particulate matter in the air have an important impact on the rise of some respiratory diseases including asthma, COPD, lung cancer and cardiovascular disease..PM mainly includes work. Production of industrial emissions, natural sources of soil dust, motor vehicle exhaust, or combustion of biofuels and fossil fuels, and PM particles entering the airway by accumulating in the airway or through the bronchus into the alveoli, which can cause many respiratory and pulmonary diseases or the damage to the original airway inflammation,.PM, and its grain. The larger the particle size is, the larger the particle size, the larger the surface area of the particles and the increase of the toxic substances attached to the surface, and the sedimentation rate will slow down after the particles enter the airway, and the chances of entering the alveoli will also become larger, and the damage to the lungs will be more serious. When exposed to a certain concentration of particulate matter, it can induce obvious airway hyperresponsiveness, and also induce airway inflammation dominated by neutrophils, which can aggravate airway inflammation in asthmatic patients, resulting in a decrease in sensitivity to hormone therapy in asthmatic patients. Certain concentration of granular substance treatment cells also enhance the level of oxidative stress within the airway epithelial cells. There are many studies that will induce the death of inflammatory cells and remove infiltrating inflammatory cells as an important treatment for the treatment of inflammatory diseases. Moreover, studies have shown that inhibitors of apoptosis inhibitory protein Bcl-2 can induce apoptosis of eosinophils or neutrophils by inducing apoptosis from eosinophilic granulocytes or neutrophils It is more effective to treat respiratory airway inflammation, such as asthma, especially for neutrophilic granulocytic asthma, which is resistant to hormone therapy. This study is based on the study of PM in the respiratory disease Research Institute of Zhejiang University to investigate whether PM induced airway inflammation is associated with apoptosis suppressor Bcl-2 and to explore the apoptosis suppressor protein Bcl-2 Whether or not the inhibitors can relieve the PM induced airway inflammation and develop a new strategy for the treatment of airway inflammation by PM. Objective: to establish a PM induced airway inflammation model in vivo, to study the role of apoptosis inhibitory protein Bcl-2 in the airway inflammation induced by PM, and to verify whether the apoptosis inhibitory protein is induced by PM in the PM induced by the Bcl-2 overexpressed rat. The airway inflammation plays an important role and the Bcl-2 inhibitor is used to deal with the PM induced airway inflammation model in vivo, and to explore whether it can relieve this kind of inflammation. Methods: in vitro, WT mice were injected with standardized PM airway in vitro to establish the airway inflammation model. Flow cytometry was used to detect the apoptosis of inflammatory cells by AnnexiV/PI, and to explore if PM was used. The induced airway inflammation is related to the apoptosis of inflammatory cells. At the same time, the expression of apoptosis suppressor protein Bcl-2 in the alveolar lavage cells of mice after PM was detected, and the apoptosis of the inflammatory cells was related to the Bcl-2. Then, PM was injected into the airway of the Vav-Bcl-2 transgenic mice, and the effect of inflammatory Bcl-2 in the airway inflammation induced by PM was used. Finally, the effect of PM on the airway inflammation induced by PM was used. Bcl-2 selective inhibitor ABT199 treatment of PM dry mice, see whether it can relieve airway inflammation caused by PM, and use Annexi V/PI to detect the apoptosis of inflammatory cells to determine whether ABT199 is by increasing the apoptosis of inflammatory cells to alleviate airway inflammation caused by PM. Fruit: two days of continuous infusion of PM in the airway will cause alveolar irrigation. The total number of lotion and the proportion of neutrophils in the alveolar lavage fluid increased obviously. The pathological section also showed that PM could significantly increase the aggregation of inflammatory cells.Annexi V/PI to detect the apoptotic results of inflammatory cells, indicating that the apoptosis of inflammatory cells increased significantly after PM intervention. And IF and flow detection results also indicated that BALF cells were withered in BALF cells. The expression of Bcl-2 also significantly increased the number of neutrophils, the aggregation of neutrophils and the expression of neutrophil chemotactic factor in the.Bcl-2 overexpressed mice compared with the WT mice, and the time and degree of inflammation self-reduction slowed down. 24 hours after the final infusion of PM, 24 hours after the final infusion of the model was constructed. Bcl-2 selective inhibitors were instilled. The results showed that Bcl-2 inhibitors could induce apoptosis of neutrophils and reduce airway inflammation caused by PM. Conclusion: PM can induce airway inflammation based on neutrophil, and this airway inflammation can be significantly increased in Bcl-2 overexpressed mice and the time of self-cleaning is prolonged, Bc ABT-199, a selective inhibitor of L-2, can alleviate airway inflammation caused by PM by inducing apoptosis of neutrophils, namely apoptosis of inflammatory cells.

【學位授予單位】：浙江大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R56
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本文編號：1820124