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  本文選題：哮喘 + 納米粒子　； 參考：《南昌大學》2006年碩士論文

【摘要】：目的：建立屋塵螨哮喘動物模型，比較分別用屋塵螨粗浸液、重組Der p2、包被rDer p2納米粒子進行免疫治療的療效，為變態(tài)反應性疾病的免疫治療提供新的思路。 方法：首先利用PLGA(聚乳酸-羥乙酸)制備包被重組Der p2的納米粒子。其次，進行實驗動物分組：近交系清潔級6～8周齡BALB／c小鼠40只，體重18～20克，隨機分配為：A陰性對照組，B陽性對照組，C粗浸液治療組(簡稱：粗浸液組)，D rDer p2治療組(簡稱：rDer p2組)，E包裹rDer p2納米粒子治療組(簡稱：納米粒子組)，每組8只，雌雄各半。哮喘模型建立及干預措施：第0、7、14天腹膜內(nèi)注射(腹腔注射，i．p．)0.1mg／ml屋塵螨變應原粗浸液與10mg／ml Al(OH)_3生理鹽水的混懸液100μl：第28～30天對C、D、E組分別給予i．p．1mg／ml屋塵螨粗浸液100μl、1mg／ml rDer p2溶液100μl、1mg／ml包被rDer p2納米粒子溶液100 μl進行治療；第44～49天B、C、D、E組給予5mg／ml屋塵螨粗浸液100μl鼻腔滴入(i．n．)方式激發(fā)，A組一直相應給予生理鹽水處理；第51天處死各組動物。以酶聯(lián)免疫吸附試驗(ELISA)的方法檢測各組實驗動物血清中IgE、IgGl、IgG2a的水平，以及支氣管肺泡灌洗液(BALF)與脾細胞培養(yǎng)后上清液(SCCS)中細胞因子IL-4、IFN-γ及IL-10水平，并計數(shù)BALF中細胞總數(shù)與分類細胞數(shù)，制備肺組織病理切片以觀察其病理改變，并選擇視野用顯微攝像儀拍照。 結(jié)果：添加鋁佐劑的屋塵螨變應原致敏的小鼠在特異性變應原的連續(xù)激發(fā)下，肺組織產(chǎn)生明顯的炎癥反應，表現(xiàn)為BALF中細胞總數(shù)增加以及炎癥細胞所占比例及絕對計數(shù)升高，且以分泌Th2型細胞因子如IL-4為主。血清IgE、IgGl水平升高。肺組織病理切片觀察到支氣管、細支氣管上皮下和小血管周圍有明顯的炎性細胞浸潤，以嗜酸性粒細胞為主，，氣道上皮結(jié)構(gòu)紊亂，有部分脫落，部分可見粘液栓，上皮下充血水腫明顯，肺泡間質(zhì)增厚，也可見到
[Abstract]:Objective: to establish the animal model of house dust mite asthma and compare the efficacy of immunotherapy with the crude extract of house dust mite, recombinant Der p2 and rDer p2 nanoparticles, respectively, in order to provide a new idea for immunotherapy of allergic diseases. Methods: firstly, poly (lactic acid-glycolic acid) nanoparticles coated with recombinant Der p2 were prepared. Secondly, the experimental animals were divided into four groups: 40 inbred BALB/c mice of clean grade 6 and 8 weeks old, weighing 1820 grams. They were randomly divided into two groups: the control group with negative weight A and B positive control group with crude C crude extract group (abbreviated as the crude extract group) and the D rDer p2 treatment group (abbreviated as the rDer p2 group), which was coated with rDer p2 nanoparticles (8 rats in each group, half male and half female). Establishment of asthma model and intervention: intraperitoneal injection (i.p.lg / ml) intraperitoneal injection (i.p.) of 0.1 mg / ml crude extract of house dust mite allergen and 100 渭 l of 10mg/ml Al(OH)_3 normal saline solution on day 0, 714: 100 渭 lg / ml rDer p2 solution 100 渭 lg / ml, respectively, on day 2830, in group C + DX E RDer p2 nanoparticles were coated with 100 渭 l solution. On the 44th and 49th day, the rats in group E were given 100 渭 l crude extract of 5mg/ml house dust mites into their nasal cavity. Methods Group A was treated with normal saline and animals were killed on the 51st day. Elisa was used to detect the level of IgG2a in serum of experimental animals, and the levels of cytokines IL-4, IFN- 緯 and IL-10 in the supernatant of bronchoalveolar lavage fluid (BALF) and splenocytes. The total number of cells and the number of classified cells in BALF were counted, the pathological sections of lung tissue were prepared to observe the pathological changes, and the field of vision was photographed with a microscopic camera. Results: the mice sensitized with aluminum adjuvant caused by specific allergen produced obvious inflammatory reaction in lung tissue, which was manifested by the increase of the total number of cells, the proportion of inflammatory cells and the absolute count of inflammatory cells in BALF. And mainly secreted Th2 type cytokines such as IL-4. The level of IgGl in serum was increased. The inflammatory cell infiltration was observed in bronchus, bronchioles subcutaneously and around small vessels, mainly eosinophilic granulocytes. The airway epithelial structure was disordered, some of them fell off, and some mucus suppositories were observed. Epigastric hyperemia and edema, alveolar interstitial thickening, can also be seen
【學位授予單位】：南昌大學
【學位級別】：碩士
【學位授予年份】：2006
【分類號】：R392

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本文編號：1966803