臺灣博碩士論文加值系統

過敏性氣喘 (allergic asthma) 已成為近20年來國人十分重視的
慢性疾病之一，由過敏而導致的氣喘現象常造成患者呼吸困難，胸悶等現象，嚴重者有可能因呼吸道黏液阻塞或氣道緊縮而死亡。造成過敏性氣喘的原因由於是接吸入過敏原 (allergens)，如塵蟎，因而誘發上呼吸道過度反應(airway hyper-responsiveness)，簡稱AHR。到目前為止腐食酪蟎(Tyrophagus putrescentiae)，而尚未有動物模式的資料，因此本論文以Tyrophagus putrescentiae為過敏原探討在動物誘發呼吸道過度反應的機轉，建立未來研究減敏治療或藥物治療的基礎，本研究中已證實以腐食酪蟎粗萃所得到取蛋白質經過三次腹腔注射可使Balb/c小鼠IgE升高，達成致敏狀態，經過二次鼻腔滴入過敏原能誘發小鼠產生過敏性氣喘的，觀察處理組小鼠之肺功能的指標(Penh)有明顯的上升。然後將小鼠犧牲，藉由流式細胞儀螢光量偵測小鼠血液中T細胞所分泌兩種細胞激素IFN-γ和IL-4，發現有過敏性氣喘之小鼠可偵測到較高量的IL-4，證明血液中的T細胞偏向Th2類型。同時蒐集小鼠肺泡沖洗液(BALF)，去觀察氣管細胞的組成比例，分析結果發現有過敏性氣喘小鼠的肺泡沖洗液中，含有許多發炎細胞如嗜中性球和嗜酸性球，證實氣喘小鼠的肺內有發炎現象。再利用組織切片觀察肺部支氣管與肺部組織，經病理切片觀察上呼吸道的柱狀上皮細胞破壞和發炎細胞浸潤的現象，以上結果可初步證實腐食酪蟎在實驗動物誘發過敏性氣喘。本實驗室已有細菌表現出腐食酪蟎重组蛋白rTyr p3其大小約為27KDa。於是我們利用rTyr p3對過敏性氣喘小鼠進行減敏治試驗，發現有減緩AHR的現象。

The prevalence of allergic asthma has been doubled in the last 10-15 years, this leads to the disease becomes one of the important chronic diseases in Taiwan. Tyrophagus putrescentiae is one of common allergens in the environment, and can widely major cause of allergic athsma. In our study, we use crude extract of T. putrescentiae as an allegen to study the mechanism of pathogenesis of allergic asthma in a murine model. we attempted to use a murine model to study the mechanism of pathogenesis of allergic asthma caused by the crude extract of Tp. After sensitization, the level of IgE against Tp in sensitized animals increased. To test the lung function, the enhanced pause (Penh) was found to be higher in sensitized animals than that of control mice. After challenge, the cell proportion of neutrophil and eosinophil in bronchoalveolar lavage fluid (BALF) in the sensitized mice was higher than that of control group. Examination of histological sections showed that the bronchia and lung columnar epithelium of the sensitized mice was destroyed but not in control group. Moreover, flow cytometry also revealed that CD4+ cells especially Th2 type cells increased significantly in sensitized animals
According the results, we esrablished basic allergic asthmatic murine model that induced by Tyrophagus putrescentiae. It could used as a good model for some advanced researchs in therapy. Our lab first found a recombinant protein in T. putrescentiae called rTyr p3. The molecular weight is about 27KDa was identified as major allergen. 5B.The frequency of of 86% patients’ sera was detected to have IgE binding to rTyr p 3 by ELISA The localization of rTyr p3 was analyzed by immunofluorescent assay in T. putrescentiae. The major allergen rTyr p3 was localized in gut、mouthparts and faecal particles. In this study, the outcome of hyposensitization with rTyr p3 in asthmatic mice inicated the decrease of Penh .

中文摘要……………………………………………………… 4
英文摘要……………………………………………………… 6
壹、 緒論 ……………………………………………………8
貳、 材料與方法 …………………………………………… 15
參、 結果 …………………………………………………… 38
肆、 討論 …………………………………………………… 44
伍、 參考文獻 ……………………………………………… 57
陸、 圖 ……………………………………………………… 50
柒、 表 62
捌、 附錄 …………………………………………………… 64

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