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研究生:鄭芳怡
研究生(外文):Jang Fang-Yi
論文名稱:探討肺纖維母細胞及Toll-likereceptor在氣喘動物模式中所扮演的角色
論文名稱(外文):The role of fibroblast and Toll-like receptor in asthma animal model
指導教授:葉怡玲葉怡玲引用關係
指導教授(外文):Ye Yi-Ling
學位類別:碩士
校院名稱:中華醫事學院
系所名稱:生物科技研究所
學門:生命科學學門
學類:生物科技學類
論文種類:學術論文
論文出版年:2007
畢業學年度:94
語文別:中文
論文頁數:80
外文關鍵詞:Dermatophagoide peteronyssinus groups 2FibroblastNerve growth factorThymic Stromal lymphoietinand Toll-like receptor
相關次數:
  • 被引用被引用:1
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  • 下載下載:141
  • 收藏至我的研究室書目清單書目收藏:2
中文摘要
過敏性氣喘是一種呼吸道發炎疾病,當過敏原與肺組織接觸,先天性免疫反應被啟動,接著導致發炎細胞聚集與趨化激素分泌,最後導致Th2 免疫發展。肺組織最主要的4種細胞類型,包含上皮細胞、纖維母細胞、巨噬細胞及內皮細胞。為了研究先天性免疫機制在氣喘疾病,我們利用氣管內注入過敏原方式於C57BL/6小鼠,連續7日,然後評估肺泡沖洗液內的浸潤細胞。我們發現在ovalbumin (OVA)及基因重組製造的 Dermatophagoide peteronyssinus group 2蛋白 (rDer p2) 刺激後,小鼠嗜酸性白血球與淋巴球分別有增加的現象,此結果指出我們以短期刺激肺上皮組織可以造成呼吸道發炎現象。進一步研究肺纖維母細胞在過敏性致敏期的角色,我們利用初代肺纖維母細胞經OVA或rDer p2刺激後,結果發現會分泌大量RANTES、Eotaxin、TNF-a及TGF-b。有趣地,於rDer p2或OVA刺激組中亦可偵測到thymic stromal cell-derived lymphopoietin (TSLP)及nerve growth factor (NGF)的分泌量有增加現象。最後,我們利用中和試驗與C3H/HeJ小鼠來證明toll-like receptor (TLR) 2 及4是否參與了這個反應。結果發現在anti-TLR2 及anti-TLR4 抗體的作用下TNF-a分泌量有劑量相對受抑制的現象。而TLR4缺陷鼠C3H/HeJ的 TNF-a分泌表現量比正常鼠C3H/HeN低,並達到顯著差異。由此得知,TLR2及TLR4皆有參與Der p2刺激纖維母細胞分泌細胞激素的作用。分析纖維母細胞在氣喘致敏期的角色將對於先天性免疫反應在氣喘疾病的機制有更清楚的了解並希望對於疾病治療有所貢獻。
Abstract
Allergic asthma is an inflammatory disease of bronchial airways. When allergens contact with lung tissue, the innate immune response was initiated, subsequently, leaded to inflammatory cells accumulation and chemokine secretion, and finally resulted in Th2 immune development. There are four major cell types in lung tissue including epithelial cells, fibroblasts, macrophages, and endothelial cells. In order to study the mechanism of innate immunity in the asthma disease, we intratrachealy administrated allergens into C57BL/6 mice for seven days then to evaluate the infiltrated cells in bronchial alveolar lavage fluids (BALF). We found that eosinophils and lymphocytes respectively accumulated in ovalbumin (OVA)- and recombinant Dermatophagoide peteronyssinus (rDer p2)-treated mice, indicating that lung epithelium shortly stimulated by allergens caused to airway inflammation. Furthermore, primary lung fibroblasts stimulated with OVA or rDer p2 secreted higher levels of RANTES, eotaxin, TNF-a, and TGF-b. Interestingly, elevated levels of the thymic stromal cell-derived lymphopoietin (TSLP) and nerve growth factor (NGF) also detected in OVA- or rDer p2-treated groups. Finally, we use neutralizing assay to evaluate whether the toll-like receptor (TLR) involved in this phenomena. We found that using anti-TLR2 and anti-TLR4 antibodies could dose-dependently inhibited TNF-a secretion by rDer p2 stimulation, moreover. C3H/HeJ mice secreted significantly lower level of TNF-a compared to C3H/HeN mice. Summarizing the above data, TLR2 and TLR4 both involved in the cytokine secretion by rDer p2 stimulation. To study the role of lung fibroblasts on the airway sensitizing stage will contribute to more clear realization of the innate immune response in asthma disease.
目錄
標題
口試委員會審定書
授權書
中文摘要
英文摘要
誌謝
縮寫表
目錄
表目錄
圖目錄
第一章 緒論
一. 過敏性氣喘介紹
二. 過敏性氣喘定義
三. 過敏性氣喘致病機轉
四. Toll-like receptor在氣喘疾病中所扮演的角色
五. 過敏原
六. 肺纖維母細胞在氣喘疾病中所扮演的角色
七. 趨化激素
第二章 論文研究目的
第三章 實驗材料與方法
1.實驗動物
2.過敏原致敏小鼠模式建立
3.小鼠肺纖維母細胞初代培養
4.流式細胞儀分析肺細胞分群
5. recombinant Der p2純化與分析
6.利用ELISA分析趨化激素與細胞激素分泌量
7.利用RT-PCR方式偵測TLR的基因表現
8.小鼠肺纖維母細胞TLR中和試驗
第四章 實驗結果
第五章 討論
第六章 圖表
參考文獻
附錄
附錄ㄧ 實驗儀器及耗材
附錄二 實驗藥品來源
附錄三 常用藥品配置方法
附錄四 核酸引子序列及最適作用溫度
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