臺灣博碩士論文加值系統

近年來研究發現胸腺基質淋巴生成素(Thymic stromal lymphopoietin, TSLP) 與過敏性發炎疾病包含過敏性皮膚炎、鼻炎和氣喘相關。目前文獻報告已指出TSLP在過敏及氣喘疾病中表現量會增加。TSLP的訊號傳遞起始接受器是透過TSLP接受器複合體(TSLPR complex)來進行，而TSLP接受器複合體具有二個次單元，分別為介白素-7接受器α鏈(IL-7 receptor-α (IL-7Rα) chain) 和TSLPR chain。本篇論文的探討主題為分析TSLPR chain及IL-7Rα chain與氣喘的關聯性。TSLPR chain會表現在樹突細胞、T細胞、B細胞、肥大細胞、自然殺手細胞、嗜伊紅性白血球和單核球細胞。而IL-7Rα chain則會表現在骨髓淋巴前趨細胞、pro-B細胞、自然殺手細胞、naïve T細胞、CD4+ T細胞、CD8+ T細胞、樹突細胞、單核球細胞、肥大細胞及嗜伊紅性白血球。會共同表現TSLPR chain及IL-7Rα chain的細胞有T細胞、單核球細胞、樹突細胞、自然殺手細胞、肥大細胞及嗜伊紅性白血球。而目前從文獻報告上發現先天遺傳因素會影響TSLPR chain和IL-7Rα chain的表現，報告指出TSLPR chain基因多型性可能與異位體質性氣喘的敏感性有關；而IL-7Rα chain基因多型性則與過敏性氣喘的敏感性有關。而也有文獻報告指出在過敏疾病中，TSLPR chain陽性細胞若存在於過敏發炎前趨激素及其它因子(例如:過敏原、細菌產物、脂質媒介物)的環境下會影響TSLPR chain的表現，此結果表示後天環境因素也可能會影響TSLPR chain的表現。目前從我們實驗數據發現在過敏性氣喘(atopic、asthmatic, AA)組小朋友周邊血液單核細胞 (Peripheral blood mononuclear cells, PBMC)中的TSLPR chain和IL-7Rα chain mRNA表現量確實較無過敏無氣喘( non-atopic、non-asthmatic, NN )組小朋友有顯著的增加。而在生物科技系葉怡玲老師實驗室學生洪煒等人的Der p 2刺激人類THP-1細胞株的實驗，結果發現Der p2刺激並不會影響TSLPR chain和IL-7Rα chain mRNA表現量。但在其它種類細胞上TSLPR complex調控的機制究竟是經由先天基因多型性的影響還是由後天環境因素刺激調控亦或是由先天加後天影響，這些都還是需要再深入探討。或許TSLPR complex的表現量可提供作為氣喘嚴重度指標。

Recent studies found that cytokine thymic stromal lymphopoietin (TSLP) play a very important role and related with allergic inflammatory diseases including atopic dermatitis, rhinitis, and asthma. The TSLP receptor (TSLPR) is a heterodimeric cytokine receptor consisting of the IL-7 receptor alpha chain (IL-7Rα) and a TSLP-specific receptor chain. In our research, our specific aim is focused on the relationship between thymic stromal lymphopoietin receptor complex expression and the severity pediatric asthma. TSLPR chain can express on dendritic cells, T cells, B cells, mast cells, natural killer (NK) cells, eosinophil cells and monocytes. IL-7Rα chain can express on bone marrow lymphoid precursors, pro-B cells, NK cells, naïve and memory CD4+ and CD8+ T cells, dendritic cells, monocytes/macrophages, mast cells and eosinophil cells. T cells, monocytes, dendritic cells, NK cells, mast cells and eosinophil cells can coexpress the TSLPR chain and IL-7Rα chain. In some report, they found that genetic polymorphisms will affect the expression of the TSLPR chain and IL-7Rα chain. The SNP of TSLPR chain is one possible factor correlated with the susceptibility to atopic asthmatic disease. Also, polymorphisms of IL-7Rα chain are associated with allergic asthma susceptibility. One report found that TSLPR expressing cells have potential to modulate expression of the TSLPR when exist in a milieu of proallergic and proinflammatory cytokines and other factors (e.g., allergens, bacterial products, and lipid mediators). This result indicates acquired environmental factors can also affect the expression of the TSLPR. Until now, it is unclear that if the expression of TSLPR complex is correlated with genetic factors or environmental factors or both. In recent study, we found that TSLPR chain and IL-7Rα chain mRNA expression in PBMC layer of the AA (atopic, asthmatic) group children is more significantly higher than NN (non-atopic, non-asthmatic) group children. In another study of Hung et al., stimulation of human THP-1 cell by Der p2 did not affect the expression of TSLPR chain and IL-7Rα chain. The TSLP receptor expression mechanism in other cell types is still need further investigation. We hope that the expression of TSLPR can be applied to the evaluation of the severity of asthma and to be as a disease marker in the future.

中文摘要...................................................i
英文摘要.................................................iii
致謝.......................................................v
目錄......................................................vi
表目錄....................................................ix
圖目錄.....................................................x
縮寫對照表...............................................xii
第一章 文獻探討............................................1
1.1氣喘簡介................................................1
1.2過敏簡介................................................2
1.3過敏反應致病機轉........................................2
1.4 胸腺基質淋巴生成素(Thymic stromal lymphopoietin,TSLP)..4
1.5 胸腺基質淋巴生成素接受器複合體(Thymic stromal
lymphopoietin receptor complex,TSLPR complex)簡介......6
1.6 文獻指出影響TSLPR chain及IL-7Rα chain表現的因素.......9
1.7 國內外專利檢索........................................11
第二章 研究目的...........................................12
第三章 材料與方法.........................................13
3.1 本次臨床實驗的人類血液檢體來源及組別分類方式..........13
3.2 血液檢體處理..........................................13
3.3 周邊血液單核細胞(PBMC)分離...........................14
3.4 RNA純化..............................................14
3.5 分光光度計測RNA含量...................................15
3.6 反轉錄-聚合酶連鎖反應Reverse Transcription-Polymerase
Chain Reaction (RT-PCR)...............................15
3.6.1 Reverse Transcription (RT).....................15
3.6.2 PCR............................................15
3.7 電泳..................................................16
3.7.1 1.2% agarose gel配製方式.......................17
3.7.2 膠片染色.......................................17
3.8 Integrated Density Value(IDV)紀錄....................17
3.9 數據分析.............................................17
第四章 實驗結果...........................................18
4.1 臨床實驗所測得的結果..................................18
4.1.1藉由氣喘症狀發病與否分成無氣喘症狀組(NN + AN)及有氣
喘症狀組(NA + AA) 兩大族群，比較其TSLPR及IL-7Rα..18
4.1.2藉由過敏體質的有無分成無過敏體質組(NN+NA) 及過敏體質
組(AN+AA) 兩大族群，比較其TSLPR及IL-7Rα..........18
4.1.3再將無氣喘症狀及有氣喘症狀這兩大族群分別再以過敏體質
做比較，所以又得到NN 比AN組以及NA比AA組..........19
4.1.4四組檢體TSLPR chain的IDV表現結果比較.............20
4.1.5四組檢體TSLPR與GAPDH相除後得到一個標準化的值(％)，
結果比較.........................................20
4.1.6四組檢體IL-7Rα chain的IDV表現結果比較............21
4.1.7四組檢體IL-7Rα與GAPDH相除後得到一個標準化的值(%)，
結果比較..........................................21
第五章 討論...............................................23
第六章 結論...............................................29
參考文獻..................................................30
附表......................................................36
附圖......................................................37
英文論文大綱..............................................50
作者簡歷..................................................56

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