臺灣博碩士論文加值系統

肺上皮細胞在氣喘及COPD上皆扮演一重要的角色。在TNF-α及IL-17等發炎激素刺激下，其會分泌出許多細胞激素(IL-6、IL-8)及介質(eotaxin、RANTES)來吸引發炎細胞的聚集以共同造成肺的發炎反應。IL-10一開始被定義為Th 2的細胞激素，後來發現其具有抗發炎之作用，可抑制發炎細胞激素IL-6和IL-8產生。而在我們之前的研究結果也證實IL-10 在治療氣喘發作小鼠上有不錯的療效，因此我們想更進一步要去了解IL-10在對TNF-α、IL-4、IL-13及IL-17所造成的肺上皮細胞之發炎時的抑制效果以及其調控的機制為何。首先我們建立起培養小鼠肺上皮細胞的平台，初步評估IL-4和TNF-α對小鼠肺上皮細胞誘發發炎反應的效果，觀察到在給予不同劑量的IL-4、TNF-α和不同作用時間下刺激肺上皮細胞，可促其分別表現發炎介質eotaxin 和IL-6；而在IL-10抑制發炎實驗結果，証實IL-10可有效抑制eotaxin 和IL-6的生成。之後更進一步用人的肺上皮細胞株A549 和BEAS-2B 為平台建立起在給予細胞激素(TNF-α、IL-4、IL-13和IL-17)刺激下，可誘發IL-6、IL-8、eotaxin和RANTES的表現。由結果得知：IL-10的確具有降低細胞表現IL-6、IL-8和RANTES的能力，且其抑制作用的大小與是否會誘發高量的IL-10R無關。我們觀察到IL-10主要可藉由減少TNF-α刺激A549細胞所誘發之IκBα的降解來降低IL-6和IL-8在mRNA及蛋白質的表現;此外IL-10也可以藉由抑制IL-17所誘發MAPK pathway中磷酸化ERK1/2蛋白的表現來阻斷IL-6和IL-8mRNA及蛋白質的生成，以達共同抗發炎的作用。綜合以上結果可得知：IL-10的確可藉由抑制各別的細胞活化訊號傳遞以及降低發炎激素IL-6與IL-8在mRNA及蛋白質上的表現，來達到對抗TNF-α和IL-17對表皮細胞所造成的發炎反應。

Bronchial epithelial cells play an important role in the asthma and COPD disease. Under the stimulation of pro-inflammatory cytokine (TNF-α, IL-17), lung epithelial cells produce many types of cytokines and chemokines, and many contribute to lung inflammation by recruiting inflammatory cells. IL-10 originally had been described as a Th2-associated cytokine. However, further studies revealed that IL-10 was a cytokine with potent anti-inflammatory properties and it could repress the expression of inflammatory cytokines such as IL-6 and IL-8. Our previous studies had demonstrated that IL-10 might have a therapeutic effect on animal model of asthma. Therefore, we wanted to investigate whether IL-10 plays an anti-inflammatory role on airway inflammation induced by TNF-α、IL-4、IL-13 and IL-17. Our preliminary data showed that TNF-α and IL-4 could stimulate mouse lung epithelium to secret IL-6 and eotaxin, respectively. Furthermore, we observed that IL-10 could significantly suppress IL-6 and eotaxin production from lung epithelium. In addition, human pulmonary epithelial cells A549 and BEAS-2B were used to investigate the expression of inflammatory mediators induced by TNF-α, IL-4, IL-13 and IL-17. The results showed that IL-10 could reduce the production of IL-6, IL-8 and RANTES from A549 cells. IL-10 had no effect on induction of IL-10 receptor expression. However, the mRNA expression of IL-6 and IL-8 was reduced by IL-10 treatment. In addition, TNF-α activated the NF-κB pathway and pretreatment of A549 cells with IL-10 reversed the effect of TNF-α on IκBα degradation. IL-10 also could attenuate IL-17- induced-IL-6 and IL-8 production by reducing the phosphorylation of ERK1/2 in MAPK pathway. In conclusion, we suggest that IL-10 markedly inhibits TNF-α- and IL-17-induced airway inflammation on A549 cells by decreasing IL-6 and IL-8 production.

謝誌..................... .................................I
目錄..................... ...............................III
中文摘要..................... ............................IV
英文摘要........................ ..........................V
第壹章 緒論.................... ..........................1
第一節 研究背景介紹......... ...........................2
1. 氣喘(asthma)與慢性阻塞性肺部疾病(chronic obstructive pulmonary disease)簡介喘(asthma)與慢性阻塞性肺部疾病(chronic obstructive pulmonary disease)簡介..........2
2. 氣喘致病機轉.................. ......................3
3. 慢性阻塞性肺部疾病致病機轉.... ......................5
4. 調控氣喘疾病................... .....................6
5. 調控慢性阻塞性肺部疾病.......... ....................7
6. 支氣管表皮細胞扮演角色............ ..................8
7. TNF-α及IL-17對呼吸道發炎反應的調控.................10
8. IL-10細胞激素對呼吸道發炎反應的調控 ................11
第貳章 研究動機與目的................... ................13
第参章 探討IL-10調控IL-4、IL-13與TNF-α所誘發小鼠肺表皮細胞之發炎反應.................... .................16
第一節 實驗目的與原理................... ..............17
第二節 實驗材料與方法.................... .............18
第三節 實驗結果........................... ............21
第肆章 探討IL-10如何調控IL-4、IL-13、TNF-α與IL-17所誘發人類肺表皮細胞之發炎反應..........................23
第一節 實驗目的與原理..................................24
第二節 實驗材料與方法..................................25
第三節 實驗結果........................................32
第五章 討論..............................................37
第一節 刺激肺表皮細胞誘發發炎反應......................38
第二節 IL-10調控細胞激素及發炎介質生成............... .41
第三節IL-10調控細胞內訊息傳遞蛋白表現.................43
第陸章 結論與未來方向..................................44
第柒章 圖表............................................46
第捌章 參考文獻........................... ..............70

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