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研究生:邱雅鈴
研究生(外文):Ya- Ling Chiou
論文名稱:探討Derp2對於大鼠氣管平滑肌初代細胞之刺激作用與Pentoxifylline對於血小板衍生生長因子引起細胞增生之抑制作用
論文名稱(外文):Study of Stimulatory Effects of Der p2 on Rat Airway Smooth Muscle Primary Cells and Inhibitory Effects of Pentoxifylline on the Cells Proliferation Induced by Platelet-Derived Growth Factor
指導教授:林清淵林清淵引用關係
指導教授(外文):Ching- Yuang Lin
學位類別:博士
校院名稱:國立陽明大學
系所名稱:微生物及免疫學研究所
學門:生命科學學門
學類:微生物學類
論文種類:學術論文
論文出版年:2009
畢業學年度:97
語文別:中文
論文頁數:145
中文關鍵詞:氣管平滑肌細胞塵蟎類鐸受器非選擇型環核酸磷酸二酯酶抑制劑
外文關鍵詞:ASMCsDer p2PentoxifyllineERK1/2Akt
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氣喘病理機制之一是氣管平滑肌細胞 (airway smooth muscle cells, ASMCs) 受到過敏原刺激,會引起細胞的活化、增生與發炎反應。在台灣,塵蟎中的 Der p2 是重要的過敏原,但它與平滑肌細胞的之間作用目前仍不清楚,因此,吾人以初代培養之大鼠氣管平滑肌細胞(ASMCs)為標的細胞 (target cells),探討 Der p2 對於ASMCs之刺激作用,並探討臨床用藥Pentoxifylline (PTX) 抑制ASMCs活化增生的機制。研究結果發現, Der p2可以與細胞上的第二類型類鐸受器(Toll-like receptor 2,TLR2)相互作用,使得較多的Myeloid differentiation factor (MyD)88與TLR2連結,進行下游的訊息傳遞,導致活化extracellular signal-regulated kinases 1/2 ( ERK1/2)和c-Fos的表現,進而刺激ASMCs產生發炎媒介物,包括第六介白素(interleukin-6; IL-6)和第一型單核球趨化蛋白(Monocyte chemoattractant protein-1; MCP-1),因而使得呼吸道呈現持續發炎反應,此結果證明Der p2可活化ASMCs及引起發炎性反應。ASMCs被過敏原刺激後的增生反應也是造成氣喘的主要病理機制之一,所以抑制ASMCs的增生是治療氣喘的重要目標。吾人以非選擇型環核酸磷酸二酯酶(cyclic nucleotide phosphodiesterase, PDE) 抑制劑—PTX探討抑制氣管平滑肌細胞增生以及其機制。吾人利用血小板衍生生長因子(Platelet-derived growth factor,PDGF) 刺激ASMCs後再接受PTX處理,實驗結果支持PTX是藉由抑制Akt/nuclear factor of kappa B (NF-κB)而不是藉由p70S6K的訊息傳導來達到抑制PDGF所引起的ASMCs增生,吾人的研究提供了PTX治療氣喘的可能機制。
Pathological characteristics of asthma include the activation, hyperplasia, and inflammation in airway smooth muscle cells (ASMCs) stimulated by allergens. In Taiwan, Der p2 of dust mite is a common allergen, but the interaction between Der p2 and smooth muscle cells is not yet clear. In this thesis, primary ASMCs from rat tracheal were used as target cells and effects of Der p2 on ASMCs were studied. The inhibitory mechanisms of pentoxifylline (PTX) on ASMCs proliferation induced by platelet-derived growth factor (PDGF) were investigated. The results indicated that Der p2 could interact with toll-like receptor 2 (TLR2) and increase the binding of myeloid differentiation factor (MyD) 88 to TLR2. This increased interaction further to activate the phosphorylation of extracellular signal-regulated kinases 1/2 (ERK1/2) and expression of c-Fos. Der p2 enhanced inflammatory cytokines containing interleukin-6 (IL-6) and monocyte chemoattractant protein-1 (MCP-1) productions in ASMCs. These results proved that Der p2 activated the TLR2 signaling pathway and caused an inflammatory response in ASMCs that might be related to Der p2-induced bronchial hyperresponsiveness. In the pathogenesis of asthma, the proliferation of ASMCs is a key factor in airway remodeling and blocking of ASMCs prolifation may be a potential therapeutic target. PTX is a phosphodiesterase (PDE) inhibitor that can inhibit platelet-derived growth factor (PDGF)-stimulated proliferation of ASMCs. The results of this study demonstrated that PTX suppressed the Akt/nuclear factor of kappa B (NF-κB) activation, not the p70S6K pathway, and then inhibited PDGF-induced cell proliferation, thereby revealing a possible role of PTX in the treatment of asthma.
目錄
縮寫對照表 3
中文摘要 5
英文摘要 6
第一章 緒論 7
第一節 氣喘的概論 8
一、氣喘的盛行 8
二、氣喘特徵 8
三、氣喘的成因 9
四、氣喘與環境的關係 12
第二節 氣喘與過敏原 15
一、過敏原的種類 15
二、家塵蟎(house dust mite allergen)與致敏化 17
三、Der p2參與氣喘形成的機制 18
第三節 氣喘與氣管平滑肌細胞 19
一、氣管平滑肌是氣道發炎的的主要角色之一 20
二、氣管平滑肌增生刺激劑 23
三、刺激劑刺激氣管平滑肌細胞增生的機制 24
四、氣管平滑肌細胞肥大 26
第四節 過敏原與氣管平滑肌作用 27
一、 類鐸受器(Toll like receptors, TLRs) 27
二、過敏原透過TLRs影響氣喘形成 29
三、過敏原透過TLRs影響氣管平滑肌的活化 31
第五節 以Phosphodiesterase inhibitors治療氣管平滑肌的增生 32
一、Phosphodiesterase 抑制劑的簡介 32
二、Pentoxifylline 的簡介 33
三、Pentoxifylline抑制細胞增生 35
第二章 材料與方法 36
第一節 實驗材料 37
第二節 實驗方法 46
第三章 實驗結果 55
第一部份 建立初代培養氣管平滑肌細胞 56
第二部份 家塵蟎的成分Der p2影響氣管平滑肌細胞的機制 58
第三部分 Pentoxifylline抑制氣管平滑肌細胞增生的機制 64
第四章 討論 68
參考文獻 71
圖表 83
附錄 127
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