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研究生:白惠珍
研究生(外文):Hui-ChenPai
論文名稱:Thrombin刺激肺部纖維母細胞產生結締組織生長因子經JAK2及STAT3之訊息角色探討
論文名稱(外文):The Roles of JAK2 and STAT3 in Thrombin-Induced Connective Tissue Growth Factor Expression in Human Pulmonary Fibroblasts
指導教授:林建煌林建煌引用關係
學位類別:碩士
校院名稱:臺北醫學大學
系所名稱:醫學研究所
學門:醫藥衛生學門
學類:醫學學類
論文種類:學術論文
論文出版年:2007
畢業學年度:95
語文別:中文
論文頁數:63
中文關鍵詞:肺纖維化、結締組織生長因子
外文關鍵詞:lung fibroblasts、CTGF、JAK2、STAT3
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在本篇論文中,我們將探討thrombin調控纖維母細胞connective tissue growth factor (CTGF)的表現之詳細分子機轉。在WI-38纖維母細胞中,thrombin可誘導CTGF蛋白表現及CTGF-luciferase的活性。給予AG490 (JAK抑制劑)及JAK DN可依劑量反應關係,抑制thrombin誘導CTGF蛋白的表現及CTGF-luciferase的活性。 Thrombin以時間相關性地誘導JAK2之 Tyr1007/1008 的磷酸化。細胞給予轉染STAT3 DN及 STAT ODN (oligodeoxynucleotides)抑制thrombin誘導CTGF蛋白的表現及CTGF-luciferase的活性。 Thrombin誘導STAT 之Tyr705的磷酸化呈現時間相關性,並且藉由DNA-binding affinity pull down assay證實thrombin可以誘導STAT結合至CTGF基因起始區上。Thrombin誘導STAT3的磷酸化可被AG490及JAK DN所抑制。我們也發現thrombin誘導CTGF蛋白表現也可被兩種STAT3結合序列的ODN (STAT3-1 ODN及STAT3-2 ODN)所抑制。再者,thrombin所誘導CTGF蛋白表現及CTGF-luciferase的活性,可被c-Src DN抑制。Thrombin誘導STAT3磷酸化及JAK2磷酸化也可被c-Src DN抑制。綜合以上的結果推測thrombin可經由c-Src,JAK2及STAT3訊息路徑來誘導WI-38細胞之CTGF蛋白的表現。
In this study, we investigated the signaling pathway by thrombin- induced CTGF expression. Thrombin induced increase in CTGF protein expression and CTGF-luciferase activity in WI-38 lung fibroblasts. AG490 (JAK inhibitor) and dominant negative of JAK2 (JAK2 DN) inhibited thrombin-CTGF expression and CTGF-luciferase activity in a dose-dependent manner. Thrombin also caused a time-dependent increase JAK2 phosphorylation at Tyr1007/1008. Transfection of cells with STAT3 DN and STAT3 ODN (oligodeoxynucleotides) inhibited CTGF protein expression and CTGF-luciferase activity. Thrombin induced STAT3 at Tyr705 phosphorylation in a time-dependent manner, which was inhibited by AG490 and JAK2 DN. Thrombin induced STAT binding to CTGF promoter by DNA-binding affinity pull down assay. We also found that thrombin-induced CTGF protein expression was inhibited by STAT3-1 oligodeoxynucleotides(ODN) and STAT3-2 (ODN). Furthermore, thrombin-indused CTGF protein expression and CTGF-luciferase activity was inhibited by cell transfected with c-Src DN. Thrombin induced STAT3 phosphorylation and JAK2 phosphorylation also inhibited by c-Src DN. Taken together, thrombin though Src, JAK2, and STAT3 signaling pathway to induced CTGF protein expression and activity in lung fibroblasts.
目錄
壹、 中文摘要………………………………..………………………1
貳、 英文摘要…………………………………….………………..2-3
參、 緒論………………………………………………...………..4-12
肆、 實驗材料與方法…………………………………………...13-18
一.實驗材料……………………………………………………..…13
二.實驗方法…………………………………………….………14-18
伍、 結果…………………………………………………….......19-24
一. JAK參與 thrombin刺激WI-38纖維母細胞CTGF蛋白的表現…………………………………………………………...19-20
二. Thrombin 經由活化STAT3的活化來誘導CTGF的表現…………………………………………………………...20-22
三. STAT Decoy oligodeoxynucleotides(ODN)降低thrombin誘導CTGF蛋白的表現……………………………………...….22-23
四. Thrombin經由活化 c-Src 誘導 CTGF 的表現………………………………………………………...…23-24
五. Thrombin經由c-Src來活化JAK2及STAT3來誘導CTGF的表現………………………………………………………………24
陸、 討論.......................................................................................25-30
柒、 參考文獻…………………………………………………...31-45
捌、 圖表………………………………………………………...46-62
玖、 附圖……………………………………………………………63
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