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研究生:許淑芬
研究生(外文):Shu-Fen Hsu
論文名稱:Galectin-1與Liver X Receptor 在神經細胞中之保護及分化作用之潛在影響
論文名稱(外文):Potential Protective And DifferentiativeEffects Of Galectin-1 And Liver X Receptor In Neural Cells
指導教授:江明璋王淑音王淑音引用關係
指導教授(外文):Ming-Chang ChiangShu-Yin Wang
口試委員:江明璋王淑音謝建正林恆
口試委員(外文):Ming-Chang ChiangShu-Yin WangChien-Cheng HsiehHeng Len
口試日期:2012-06-22
學位類別:碩士
校院名稱:中國文化大學
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:2012
畢業學年度:100
語文別:中文
論文頁數:60
中文關鍵詞:半乳糖凝集蛋白-1肝X受體
外文關鍵詞:Galectin-1Liver X Receptor
相關次數:
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近期有研究指出氧化型態之半乳糖凝集蛋白-1(GAL-1)及活化肝X受體(LXRs)在中樞神經系統及神經細胞中具減緩發炎反應之效果。此外,也有學者指出GAL-1 及LXRs與幹細胞之增生和分化作用息息相關。在本篇研究中我們利用過度表現之氧化型態GAL-1及LXR活化劑(TO901317)進一步探討對於N2A cells之神經保護機制以及對於人類神經幹細胞(hNSCs)之影響。在我們的研究結果顯示分別經由利用給予GAL1和LXR活化劑處理作用之下,其在N2A cells中具顯著促進神經細胞之軸突生長以及抑制腫瘤壞死因子α(TNF-α)所誘導之毒性的效果。而GAL1和LXR活化劑主要為可有效地減少腫瘤壞死因子α所誘導之核因子-NF-κB(NF-κB)的表現和及顯著地抑制NF-B易位進入細胞核內表現,在其作用機制中我們證明GAL-1和LXR活化劑可經由透過阻斷NF-κB活化減少細胞發炎因子如環氧化酶(COX-2)、誘導型一氧化氮合成酶(iNOS)之表現進一步降低發炎反應。此外,我們也發現GAL-1和LXR活化劑在人類神經幹細胞中之作用其可增加MAP2(神經元細胞標誌)之表現。經由我們的研究結果証明GAL-1和LXR活化劑在N2A cells之神經保護作用中確實扮演著重要的角色並且具有誘導人類神經幹細胞分化為神經元細胞之潛力。


Recent investigations suggest that oxidized Galectin-1 (GAL-1) and activation of liver X receptors ( LXRs ) attenuate the inflammatory response of neural cells. The current studies further characterize the mechanism of neural protection of GAL- 1and LXR agonist in N2A cells and human neural stem cells (hNSCs). The results showed that GAL1 and LXR agonist (TO901317) significantly promoted neurite outgrowth and suppressed tumor necrosis factor-α(TNF-α) induced toxicity. The current studies also showed that GAL1 and LXR agonist treatment could inhibit TNF-α induced nuclear factor-κB(NF-κB) expression and translocation into nucleus. In addition, we have been demonstrated that GAL-1 and LXR agonist could reduce proinflammatory cytokines such as cyclo-oxygenase-2 (COX-2) and inductible Nitric Oxide Synthase (iNOS) expression via blocking NF-B activation. These results suggest that the protective role of GAL-1 and LXR agonist in N2A cells was mediated through the suppression of NF-κB activation. Moreover, GAL-1 and LXR agonist also induced the expression of microtubule associated protein 2 (MAP2) in hNSCs. These results suggest that GAL-1 and LXR agonist plays important role neuroprotection in N2A cells and induces in hNSCs differentiation into neuronal cells.


致謝
Abstract……………………………………………………………………………………………………………………………………I
中文摘要……………………………………………………………………………II
圖、表目錄…………………………………………………………………………III
第一章 緒論……………………………………………………………………………1
第一節 細胞行為之調控因子與生物訊息傳遞機制………………………………1
第二節 Galectin1 ………………………………………………………………………………………………………………1
第三節 LXRs (Liver X receptors)……………………………………………………………………………2
第四節 NF-κB (Nuclear factor-κB)在發炎反應中所扮演之角色………2
第五節 Human neural Stem cell………………………………………………………………………………4
第六節 研究目的與動機……………………………………………………………6
第二章 文獻回顧………………………………………………………………………7
第三章 實驗材料與方法………………………………………………………………9
第一節 細胞培養與處理……………………………………………………………9
第二節 觀察細胞型態及計算突觸外生……………………………………………11
第三節 細胞存活率試驗...………………………………………………………12
第四節 細胞免疫螢光染色...……………………………………………………12
第五節 蛋白質定量…………………………………………………………………13
第六節 西方墨點法…………………………………………………………………13
第七節 定量聚合酶連鎖反應………………………………………………………14
第四章實驗結果………………………………………………………………………17
第一節 觀察給予GAL-1 及LXR活化劑對於N2A cells 之突觸外生(Neurite outgrowth)
之變化……………………………………………………………………………………17
第二節 探討N2A cells 在有血清及無血清之培養基中經由TNF-α的刺激作用下,給予
GAL-1 及LXR活化劑處理對於細胞存活率(Cell viability)之影響……………17
第三節 探討N2A cells 在有血清及無血清之培養基中經由TNF-α的刺激作用下,給予
GAL-1 及LXR活化劑處理對於所誘導NF-κB表現之影響……………………………18
第四節 探討N2A cells 在含有血清及無血清之培養基中,給予GAL-1及LXR活化劑處
理72小時後,經由TNF-α的刺激作用下對於所誘導NF-κB目標基因iNOS及 COX2
表現之影響………………………………………………………………………………18
第五節 探討GAL-1及LXR活化劑在hNSCs對於增殖或分化作用之影響……………………19
第五章 討論…………………………………………………………………………………………21
第一節 給予GAL-1及LXR活化劑處理後對於N2A cells 突觸生長之情形………………21
第二節 N2A cells 在含有血清及無血清之培養基中給予GAL-1及LXR活化劑處理後對於
TNF-α的刺激逆境反應中之細胞存活率之變化………………………………………21
第三節 N2A cells 在含有血清及無血清之培養基中,給予 GAL-1及LXR 活化劑處理72
小時後,經由TNF-α的刺激作用下對於所誘導NF-κB位移及其目標基因iNOS及COX2
表現之變化………………………………………………………………………………22
第四節 利用GAL-1及LXR活化劑調節hNSCs分化作用…………………………………………22
第六章 未來展望………………………………………………………………………………………24
第七章 圖、表說明……………………………………………………………………………………25
第八章 參考資料………………………………………………………………………………………57






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