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題名 雙酚合成物抑制氧化壓力及加強神經生長因子誘導神經突生長
The novel biphenol compounds inhibit oxidative stress and enhance nerve growth factor (NGF)-induced neurite outgrowth
作者 林芊瑜
貢獻者 詹銘煥
林芊瑜
關鍵詞 雙酚化合物
腎上腺髓質嗜鉻細胞瘤
神經保護
神經突生長
細胞外信號調節激酶(Erk1/2)
訊息傳遞轉錄活化基因-3(STAT3)
日期 2015
上傳時間 1-Sep-2015 16:18:16 (UTC+8)
摘要 人類隨著年齡增長後中樞神經系統的修補及再生能力逐漸下降，一旦神經系統受到傷害，是很嚴重的問題。因此，引導或促進神經細胞生長甚至再生的方法，中樞神經受損患者將獲得更有效的治療。先前已有文獻指出由植物厚朴萃取的天然化合物─和厚朴酚，具有抗氧化、抗腫瘤、抗發炎、神經保護與滋養的作用。在不同疾病模式的囓齒動物實驗，如帕金森氏症、阿茲海默症、癌症與腦缺血疾病等，和厚朴酚皆具有預防疾病或減緩症狀的效果。本篇研究使用和厚朴酚之衍生物─新合成雙酚化合物(MH102、MH103、MH104、MH106、MH107與MH111)，並探討對於神經細胞的保護與滋養作用。透過腎上腺髓質嗜鉻細胞瘤 PC12 細胞預先處理新合成雙酚化合物，並以過氧化氫(H2O2)使細胞產生氧化壓力，使用活性氧檢測試驗(DCFH-DA assay)偵測細胞內活性氧(reactive oxygen species, ROS)的含量。實驗結果顯示，預先處理較高濃度(3-10μM)的新合成雙酚化合物顯著降低過氧化氫所產生的氧化壓力。另以H2O2誘導PC12細胞死亡，並使用MTT試驗法，觀測新合成雙酚化合物對於細胞存活的影響。結果顯示新合成雙酚化合物顯著減少H2O2造成的細胞死亡。於神經滋養實驗，發現新合成雙酚化合物無法直接誘導PC12細胞的神經突生長。因此，使用神經滋養因子(nerve growth factor, NGF)誘導PC12細胞神經突生長，發現新合成雙酚化合物在低濃度(0.1-0.3μM)顯著加強神經突生長。然而雙酚化合物加強NGF誘導神經突生長之機制，並非透過活化細胞外信號調節激酶 (extracellular-signal-regulated kinases, Erk1/2)與訊息傳遞轉錄活化基因-3 (signal transducer and activator of transcription 3, STAT3)，Erk1/2的活化在短時間內(5至10分鐘) 反而減少，STAT3的活化則沒有差異。由此推論，新合成雙酚化合物的保護作用是透過減少ROS的產生，並可以加強NGF對於PC12細胞的神經突生長，但不是透過Erk1/2或STAT3路徑所致。
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描述 碩士
國立政治大學
神經科學研究所
102754005
資料來源 http://thesis.lib.nccu.edu.tw/record/#G0102754005
資料類型 thesis
dc.contributor.advisor 詹銘煥zh_TW
dc.contributor.author (Authors) 林芊瑜zh_TW
dc.creator (作者) 林芊瑜zh_TW
dc.date (日期) 2015en_US
dc.date.accessioned 1-Sep-2015 16:18:16 (UTC+8)-
dc.date.available 1-Sep-2015 16:18:16 (UTC+8)-
dc.date.issued (上傳時間) 1-Sep-2015 16:18:16 (UTC+8)-
dc.identifier (Other Identifiers) G0102754005en_US
dc.identifier.uri (URI) http://nccur.lib.nccu.edu.tw/handle/140.119/78090-
dc.description (描述) 碩士zh_TW
dc.description (描述) 國立政治大學zh_TW
dc.description (描述) 神經科學研究所zh_TW
dc.description (描述) 102754005zh_TW
dc.description.abstract (摘要) 人類隨著年齡增長後中樞神經系統的修補及再生能力逐漸下降，一旦神經系統受到傷害，是很嚴重的問題。因此，引導或促進神經細胞生長甚至再生的方法，中樞神經受損患者將獲得更有效的治療。先前已有文獻指出由植物厚朴萃取的天然化合物─和厚朴酚，具有抗氧化、抗腫瘤、抗發炎、神經保護與滋養的作用。在不同疾病模式的囓齒動物實驗，如帕金森氏症、阿茲海默症、癌症與腦缺血疾病等，和厚朴酚皆具有預防疾病或減緩症狀的效果。本篇研究使用和厚朴酚之衍生物─新合成雙酚化合物(MH102、MH103、MH104、MH106、MH107與MH111)，並探討對於神經細胞的保護與滋養作用。透過腎上腺髓質嗜鉻細胞瘤 PC12 細胞預先處理新合成雙酚化合物，並以過氧化氫(H2O2)使細胞產生氧化壓力，使用活性氧檢測試驗(DCFH-DA assay)偵測細胞內活性氧(reactive oxygen species, ROS)的含量。實驗結果顯示，預先處理較高濃度(3-10μM)的新合成雙酚化合物顯著降低過氧化氫所產生的氧化壓力。另以H2O2誘導PC12細胞死亡，並使用MTT試驗法，觀測新合成雙酚化合物對於細胞存活的影響。結果顯示新合成雙酚化合物顯著減少H2O2造成的細胞死亡。於神經滋養實驗，發現新合成雙酚化合物無法直接誘導PC12細胞的神經突生長。因此，使用神經滋養因子(nerve growth factor, NGF)誘導PC12細胞神經突生長，發現新合成雙酚化合物在低濃度(0.1-0.3μM)顯著加強神經突生長。然而雙酚化合物加強NGF誘導神經突生長之機制，並非透過活化細胞外信號調節激酶 (extracellular-signal-regulated kinases, Erk1/2)與訊息傳遞轉錄活化基因-3 (signal transducer and activator of transcription 3, STAT3)，Erk1/2的活化在短時間內(5至10分鐘) 反而減少，STAT3的活化則沒有差異。由此推論，新合成雙酚化合物的保護作用是透過減少ROS的產生，並可以加強NGF對於PC12細胞的神經突生長，但不是透過Erk1/2或STAT3路徑所致。zh_TW
dc.description.tableofcontents 謝誌 III
中文摘要 V
Abstract VI
目錄 VIII
表次 XI
圖次 XII
縮寫與中英對照表 XIII
第一章、 緒論 1
第一節、 前言 1
第二節、 嗜鉻性細胞瘤細胞 (PC12 cell) 1
一、 PC12細胞簡介 1
二、 PC12細胞之分化 1
第三節、 神經生長因子 (Nerve growth factor, NGF) 3
第四節、 雙酚化合物 (Biphenol compounds) 5
一、 厚朴分化合物 5
二、 和厚朴酚 (Honokiol) 6
三、 和厚朴酚之衍生物 6
第五節、 自由基簡介 (Free radical) 7
一、 活性氧 7
二、 抗氧化系統 7
三、 抗氧化物質 (Antioxidant) 8
第六節、 細胞外信號調節激酶 (Extracellular-signal-regulated kinases, Erk1/2) 9
第七節、 訊息傳遞轉錄活化基因-3 (Signal transducer and activator of transcription 3, STAT3) 10
第八節、 實驗目的與假說 11
第二章、 實驗材料與方法 12
第一節、 化學藥品與儀器 12
一、 化學藥品 12
二、 藥品製備 14
三、 儀器設備 16
第二節、 細胞培養 18
一、 細胞株培養 18
二、 繼代培養與計數 19
三、 細胞冷凍保存 19
四、 細胞解凍 19
第三節、 藥物處理 20
第四節、 活性氧檢測試驗 21
第五節、 細胞存活率試驗法─MTT試驗法 22
第六節、 神經突生長 (Neurite outgrowth assay) 23
第七節、 西方點墨法 23
一、 蛋白質萃取 23
二、 蛋白質濃度測定 23
三、 蛋白質樣品配置 24
四、 鑄膠 24
五、 十二烷基硫酸鈉聚丙烯醯胺凝膠電泳 24
六、 轉漬 25
七、 免疫轉漬 25
第八節、 實驗數據分析 26
第三章、 實驗結果 27
第一節、 過氧化氫引起嗜鉻性細胞瘤細胞PC12之氧化壓力 27
第二節、 雙酚類合成物對於H2O2處理後 ROS 含量之影響 30
第三節、 雙酚類合成物對於H2O2處理後細胞存活率之影響 47
第四節、 雙酚類化合物加強NGF對於PC12細胞神經突生長之影響 54
第五節、 MH101對於NGF處理後Erk1/2磷酸化表現之影響 72
第六節、 MH101對於NGF處理後STAT3磷酸化表現之影響 77
第四章、 討論 80
第五章、 結論 84
參考文獻 85		zh_TW
dc.format.extent 4320511 bytes-
dc.format.mimetype application/pdf-
dc.source.uri (資料來源) http://thesis.lib.nccu.edu.tw/record/#G0102754005en_US
dc.subject (關鍵詞) 雙酚化合物zh_TW
dc.subject (關鍵詞) 腎上腺髓質嗜鉻細胞瘤zh_TW
dc.subject (關鍵詞) 神經保護zh_TW
dc.subject (關鍵詞) 神經突生長zh_TW
dc.subject (關鍵詞) 細胞外信號調節激酶(Erk1/2)zh_TW
dc.subject (關鍵詞) 訊息傳遞轉錄活化基因-3(STAT3)zh_TW
dc.title (題名) 雙酚合成物抑制氧化壓力及加強神經生長因子誘導神經突生長zh_TW
dc.title (題名) The novel biphenol compounds inhibit oxidative stress and enhance nerve growth factor (NGF)-induced neurite outgrowthen_US
dc.type (資料類型) thesisen
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