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研究生: 盧鳳薇
Lu, Feng-Wei
論文名稱: 綠原酸抑制黑色素生成之研究
Studies of Melanogenesis inhibited behavior in B16 Melanoma Cells by Chlorogenic acid
指導教授: 朱一民
Chu, I-Ming
口試委員:
學位類別: 碩士
Master
系所名稱: 工學院 - 化學工程學系
Department of Chemical Engineering
論文出版年: 2009
畢業學年度: 97
語文別: 中文
論文頁數: 56
中文關鍵詞: 綠原酸黑色素酪胺酸脢
外文關鍵詞: chlorogenic acid, melanin, tyrossinase
相關次數: 點閱:3下載:0
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  •   綠原酸(CGA,Chlorogenic acid)是一種廣泛存在於自然界各植物中的一種分子,擁有多種異構物,最常見的為5-咖啡醯奎尼酸(5-O-caffeoylquinic acid),俗名仍同樣通稱為『綠原酸』。除了本身即是一種抗氧化物,目前也有許多文獻指出綠原酸尚具有控制體重、預防癌症、糖尿病或心血管疾病等能力。
      在本研究中,我們的研究是著重在綠原酸是否具有美白潛力,並以老鼠黑色素瘤細胞(B16F0)做為測試平台,發現在不影響細胞生長型態的前提下,讓細胞與含有綠原酸濃度0.6 mM的培養液接觸兩到三天之後,細胞內黑色素含量與酪胺酸□活性會減少為控制組的72%和61%,與此同時,酪胺酸□的基因表現和蛋白質數量也會減少,而所有的抑制效果皆與細胞接觸的綠原酸濃度成正比。此外,我們亦使用雙倒數作圖法(Lineweaver-Burk)觀察綠原酸在酵素反應中的抑制行為,確認其為非競爭型抑制劑。自我們的研究結果中,我們發現綠原酸能夠透過抑制酪胺酸□基因表現,減少酪細胞內的胺酸□蛋白質數量,進而減低最後的黑色素生成量。而我們的研究成果也證明綠原酸具有擔任美白藥物的潛力。


    目錄 摘要 I Abstract Ⅱ 目錄 Ⅲ 第一章 緒論 1   1.1前言 2   1.2皮膚構造 3     1.2.1表皮組織 4     1.2.2真皮組織 6     1.2.3皮下組織 7   1.3黑色素生成路徑 7   1.4酪胺酸脢、TRP-1與TRP-2 11   1.5抑制黑色素的機制與常見美白藥物 12   1.6酵素活性的抑制行為 16   1.7綠原酸簡介 17   1.8研究動機 20 第二章 實驗藥品與研究方法 22   2.1實驗藥品 22   2.2研究方法 24     2.2.1老鼠黑色素瘤細胞培養 24     2.2.2老鼠黑色素瘤細胞內黑色素測試 24     2.2.3黑色素定量曲線 25     2.2.4老鼠黑色素瘤細胞內酪胺酸□活性測試 26     2.2.5 MTT assay細胞存活率測試 27     2.2.6西方點墨法(Western Blot) 28     2.2.7 Lineweaver-Burk Plot 29     2.2.8基因表現分析 30     2.2.9統計分析 31 第三章 實驗結果與討論 33   3.1綠原酸對老鼠黑色素瘤細胞存活率之影響 33   3.2綠原酸對黑色素瘤細胞內黑色素含量之影響 35   3.3綠原酸對於酪胺酸□酵素之影響 38    3.3.1細胞內酪胺酸□酵素之相對活性 38    3.3.2綠原酸對細胞內酪胺酸□基因表現之影響 39    3.3.3綠原酸對細胞內酪胺酸脢數量之影響 43    3.3.4綠原酸對酪胺酸□酵素的直接抑制效果與抑制行為 46 第四章 總結與未來展望 49 第五章 參考文獻 51 附錄 56

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