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研究生:顏瑄
研究生(外文):YEN, HSUAN
論文名稱:茶鹼促進黑色素合成作用機制的研究
論文名稱(外文):Study of the action mechanism of theophylline on enhancement of melanin synthesis
指導教授:張聰民張聰民引用關係
指導教授(外文):CHANG, TSONG-MIN
口試委員:柯俊良黃蕙君
口試委員(外文):KO, JIUNN- LIANGHUANG, HUEY-CHUN
口試日期:2017-05-12
學位類別:碩士
校院名稱:弘光科技大學
系所名稱:化妝品應用系暨化妝品科技研究所
學門:民生學門
學類:美容學類
論文種類:學術論文
論文出版年:2017
畢業學年度:105
語文別:中文
論文頁數:106
中文關鍵詞:茶鹼酪胺酸酶黑色素抗氧化
外文關鍵詞:TheophyllineTyrosinaseMelaninanti-oxidant
相關次數:
  • 被引用被引用:2
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茶鹼(Theophylline)是一種磷酸二酯酶抑制劑,廣泛用於呼吸系統疾病的治療,具有與咖啡因類似的結構和藥理學特性。曾有研究指出茶鹼可以促進黑色素細胞的生長與黑色素含量,但是作用機制不明。本研究探討茶鹼調控黑色素產生機制,及其抗氧化能力。本研究進行B16F10與B16F1小鼠黑色素瘤細胞中的酪胺酸酶活性和黑色素含量測定,使用西方墨點法測定黑色素生成的相關蛋白質的表現變化,與訊息調控之機制,同時也檢測茶鹼清除自由基能力及抗氧化活性。結果顯示,濃度範圍在100-500 μM的茶鹼,可以有效的增加B16F10細胞內酪胺酸酶活性及黑色素含量。由西方墨點法和訊息調節劑之試驗發現,茶鹼能透過JNK及Wnt/β-catenin訊息傳遞路徑影響蛋白質表現量,刺激黑色素生成。本論文證實茶鹼具有促黑的能力。
Theophylline, a kind of phosphodiesterase inhibitor, is widely used in the treatment of respiratory diseases, has the similar pharmacological properties to those of caffeine. Several studies noted that theophylline could promote the growth of melanocytes and melanin content, whereas the action mechanism is yet be delineated. The aim of this study is to investigate the effects of theophylline on melanoma proliferation and its possible mechanisms. We also evaluate the antioxidation capacity of theophylline. In the present study, the tyrosinase activity and melanin content in B16F10 and B16F1 melanoma cells were measured, the changes of the expression melanogenesis-related proteins in B16F10 melanoma cells were determined by the Western blot. The cell melanin contents were also determined under different kinds of protein kinase inhibitors in cells. We also examined the free radical scaveng and antioxidant activities of theophylline. The results revealed that theophylline(100-500 μM) effectively increased the intracellular tyrosinase activity and the contents of melanin in B16F10 cells. From the results of Western blotting experiments and protein kinase regulators assays, theophylline can influence the amount of melanogenesis-related proteins’ expression and stimulate melanin production through the JNK and Wnt / β-catenin signaling pathways. Therefore, theophylline could act as a new tanning ingredient.
致謝
中文摘要
英文摘要
目錄
圖目錄
壹、緒論
一、茶鹼
二、皮膚與黑色素
三、光保護與光損害
四、黑色素生成途徑
五、黑色素細胞調控的訊息傳遞路徑
 (一)PKA訊息路徑
 (二)MAPK訊息路徑
 (三)Wnt訊息路徑
六、自由基與抗氧化
 (一)自由基與ROS
 (二)抗氧化劑

貳、研究動機與實驗架構
一、研究動機
二、實驗架構
参、材料與方法
一、實驗材料
 (一)藥品來源
 (二)儀器設備
二、實驗方法
 (一)樣品製備
 (二)細胞培養
 (三)細胞存活率分析
 (四)美白能力分析
 (五)抗氧化能力分析
 (六)統計分析
肆、實驗結果
一、美白能力測定
 (一)茶鹼對B16F10與B16F1細胞存活率之影響
 (二)茶鹼對蕈菇酪胺酸酶活性之影響
 (三)茶鹼對B16F10與B16F1細胞黑色素含量之影響
 (四)茶鹼對B16F10與B16F1細胞酪胺酸酶活性之影響
 (五)茶鹼對B16F10細胞內蛋白質表現量之影響
 (六)訊息調節劑對茶鹼影響B16F10細胞黑色素含量之改變
二、抗氧化能力測定
 (一)茶鹼之DPPH自由基清除能力測定
 (二)茶鹼之ABTS‧+自由基清除能力測定
 (三)茶鹼之總酚含量測定
 (四)茶鹼之螯合金屬離子能力測定
 (五)茶鹼之還原力測定
伍、討論
陸、結論
柒、參考文獻
捌、結果附圖

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