黑色素的奈米化大幅提升其用在保護細胞對抗活性氧壓力的效用

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姓名

陳思旻(Sih-min Chen) 查詢紙本館藏

畢業系所

生物物理研究所

論文名稱

黑色素的奈米化大幅提升其用在保護細胞對抗活性氧壓力的效用
(Dramatic Improvement of the Efficacy of Melanin on Protecting Cells against ROS Stress by Its Nanonization)

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摘要(中)

目的：黑色素是已知具有清除自由基及活性氧的能力。本論文提出利用奈米化黑色素作為藥物來對抗內源性氧化壓力或外源性急性氧化壓力，例如：急性輻射綜合症。

材料與方法：發展一個簡單的技術，利用機械式攪拌來進行黑色素奈米化，使黑色素具有奈米尺度大小與親水性。存活率測試使用視網膜色素上皮細胞，將其暴露在外源性H2O2壓力下，並以不同條件的黑色素及照光來處理。

結果與討論：機械式攪拌可以有效奈米化黑色素。在視網膜色素上皮細胞暴露在外源性活性氧壓力下，奈米化黑色素相較於未處理的黑色素明顯有效提高存活率。這個結果在不照光時有更明顯的提升，有希望應用於靜脈給藥的體內試驗。

摘要(英)

Aim: It is known that melanin has the ability of scavenging free radicals and reactive oxygen species (ROS). The authors propose to utilize nanonized melanin as medication against endogenous chronic oxidative stress or exogenous acute oxidative stress caused by, e.g., acute radiation exposure.

Materials & Methods: A simple technique based on mechanical stir was developed for nanonization of melanin to produce nanometer-sized and water-dispersible melanin. The viabilities of cultured retinal pigment epithelium (RPE) cells exposed to exogenous H2O2 stress and treated with various conditions of melanin and irradiation were compared.

Results & Conclusion: Melanin can be nanonized very effectively with simple mechanical stir. Nanonized melanin exhibited a much stronger effect than unprocessed melanin on raising the viability of cultured RPE cells under exogenous acute ROS stress. The effect was even more prominent without simultaneous light irradiation, promising for effective in vivo application to the whole body via intravenous administration.

關鍵字(中)

★ 黑色素
★ 奈米薄片
★ 水分散性
★ 活性氧
★ 自由基
★ 氧化壓力
★ 急性輻射綜合症

關鍵字(英)

★ melanin
★ nano-flake
★ water-dispersible
★ reactive oxygen species
★ free radical
★ oxidative stress
★ acute radiation syndrome

論文目次

中文摘要................................ i
英文摘要................................ ii
謝誌....................................... iii
目錄....................................... iv
圖目錄.................................... vi

一、緒論.................. 1
1.1 黑色素. . . . . . . . . . . . . . . . . . . . . . . . . 1
1.2 過氧化氫. . . . . . . . . . . . . . . . . . . . . . . 2
1.3 研究成果. . . . . . . . . . . . . . . . . . . . . . . 2
1.3.1 發展簡易技術使黑色素奈米化. . . . . . . . . . . . . 2
1.3.2 證實奈米化黑色素有能力清除活性氧. . . . . . . . . 2

二、材料與方法....... 3
2.1 黑色素奈米化. . . . . . . . . . . . . . . . . . . . . 3
2.2 原子力顯微鏡. . . . . . . . . . . . . . . . . . . . . 3
2.3 電子順磁共振. . . . . . . . . . . . . . . . . . . . . 4
2.4 細胞培養. . . . . . . . . . . . . . . . . . . . . . . 4
2.5 存活率量測. . . . . . . . . . . . . . . . . . . . . . 5
2.6 H2O2誘導細胞凋亡. . . . . . . . . . . . . . . . . . 5
2.7 黑色素保護細胞對抗H2O2分析試驗. . . . . . . . . 6

三、結果與討論....... 7
3.1 奈米化黑色素診斷. . . . . . . . . . . . . . . . . . 7
3.1.1 製程與沉降時間的吸光度變化. . . . . . . . . . . . . 7
3.1.2 合成黑色素AFM影像. . . . . . . . . . . . . . . . . 8
3.1.3 奈米化合成黑色素直徑及高度分析. . . . . . . . . . 9
3.1.4 吸收光譜及EPR頻譜量測. . . . . . . . . . . . . . . 11
3.2 體外細胞試驗. . . . . . . . . . . . . . . . . . . . . 12
3.2.1 H2O2濃度對RPE細胞存活率. . . . . . . . . . . . . 12
3.2.2 RPE細胞在不同條件下存活率試驗. . . . . . . . . . 13

四、結論與未來展望........15
4.1 結論. . . . . . . . . . . . . . . . . . . . . . . . . 15
4.2 未來展望. . . . . . . . . . . . . . . . . . . . . . . 15
文獻.......................................16
附錄一....................................19

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指導教授

陳賜原(Szu-yuan Chen)

審核日期

2014-8-28

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