| 姓名 |
蔡昆達(Kun-ta Tsai)
查詢紙本館藏 |
畢業系所 |
物理學系 |
| 論文名稱 |
以光激發黑色素來清除細胞環境中之活性氧之探討
(Verification of melanin function as a light activated ROS scavenger)
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| 相關論文 | |
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| 摘要(中) |
黑色素是一種由許多有機物質構成的一種異質聚合物，其廣泛的分佈在動物體表及體內的許多器官。在過去的一些研究當中，含有黑色素的細胞被認為是保護動物體避免紫外光傷害以及皮膚癌的重要角色。黑色素的保護機制一部分來自於它對可見光波段具有寬廣的吸收帶，在生物體中扮演著光屏蔽的作用，另一方面黑色素也被認為其具有一些更微觀的機制來清除細胞環境中的自由基或是活性氧。在此研究中，我們使用從烏賊萃取出的黑色素顆粒加入至人體的Retinal Pigment Epithelial (RPE) 細胞中，將此做為實驗組，無外加黑色素顆粒者為對照組，在外加過氧化氫來控制活性氧起始濃度的狀況下，將這些細胞培養在一個可提供外部光源照光的自製培養箱內來進行實驗。此研究中使用計數玻片來計算細胞的存活率，以及活性氧指示劑和螢光顯微鏡來做活性氧定量的量測。透過比較細胞存活率以及活性氧含量隨時間的結果顯示，以光來激發黑色素可以清除細胞環境中的活性氧，增加細胞的存活率。
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| 摘要(英) |
Melanin is a heterogeneous biological polymer widespread in our skin and body tissue.Some research reports that melanocytes have a key role in protecting our skin from the
damaging effects of UV radiation and in preventing skin cancer. Furthermore, Melanin is believed that it serves as a photo-protective role in vision by absorbing light and a scavenger of free radicals and reactive oxygen species (ROS). Heterogeneous melanin aggregates from Sepia officinalis, a species of cuttlefish, were added to cultured human retinal pigment epithelial (RPE) cells. Cultures in a home-made incubator were continuous irradiated by a illuminator with controlling the initial concentration of ROS by adding hydrogen peroxide.Cell viability were performed by hemacytometer and the quantity of intracellular ROS were measured by the use of DCFH-DA, a ROS indicator and fluorescence microscope. Time-dependent measurement of cell viability and ROS were performed on parallel cultures consisting of RPE cells which evenly divided into two cultures, one added Sepia melanin and one containing only native melanin. These experiments conclusively demonstrate that melanin serves as a light-activated ROS scavenger to increasing cell viability.
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| 關鍵字(中) |
★ 黑色素
★ 活性氧 |
關鍵字(英) |
★ ROS
★ melanin |
| 論文目次 |
中 文 摘 要........................................................... I
英 文 摘 要.......................................................... II
致 謝............................................................... III
目 錄................................................................ IV
圖 目 錄.............................................................. VI
第一章 緒論........................................................... 1
第二章 材料與實驗方法................................................. 3
2－1 細胞、黑色素與過氧化氫......................................... 4
2－1－1 RPE細胞................................................. 4
2－1－2 黑色素.................................................. 5
2－1－3 過氧化氫................................................ 5
2－2 自製培養箱與照光系統........................................... 6
2－2－1 自製培養箱的設計與 8-well slide ......................... 6
2－2－2 實驗架設................................................ 6
2－3 活性氧的量測................................................... 8
2－3－1 活性氧指示劑－DCFH-DA ................................... 8
2－3－2 螢光顯微鏡、影像擷取與數據處理.......................... 9
2－4 細胞存活率的量測.............................................. 10
第三章 實驗結果與討論................................................ 12
3－1 過氧化氫濃度對細胞存活率的影響................................ 12
3－2 活性氧指示劑對過氧化氫濃度的量測.............................. 13
3－3 黑色素、活性氧與細胞存活率的相關性............................ 14
第四章 結論與後續工作................................................ 18
附錄.................................................................... 19
附錄一： RPE 細胞培養資料........................................... 19
附錄 二： 自製培養箱設計圖 ......................................................................................21
參考文獻.................................................................................................................................24
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| 參考文獻 |
【1】 Wulf Dröge, Free radicals in the physiological control of cell function Physiol Rev 82: 47–95, 2002
【2】 Thomas Herrling, Katinka Jung, The role of melanin as protector against free radicals in skin and its role as free radical indicator in hair Spectrochimica Acta Part A 69 (2008) 1429–1435
【3】 J Geng, W Tang, Photoprotection of bacterial-derived melanin against ultraviolet A–induced cell death and its potential application as an active sunscreen European Academy of Dermatology and Venereology 2008, 22, 852–858
【4】 Brandon-Luke L. Seagle, Elzbieta M. Gasyna et al, Photoprotection of human retinal pigment epithelium cells against blue light-induced apoptosis by melanin free radicals from Sepia officinalis Proceedings of the National Academy of Science, 2006, 103: 16644-16648
【5】 Alice Wang, Anthony R. Marino et al, Photoprotection by porcine eumelanin against singlet oxygen production Photochemistry and Photobiology, 2008, 84: 679–682
【6】 K. C. DUNN, A. E. AOTAKI-KEEN et al, ARPE-19, A human retinal pigment epithelial cell line with differentiated properties Exp. Eye Res. 1996 62, 155-169
【7】 Ulrich Schraermeyer and Klaus Heimann, Current understanding on the role of retinal pigment epithelium and its pigmentation Pigment Cell Res, 1999 12.219-236
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| 指導教授 |
陳賜原(Szu-yuan Chen)
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審核日期 |
2011-1-25 |
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