臺灣博碩士論文加值系統

近視是一種常見的眼科疾病，其中又以新加坡，中國，台灣和日本罹患率較高。除了相關的視力喪失和經濟損失，高度近視會導致退化性疾病，如近視性黃斑部病變、視網膜剝離、後鞏膜葡萄腫和失明。
在眼睛後室的軸向增長是近視的表現型標誌，會使得圖像聚焦在視網膜的前面，導致影像模糊。在臨床治療中，阿托品是一種治療近視有效的藥物，可以停止近視進展；但會造成包括散瞳及睫狀肌麻痺等副作用，而且目前對於阿托品為何可影響近視發展的確切機制仍不清楚。
白藜蘆醇是一種天然的多酚類植物抗毒素，在許多研究中指出其對於許多疾病具有治療效果，例如心血管疾病、癌症、抗老化、抗氧化及抗發炎。
在本實驗室先前的研究中發現，在剝奪型近視老鼠眼睛的 cDNA 表現發現，有一些發炎相關基因表現量上升，例如腫瘤壞死因子 α、介白素 1β、介白素 6 和 cFos，因此促使本論文進一步探討近視眼內的發炎相關蛋白表現。
在剝奪型近視的敘利亞倉鼠眼睛中觀察到，第一型膠原蛋白表現量下降，而腫瘤壞死因子 α 和轉化生長因子 β 表現量上升。當施點了白藜蘆醇後，可以發現上述現象有回復效果。另外，在細胞實驗中，使用脂多醣體體刺激人類視網膜色素上皮細胞，單核球趨化蛋白 1 表現量會上升；而預處理白藜蘆醇後，可以降低脂多醣體體刺激的單核球趨化蛋白 1 表現量上升。我們也觀察白藜蘆醇對於脂多醣體體刺激訊息傳遞路經的影響，脂多醣體會刺激磷酸化的ERK1/2 和 Akt 表現量的增加，而預處理白藜蘆醇後，可以降低磷酸化的 ERK1/2 和 Akt 的表現量。本論文為第一篇指出發炎反應可能是導致近視的原因，而白藜蘆醇可以減少剝奪性近視老鼠的近視度數和發炎反應相關的蛋白質，這顯示了白藜蘆醇，可能可以發展成治療近視的藥物。

Myopia is a common eye disease, and the highest prevalence is observed among Singapore, China, Taiwan and Japan. In addition to the associated substantial visual loss and economic loses, high myopia will lead to degenerative diseases such as myopic macular degeneration, retinal detachment, posterior staphyloma and blindness.
Axial elongation of the vitreal chamber of the eye is the phenotypic hallmark of myopia causing images to focus in front of the retina. In clinical therapy, atropine, is an effective drug that stops myopia progression, but it contains side effects including pupillary dilation and cycloplegia. However, the exact mechanism of the development of myopia is still remains unclear.
Resveratrol is a natural polyphenolic phytoalexin, and it has been implicated as a therapeutic agent in a number of major diseases such as cardiovascular, cancer, anti-aging, anti-oxidant and anti-inflammation. Our previously study showed that some makers of inflammation such as TNFα, IL1β, IL6 and cFOS were increased in form-deprivation myopic eye at cDNA microarray data.
In this study showed that collagen 1 was decreased, and diptor, TNFα and TGFβ were increased at form-deprivation myopia of Syrian hamster animal model, and the expression patterns were reversed after resveratrol treated in immunohistochemistry study. In the cell study, we found the chemokine MCP1 level was increased by LPS stimulated APRPE-19 cell. When cell pre-treated with resveratrol, the LPS-induced MCP1 level was reversed. We also observed that phosphorylation of NFκB, ERK1/2 and Akt were increased by LPS stimulation, when cell pre-treated with resveratrol, the three phosphorylated proteins were reversed. The present study first indicates that inflammatory reaction maybe a source of myopia development. The anti-inflammatory effect of resveratrol maybe becomes a candidate of drug to treat myopia.

致謝
目錄 .............................................................................................................................. i
中文摘要 ........................................................................................................................ iii
英文摘要 ......................................................................................................................... v
壹、 前言 ..................................................................................................................... 1
一、 近視 ..................................................................................................................... 1
二、 近視的病因 ......................................................................................................... 4
I. 近距離用眼 ......................................................................................................... 4
II. 家族基因 ............................................................................................................. 5
三、 近視的致病機轉 ................................................................................................. 6
四、 目前臨床上治療近視的藥物 - 阿托品與其作用機制 ................................... 8
五、 近視與發炎的關係 ........................................................................................... 10
六、 脂多醣體體和發炎反應的機制 ...................................................................... 12
七、 白藜蘆醇 ........................................................................................................... 15
貳、 研究動機 ........................................................................................................... 17
參、 材料與方法 ....................................................................................................... 21
一、 細胞培養 ........................................................................................................... 21
二、 動物實驗 ........................................................................................................... 21
三、 白藜蘆醇配製 ................................................................................................... 22
四、 蛋白質定量 ....................................................................................................... 22
五、 西方墨點法 (Western blot) ............................................................................. 23
六、 免疫組織化學染色法 (immunohistochemistry) ............................................. 27
七、 酵素免疫吸附法 (ELISA)............................................................................... 27
肆、 實驗結果 ........................................................................................................... 29
一、 白藜蘆醇對於使用剝奪性近視方法誘導的敘利亞倉鼠近視度數之影響 .. 29
二、 白藜蘆醇對於近視誘導的發炎相關蛋白及調控近視相關蛋白表現的影響34
三、 白藜蘆醇對於脂多醣體體誘導的單核球趨化激素 1 蛋白表現的影響 .... 40
四、 白藜蘆醇對於脂多醣體體刺激的訊息傳遞路徑之影響 .............................. 42
伍、 討論 ................................................................................................................... 47
陸、 參考文獻 ........................................................................................................... 53

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