臺灣博碩士論文加值系統

視網膜感光細胞(photoreceptors)之退化是由許多因素所造成之病理現象。研究者對於引發感光細胞退化複雜之病理機制並不十分瞭解，例如視網膜色素性病變、老年性黃斑部變性(age-related macular degeneration)、視網膜剝離、光傷害(photic injury)和缺血性傷害等病症。近年來，許多研究學者報告也指出視網膜感光細胞會因為遺傳性基因病變或光傷害而進行自發性凋亡(apoptosis)。基於上述理論基礎，本研究擬評估Minocycline對Lewis albino老鼠視網膜光傷害是否具預防及治療之效果。取6-8週大的Lewis albino老鼠於光循環下飼養2週，且光照實驗前先行暗適應24小時。動物分成光照組和給藥組，將光照組進行光照1, 6, 12, 24小時，光照強度為800-900 lux，而給藥組則施予腹腔注射minocycline (100mg/kg)。老鼠於光照後，置入暗室進行暗恢復(dark recovery) 24小時，待暗恢復後立即犧牲取下視網膜，並利用Western blot及TUNEL染色方法分析。
結果顯示：光照組和給藥組於光照1、6、12、24小時後，以西方點墨法(Western blotting assay)分析Caspase-3表現，結果顯示Caspase-3在此系統被活化。從病理組織學研究發現，感光細胞的變性和視網膜色素上皮層的破壞隨光照時間延長而增加。而由TUNEL-positive細胞核的定量顯示，TUNEL-positive細胞核的數目隨著光照時間的增加而增加。我們的結果證實早期在形態學上觀察到光傷害啟動感光細胞凋亡的現象，我們相信Minocycline 對光損傷所引發之視網膜感光細胞凋亡，僅具部分抑制效果。
關鍵詞：光傷害、細胞凋亡、Caspase-3、TUNEL染色、西方點墨法

Photoreceptor degeneration is a multifactorial pathologic process. The pathogenetic mechanisms of many forms of photoreceptor degeneration are not definitively known, for example, in retinitis pigmentosa, retinal detachment, photic injury, age-related macular degeneration, ischemic injury and other causes. Recent studies demonstrated that retinal photoreceptor cell died by apoptosis in both inherited and photic injury of retinal degeneration. In this experiment we evaluated the therapeutic efficacy of minocyline in photic injury of Lewis albino rats. Six to eight weeks old Lewis albino rats were reared for 2 weeks in cyclic light and dark adaptation for 24 hr before light exposure. The animals were divided into photic-injury and minocycline-treatment groups. The photic-injury groups were exposed to continuous light for 1 to 24 hours in 800-900 lux of light intensity. A dose of 100mg/kg-body weight of minocycline was injected intraperitoneally in minocycline-treatment groups. Light-exposed rats were killed after 24 hr of dark recovery following light exposure. Retinal damage after light exposure was evaluated by Western blot and the terminal transferase-mediated biotin dUTP nick end labeling (TUNEL) technique for identification of nicked/cleaved nuclear DNA. In photic-injury and minocycline-treatment groups, Western blot analysis showed the Caspase-3 active fragment from neural retina after 1,6,12,and 24 hours of light exposure. The degeneration of photoreceptor cells and retina pigment epithelium(RPE) worsened with the increasing duration of light exposure in histopathological study. The numbers of TUNEL positive nuclei were increase with the increasing duration of light exposure. Our findings confirmed earlier morphologic observations that photic exposure triggered apoptosis of photoreceptor cells. We believe that minocycline has partial inhibitory effects to photoreceptor apoptosis after photic injury.
Keywords：photic injury、apoptosis、Caspase-3、TUNEL-stain、Western blot

正文目錄……………………………………………………………………I
圖目錄………………………………………………………………………II
縮寫表………………………………………………………………………III
中文摘要……………………………………………………………………IV
英文摘要……………………………………………………………………VI
第一章 緒 言…………………………………………………………..1
第一節 眼睛構造簡介………………………………………………1
第二節 眼睛血管障壁之結構………………………………………3
第三節 視網膜光傷害………………………………………………7
第四節 影響視網膜光傷害之因素…………………………………9
第五節 細胞凋亡與壞死……………………………………………11
第六節 Minocycline之簡介……………………………………….14
第七節 研究背景及目的……………………………………………17
第二章 實驗材料與方法………………………………………………18
第一節 實驗材料與儀器……………………………………………18
第二節 實驗方法……………………………………………………21
第三章 結果……………………………………………………………33
第四章 討論……………………………………………………………52
第五章 結論……………………………………………………………56
第六章 參考文獻………………………………………………………57
附錄一、 溶液配製

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