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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 陳朝峰 | |
dc.contributor.author | Pai-Huei Peng | en |
dc.contributor.author | 彭百慧 | zh_TW |
dc.date.accessioned | 2021-06-14T17:17:09Z | - |
dc.date.available | 2008-08-08 | |
dc.date.copyright | 2008-08-08 | |
dc.date.issued | 2008 | |
dc.date.submitted | 2008-07-25 | |
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/41103 | - |
dc.description.abstract | 神經保護的定義有兩方面的意義:第一,是防止神經細胞死亡的策略;第二,是保護健康但易受傷害的神經細胞。這種保護對神經細胞特別重要,因為一旦神經細胞死亡便無法再生。
視網膜缺血性的傷害是造成視網膜神經節細胞死亡(retinal ganglion cells, RGCs)的重要原因之一,它並且也與一些可能造成失明的眼疾有關,例如缺血性視神經病變、青光眼、及中央視網膜動脈阻塞等。視網膜缺血所造成視網膜神經節細胞死亡的病理機轉主要有幾方面:包括興奮性毒性、發炎、細胞凋亡、及氧化壓力等,尤其是後者,因為再灌流期間大量造成的自由基,造成細胞嚴重的破壞。因此本項實驗的目的是探究神經保護物質的效果;外加的(1)綠茶多酚萃取物epigallocatechin-3-gallate (EGCG),(2)血基質氧化酶-1 Heme oxygenase (HO-1);內生性的, 雌二醇動情激素。並研究其作用機制以期能減少視網膜神經節細胞的傷害,進而維護病患的視覺。 綠茶長久以來被認為具有抗癌、抗炎、及心血管保護作用。最近的文獻顯示它對腦部及視網膜也有神經保護的效果。我們的實驗結果顯示它的確可以減少缺血所造成視網膜神經節細胞死亡,包括細胞凋亡。它的神經保護效果與抑制一氧化氮生成酶的活性有關。 血基質氧化酶-1的作用,會導致prooxidant heme 的分解,產生一氧化碳、鐵離子、及具抗氧化活性的物質-膽綠素。膽綠素會再轉換成也具有抗氧化作用的膽紅素。此過程代表著細胞對於氧化傷害的自我防衛機轉。它的表現會受到許多因子的刺激而大量增加。我們利用藥物的刺激(Cobalt protoporphyrin, CoPP)或是腺病毒基因植入的方式誘發血基質氧化酶-1的大量表現,發現缺血後的視網膜神經節細胞死亡有減少,並且脂質過氧化的程度有降低,抑制細胞凋亡蛋白Bax的表現也減少。 雌二醇動情激素是女性最重要的性激素,它除了傳統的荷爾蒙作用外,近年來更發現其廣泛的存在於中樞神經系統,扮演著調控訊息傳遞等角色。動情激素也具有神經保護及心血管保護的功能。我們利用正常鼠外加雌二醇動情激素,以及去除卵巢的母鼠再予以雌二醇動情激素補充,來評估對視網膜神經節細胞的影響。初步的結果顯示雌二醇動情激素補充,可以挽救去除卵巢後母鼠減少的視網膜神經節細胞。而事前給予雌二醇動情激素可以減少缺血所造成視網膜神經節細胞死亡。 以上的動物實驗結果,提供了視網膜神經細胞保護的初步了解,最終目標是能達到臨床的應用。這有待後續更多的努力。 | zh_TW |
dc.description.provenance | Made available in DSpace on 2021-06-14T17:17:09Z (GMT). No. of bitstreams: 1 ntu-97-D92441002-1.pdf: 1127940 bytes, checksum: 23cb8c191bc22c370575e5d6e347f1ab (MD5) Previous issue date: 2008 | en |
dc.description.tableofcontents | 縮寫表 I
中文摘要 III 英文摘要 V 圖次 VII 表次 IX 第一章、緒論………………………………………………………………………….1 1-1 視網膜…………………………………………………………………………..2 1-2 神經保護……………………………………………………………………... .2 1-2-1 神經保護的定義……………………………………………………………..2 1-2-2 研究眼部視網膜/視神經保護的動物模式…………………………………...3 1-2-3 神經保護的標的………………………………………………………………3 1-2-4 已在使用的降眼壓藥物具有神經保護性質…………………………………5 1-3 視網膜缺血再灌流……………………………………………………………..5 1-4 研究目的………………………………………………………………………..6 第二章、綠茶多酚萃取物Eepigallocatechin-3-gallate (EGCG)經由多種途徑降低視網膜缺血再灌流傷害……………………………………………………………7 2-1 引言………………………………………………………………………………8 2-2 研究材料與方法…………………………………………………………………8 2-2-1 動物處理………………………………………………………………………9 2-2-2 建立視網膜缺血再灌流……………………………………………………..9 2-2-3 視網膜神經節細胞計數……………………………………………………..9 2-2-4 Terminal deoxynucleotidyl-transferase (TdT)-mediated dUTP nick end-labeling (TUNEL method)…………………………………………..10 2-2-5 GFAP、NADPH及一氧化氮生成酶免疫組織化學分析………………….10 2-2-6 以西方點墨法測量GFAP、HO-1、Bcl-2、及Bax的表現……………..11 2-2-7 視網膜脂質過氧化(lipid peroxidation, LPO)的測定………………….12 2-2-8 統計分析……………………………………………………………………12 2-3 結果……………………………………………………………………………13 2-3-1 EGCG減少視網膜缺血再灌流傷害所引起的神經節細胞死亡…………13 2-3-2 EGCG抑制GFAP於視網膜缺血再灌流的活性…………………………13 2-3-3 EGCG減少視網膜神經節細胞的凋亡……………………………………14 2-3-4 EGCG降低神經性一氧化氮生成酶(nNOS)/NADPH的活性……….14 2-3-5 EGCG減少視網膜缺血再灌流後引起的脂質過氧化……………………15 2-3-6 EGCG向上調升血基質氧化酶-1 Heme oxygenase-1(HO-1)的表現.. 15 2-3-7 EGCG不影響Bcl-2及Bax蛋白酶的表現………………………………15 2-4 討論……………………………………………………………………………15 2-5 圖表……………………………………………………………………………20 第三章、誘發血基質氧化酶-1 Heme oxygenase-1(HO-1)可藉由降低氧化壓力及Bax的表現保護視網膜神經節細胞對抗缺血再灌流的傷害………………29 3-1 引言…………………………………………………………………………….30 3-2 研究材料與方法………………………………………………………………31 3-2-1 攜帶HO-1基因腺病毒的合成…………………………………………….31 3-2-2 動物處理……………………………………………………………………32 3-2-3 建立視網膜缺血再灌流……………………………………………………32 3-2-4 以西方點墨法測量HO-1蛋白酶及Bax的表現…………………………32 3-2-5 HO蛋白酶活性的測量…………………………………………………….32 3-2-6 以免疫組織化學法測量HO-1的表現…………………………………….33 3-2-7 視網膜神經節細胞的標記和計數…………………………………………33 3-2-8 視網膜脂質過氧化(lipid peroxidation, LPO)的測定…………………33 3-3 結果……………………………………………………………………………33 3-3-1 Adv-HO-1 基因轉植或給予CoPP向上調升HO-1蛋白酶的表現…….33 3-3-2 給予Adv-HO-1基因或給予CoPP減少視網膜缺血再灌流傷害所引起 神經節細胞死亡…………………………………………………………….34 3-3-3 CoPP減少視網膜缺血再灌流後引起的脂質過氧化……………………35 3-3-4 CoPP減少視網膜缺血再灌流後引起的Bax向上調升…………………35 3-4 討論……………………………………………………………………………35 3-5 圖表……………………………………………………………………………39 第四章、雌性動情素(Estrogen)對視網膜神經節細胞的保護…………………47 4-1 引言……………………………………………………………………………48 4-2 研究材料與方法………………………………………………………………49 4-2-1 動物處理……………………………………………………………………49 4-2-2 雌性動情素濃度的測量……………………………………………………49 4-2-3 視網膜神經節細胞的標記和計數…………………………………………49 4-2-4 GFAP免疫組織化學分析…………………………………………………-49 4-2-5 西方點墨法分析Bcl-2、Bax及BDNF蛋白的表現………………………-49 4-2-6 卵巢切除後視網膜氧化壓力的測量(脂質過氧化、過氧化岐酶、催化酶 穀胱甘酸還原態與氧化態之比值)………………………………………………-50 4-3 結果……………………………………………………………………………50 4-3-1 不同實驗組血漿中雌性動情素的濃度……………………………………50 4-3-2 17β-E2 減少視網膜缺血再灌流傷害所引起的神經節細胞死亡………..51 4-3-3 卵巢切除後大鼠視網膜GFAP的表現增強………………………………51 4-3-4 給予17β-E2不改變視網膜BDNF、Bcl-2及Bax蛋白質的表現……….52 4-3-5 卵巢切除後視網膜氧化壓力增加…………………………………………52 4-4 討論……………………………………………………………………………53 4-5 圖表……………………………………………………………………………55 結論………………………………………………………………………………….65 參考文獻…………………………………………………………………………….66 | |
dc.language.iso | zh-TW | |
dc.title | 大鼠視網膜神經節細胞內生性及外加性的神經保護 | zh_TW |
dc.title | Endogenous and Exogenous Neuroprotection of Retinal Ganglion Cells | en |
dc.type | Thesis | |
dc.date.schoolyear | 96-2 | |
dc.description.degree | 博士 | |
dc.contributor.oralexamcommittee | 趙效明,謝正勇,呂大文,柯美蘭 | |
dc.subject.keyword | 視網膜神經節細胞, | zh_TW |
dc.subject.keyword | Retinal Ganglion Cells, | en |
dc.relation.page | 79 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2008-07-27 | |
dc.contributor.author-college | 醫學院 | zh_TW |
dc.contributor.author-dept | 生理學研究所 | zh_TW |
顯示於系所單位: | 生理學科所 |
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