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系統識別號 U0026-0812200914361900
論文名稱(中文) 探討褪黑激素對於MPP+所誘發的粒線體斷裂及Drp1表現量之影響
論文名稱(英文) The effect of melatonin on MPP+-induced mitochondrial fission and Drp1 expression
校院名稱 成功大學
系所名稱(中) 生理學研究所
系所名稱(英) Department of Physiology
學年度 96
學期 2
出版年 97
研究生(中文) 潘伊玲
研究生(英文) I-Ling Pan
電子信箱 s3695403@mail.ncku.edu.tw
學號 s3695403
學位類別 碩士
語文別 英文
論文頁數 36頁
口試委員 口試委員-任卓穎
口試委員-蔣輯武
指導教授-莊季瑛
中文關鍵字 褪黑激素  粒線體 
英文關鍵字 melatonin  MPP+  Drp1 
學科別分類
中文摘要 過去的研究中發現,粒線體功能喪失及氧化壓力是導致帕金森氏症的致病機轉,而MPP+會藉由抑制粒線體內膜電子傳遞鏈上complex I的活性,導致氧化壓力的產生進而使得黑質區投射到紋狀體的多巴胺神經元大量流失,因此常被用於誘發帕金森氏症實驗模式的藥物。近年來研究指出,粒線體的型態呈現動態的變化-不斷地融合和分裂,其中位於粒線體上的Mitofusin 1/2和OPA1主要是調控粒線體之間的融合,而分布於細胞質中的Drp1移至粒線體上和hFis1進行交互作用則是調控粒線體分裂的主要機制。研究結果指出,阻斷Drp1的活化會促進粒線體呈現長條狀並能對抗由活性氧物質所誘發的細胞毒性。此外,我們實驗室過去的研究指出褪黑激素可以藉由其抗氧化的功能有效保護MPP+所誘發的多巴胺神經細胞損傷,然而粒線體型態改變是否參與在褪黑激素的保護機制之中至今仍無人探討。因此我們將探討Drp1是否參與在MPP+誘發的神經毒性及褪黑激素的保護作用之中。首先,我們發現MPP+誘發氧化壓力上升、粒線體斷裂,和粒線體上Drp1蛋白質表現量上升,最後導致神經細胞死亡。為了更進一步探討Drp1所扮演的角色,我們在神經細胞中大量表現Drp1,發現細胞的粒線體會呈現分離斷裂的型態,而在大量表現沒有活性的Drp1K38A時,則可避免MPP+所造成的粒線體斷裂現並降低細胞死亡。另外,我們也發現同時給予褪黑激素可以降低MPP+誘發的氧化壓力、減少粒線體斷裂和粒線體上Drp1蛋白質表現量上升等現象,進而保護神經細胞免於MPP+的毒殺作用。這些結果顯示:在MPP+的處理下,細胞會產生氧化壓力,並透過Drp1造成粒線體斷裂、分離和分解,進而導致神經細胞死亡,而褪黑激素可藉由其抗氧化能力降低MPP+所誘發的氧化壓力而減少粒線體斷裂來達到保護神經細胞的效果。
英文摘要 Mitochondrial dysfunction and oxidative stress are involved in pathogenesis of Parkinson’s disease (PD). 1-methyl-4-phenylpyridinium (MPP+), an inhibitor of mitochondrial complex I, induces oxidative insults on nigrostriatal dopaminergic neurodegeneration and parkinsonism in primates. Mitochondrial morphology is dynamic change and precisely regulated by the mitochondrial fission and fusion machinery. Human Fis1 (hFis1) and the translocation of dynamin-related protein 1 (Drp1) regulate mitochondrial fission, whereas the mitochondrial fusion is regulated by mitofusin 1/2 (Mfn1/2) and optic atrophy 1 (OPA1). The role of Drp1-regulated mitochondrial fission was demonstrated by the results showing that cells overexpressing dominant-negative Drp1K38A were resistant to reactive oxygen species-induced toxicity. Furthermore, our previous studies showed that melatonin functioned as an antioxidant to prevent MPP+-induced neuronal death. However, the effect of melatonin on mitochondrial morphology remains unknown. We therefore investigate whether Drp1 is involved in MPP+-induced mitochondrial fission and the protective effect of melatonin. We found that MPP+-induced oxidative stress, mitochondrial fission, mitochondrial Drp1 upregulation and consequently cell death. Furthermore, we currently observed that overexpressing EGFP-Drp1 in neurons promoted mitochondrial fission, however, overexpression of EGFP-Drp1K38A in neurons blocked MPP+-induced mitochondrial fission and cell death. Moreover, melatonin co-treatment significantly prevented MPP+-induced oxidative insults, mitochondrial fission, mitochondrial Drp1 upregulation and cell death. The results suggest that MPP+ induced oxidative stress, Drp1-mediated mitochondrial fission and finally cell death. However, co-treatment with melatonin and MPP+ protected neurons from MPP+ toxicity through reducing oxidative stress and mitochondrial fission.
論文目次 Abstract………………….……………………………………….…3
Chinese………………….………………………………......3~4
English………………………….……………………………5~6
Table of contents……………………………………………………..7
Acknowledgement………………………………………………………8
Introduction……………………………………………………9~14
Materials and Methods…………….……………………………15~20
Results………………………………………………………….21~26
Discussion………………….……………………………………27~30
References………………….……………………………………31~36
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