臺灣博碩士論文加值系統

隨著社會高齡化，與年齡增長有關的神經退化性疾病患者也逐年增加。其中巴金森氏症 (Parkinson's disease) 的盛行率僅次於阿茲海默氏症 (Alzheimer's disease)，巴金森氏症的主要病理特徵是中腦黑核 (substantia nigra) 之多巴胺神經元不明原因退化死亡，導致多巴胺神經元之神經軸末梢所在處紋狀體 (striatum) 之多巴胺分泌不足，進而引發患者震顫 (tremor)、動作遲緩 (bradykinesia) 和步態異常 (freezing of gait) 等症狀。巴金森氏症發病原因不明，但已知細胞自噬 (autophagy) 異常會引發神經退化性疾病。細胞自噬為細胞內重要的「分解、回收、再利用」系統，主要是將失能蛋白、胞器、受損粒線體運送至溶體 (lysosome) 分解。而活化細胞自噬以清除神經毒性蛋白質糾結 (protein aggregations) 及損傷粒線體俱有神經保護及治療巴金森氏症潛力。目前巴金森氏症治療僅能改善症狀及延緩病程發展，仍無法治癒。針灸為傳統中醫療法，被證實可治療多種疾病，而電針則可將針灸條件標準化。因此本研究評估電針在巴金森氏症小鼠之療效，並且分析電針對不同腦區細胞自噬影響，藉以尋找電針治療巴金森氏症的機轉及對神經退化型疾病之可能應用。本研究證實電針陽陵泉及太衝兩穴位，有效治療巴金森氏症小鼠的動作障礙症狀並抑制多巴胺神經退化死亡，並且可以專一性活化黑核、紋狀體、海馬迴和大腦皮質中神經元細胞自噬體生合成 (autophagosome biogenesis)、受質分解、及粒線體自噬 (mitophagy)。因上述腦區與多種神經退化性疾病的發生有關，顯示運用電針治療巴金森氏症、阿茲海默氏症、亨廷頓舞蹈症 (Huntington's disease) 等俱有臨床應用潛力。

With the advent of an aging society, the number of patients with age-related neurodegenerative diseases is also increasing yearly. The prevalence of Parkinson's disease is second only to Alzheimer's disease. The main pathological manifestations of Parkinson's disease are the gradual degeneration and death of dopaminergic neurons in the substantia nigra of the brain, which result in insufficient dopamine hormones in the striatum and cause symptoms such as tremor, bradykinesia, and freezing of gait. The predisposing factors of Parkinson's disease are unknown, but it is known that the defect of autophagy is one of the important factors causing neurodegenerative diseases. Autophagy is an essential system of decomposition and recycling in cells, which mainly transport incapacitating proteins, organelles, and damaged mitochondria to the lysosomes for breakdown. Enhanced autophagy to clear protein aggregations of neurotoxic proteins and damaged mitochondria has neuroprotective effects. The current treatments only can improve the symptoms but cannot delay the progression of the disease. Acupuncture is a traditional Chinese medicine therapy that has long been proven effective in treating various diseases, and electroacupuncture stimulation can standardize acupuncture conditions. Therefore, we designed a study to assess the efficacy of electroacupuncture in Parkinson's disease model mice and to analyze the degree of autophagy activation in different brain regions after electroacupuncture. We expect to determine the mechanism of electroacupuncture in treating of Parkinson's disease and its applications in neurodegenerative diseases. The results of our study demonstrate that electroacupuncture stimulation on Yanglingquan (GB34) and Taichong (LR3) acupoints can improve the symptoms of dyskinesia and the degeneration of dopaminergic neurons in Parkinson's disease model mice. Moreover, electroacupuncture can specifically activate autophagosome biogenesis, substrate degradation, and mitophagy in substantia nigra, striatum, hippocampus, and cerebral cortex, which are among the main affected brain areas of many patients with neurodegenerative diseases. Therefore, electroacupuncture has potential clinical application in the treatment of Parkinson's disease, Alzheimer's disease, and Huntington's disease.

中文摘要 I
Abstract II
目次 IV
圖表目次 VI
論文正文 1
第一章 前言 1
第一節 研究背景 1
第二節 研究目的 8
第二章 研究材料與方法 9
第一節 巴金森氏症小鼠動物模式 9
第二節 電針 10
第三節 小鼠動作障礙分析-圓筒試驗與加速跑步機試驗 12
第四節 西方墨點 14
第五節 免疫螢光染色與共軛焦顯微鏡分析 16
第六節 統計分析 18
第三章 研究設計與步驟 19
第一節 評估電針治療對於巴金森氏症小鼠之動作障礙及腦部紋狀體細胞自噬影響 19
第二節 評估氣體麻醉對小鼠大腦皮質之細胞自噬影響 20
第三節 評估電針對小鼠腦部黑核、紋狀體、海馬迴、大腦皮質中細胞自噬影響 21
第四章 研究結果 22
第一節 電針治療對巴金森氏症小鼠的動作障礙症狀及紋狀體細胞自噬的影響 22
第二節 氣體麻醉對小鼠大腦皮質之細胞自噬影響 27
第三節 電針對小鼠黑核細胞自噬影響 30
第四節 電針對小鼠紋狀體細胞自噬及粒線體自噬影響 35
第五節 電針對小鼠海馬迴細胞自噬及粒線體自噬影響 41
第六節 電針對小鼠大腦皮質細胞自噬及粒線體自噬影響 47
第五章 討論 53
第一節 研究結果討論 53
第二節 研究限制 59
第六章 結論與建議 60
第一節 結論 60
第二節 未來可能之研究方向 61
參考文獻 62
附錄 70
圖表目次
圖一、電針儀與電針 11
圖二、圓筒試驗和加速跑步機試驗 13
圖三、電針治療巴金森氏症小鼠的動作障礙並活化紋狀體細胞自噬 24
圖四、異氟醚 (isoflurane) 麻醉不影響小鼠大腦皮質細胞自噬 28
圖五、電針活化小鼠中腦黑核多巴胺神經元與其他神經元細胞自噬 31
圖六、電針活化小鼠腦部紋狀體神經元的細胞自噬體生合成、細胞自噬受質水解和粒線體自噬 36
圖七、電針活化小鼠腦部海馬迴神經元的細胞自噬體生合成、細胞自噬受質水解和粒線體自噬 42
圖八、電針活化小鼠大腦皮質區域神經元的細胞自噬體生合成、細胞自噬受質水解和粒線體自噬 48
附圖一、巴金森氏症治療藥物之作用位點 70
附圖二、左旋多巴胺藥效減退現象 71
附圖三、大腦深層電刺激手術 72
附圖四、經顱磁振導航聚焦超音波手術 73
附圖五、細胞自噬分解過程 74
附圖六、吞噬泡形成與細胞自噬體生合成 75
附圖七、粒線體自噬 77
附圖八、細胞自噬與神經退化性疾病的相關性 78
附圖九、動物實驗審查證明 79

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