2026-11-092026-11-09https://scholars.csmu.edu.tw/handle/123456789/38780Parkinson’s disease (PD) is one of the neurodegenerative diseases. In addition to motor dysfunction, learning and cognitive deficits are also observed in 20-30% of patients with PD, which is named as dementia in Parkinson’s disease (PDD). Although dopaminergic degeneration is the major causes of PD, neuroinflammation, increase of cytokines, for example, tumor necrosis factor (TNF)-α and interleukin (IL)-2, and glutamatergic hyperactivation are also involved. However, the relationships between neuroinflammation, glutamatergic system, and molecular biological mechanisms are still unclear. Depletion of dopaminergic system by microinjection of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) into the substantia nigra pars compacta (SNc) of rats is a widely used PD animal model. Clinical data shows that D-cycloserine (DCS), a partial agonist of N-methyl-D-aspartate (NMDA) receptor, is able to regulate emotional, learning, and cognitive functions with well tolerance. Recent molecular biological research has reported that signaling pathway of extracellular signal-regulated kinase (ERK)-1/2 is involved in the interaction of NMDA receptors, TNF-α, and IL-2. The aims of this study are elucidating the role of glutamatergic NMDA receptors and neuroinflammation in behavioral deficits seen in PDD and providing evidence for the pathophysiological mechanisms of PDD. Thus, animal model of PD induced by microinjection of MPTP into the SNc of male Wistar rats will be used. Behavior and neuroinflammation will be measured after manipulating the functions of NMDA receptor and neuroimmune by systematic administration of DCS and microinjection of TNF-α and IL-2 into the brain tissue. The following experiments will be performed in this 3-year proposal. The first year: (1) To detect the time course of neuronal degeneration, microglial activation, and changes of TNF-α and IL-2 levels after dopaminergic depletion resulted from microinjection of MPTP into the SNc. (2) To establish behavioral animal model of PDD induced by MPTP treatment: analyzing the spatial working memory, emotional behavior, and object recognition by using the paradigms of T-maze, elevated plus-maze, and object recognition tests, respectively. (3) To elucidate the relationships between brain inflammation and behavioral deficits in animals treated with microinjection of lipopolysaccharide into specific brain areas. The second year: To analyze the effects of DCS on working memory, emotional behavior, recognition, episodic-like memory, and neuroimmune function in MPTP-treated rats. The third year: To explore the effects of local perfusion of TNF-α and IL-2 in the brain tissue on the behavior and activity of ERK-1/2. The following objectives are expected to be achieved in this 3-year project: (1) Clarifying the changes of emotional and cognitive functions in the MPTP-treated animals, which will provide evidence for the validity of using MPTP-treated animal as a PDD model. (2) Providing the data showing the causal relationships between MPTP-induced dysfunctions of NMDA receptors, brain inflammation, cytokine levels, and behavioral deficits. (3) Elucidating the role of ERK signaling pathway in the modulation of neuronal and behavioral functions by cytokines and NMDA receptors, which will provide the possible molecular biological insight of PDD.巴金森氏症（Parkinson’s disease）是常見的神經退化性疾病之ㄧ，其症狀除了運 動功能障礙外，臨床上也觀察到20-30%患者會合併出現學習與認知功能缺陷，稱為「巴 金森氏症失智」（dementia in Parkinson’s disease; PDD）。巴金森氏症的主要病理原因是 多巴胺神經系統退化，但是其他腦區也會出現神經發炎、細胞激素（cytokines），例如 tumor necrosis factor (TNF)-α 及interleukin (IL)-2 含量增加及麩胺酸神經系統功能過度活 化（glutamatergic hyperactivation）之現象，但是目前並不完全清楚麩胺酸神經系統與神 經發炎現象在PDD 之病理生理角色，更遑論細胞分子機轉。研究上普遍使用 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine （ MPTP ） 注射到大鼠的黑質體緻密區 （substantia nigra pars compacta），破壞多巴胺神經系統以建立巴金森氏症之動物模式。 D-cycloserine（DCS）是N-methyl-D-aspartate（NMDA）受體之部分拮抗劑（partial agonist），可以調節與NMDA 受體相關之情緒、學習及認知行為功能，臨床數據顯示 此藥物具有相當高的安全性。近年分子生物學之研究顯示extracellular signal-regulated kinases 1 and 2（ERK1/2）訊息路徑參與調節麩胺酸神經系統NMDA 受體與TNF-α 及 IL-2 間之交互作用。 本研究計畫擬以MPTP 注射到Wistar 大鼠的黑質體緻密區，建立巴金森氏症之動 物模式，並藉由系統性投予DCS 以操弄NMDA 受體之活性及給予大鼠腦內局部注射 TNF-α 及IL-2 以探討麩胺酸神經系統、神經發炎與行為缺陷間之關係，期能提供基礎醫 學之證據以便更清楚瞭解PDD 之病理生理機轉。本項三年期計畫擬進行之實驗如下： 第一年：（1）測量以MPTP 破壞黑質紋狀體多巴胺神經系統後，腦組織內神經 退化、微膠細胞活化及細胞激素TNF-α 及IL-2 變化之時序（time course）；（2）建立 MPTP 巴金森氏症及PDD 之動物行為模式：以T 形迷宮、高腳十字迷宮及物件辨識試 驗分別測量MPTP 處理之動物其空間工作記憶（spatial working memory）、情緒行為及 物件辨識功能；（3）以lipopolysaccharide 注射到特定腦區引起局部發炎及釋放細胞激 素，而後測定動物之上述行為表現，以檢定腦內局部發炎與行為缺陷之關係。第二年： 測量DCS 對MPTP 處理之大鼠的上述行為表現與「類情境記憶」（episodic-like memory） 之影響，並探討DCS 對相關腦區內神經細胞、微膠細胞與細胞激素之影響。第三年： 探討將TNF-α 及IL-2 直接注射到特定腦區對大鼠行為及ERK1/2 訊息路徑之影響。 經由這一系列研究預期可以達到下列目標：（1）提供證據釐清MPTP 巴金森氏症 動物模式是否兼具情緒與認知行為缺陷，這將有助於推論以MPTP 處理之動物做為PDD 動物模式之「效度」（validity）。（2）提供證據顯示MPTP 引起NMDA 受體功能失調、 腦組織發炎、釋出細胞激素與行為缺陷間之「因果關係」（causal relationship）。（3） 瞭解ERK 訊息路徑是否參與細胞激素與NMDA 受體調節神經及行為功能之角色，提供 進一步探討PDD 分子生物學機轉之可能性。巴金森氏症失智症細胞激素麩胺酸神經系統D-cycloserine神經發炎extracellular signal-regulated kinaseParkinson’s diseasedementiacytokineglutamatergic systemD-cycloserineneuroinflammationextracellular signal-regulated kinaseProject