OXIDATIVE STRESS IN PARKINSON'S DISEASE

帕金森病中的氧化应激

• 批准号: 6594121
• 负责人: JAMES PEPPER BENNETT
• 金额: $ 4.44万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-04-13 至 2005-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6594121
• 关键词: BCL2 gene /protein; Parkinson's disease; apoptosis; brain; calcium flux; cell free system; cellular pathology; cytochrome c; electron transport; enzyme activity; human tissue; hybrid cells; membrane potentials; mitochondria; mitogen activated protein kinase; molecular pathology; mutant; nuclear factor kappa beta; oligonucleotides; oxidative stress; polymerase chain reaction; postmortem; protein protein interaction; transcription factor; transfection /expression vector

Idiopathic Parkinson's Disease (PD) is a major neurodegenerative disease affecting at least 1 million Americans, and the cellular cause of PD is not yet known with certainty. This proposal will explore further the central hypothesis that defects in mitochondrial electron transport chain (ETC) function are a major contributor to premature cell death in PD and will address four Specific Aims 1) define the pathophysiology of mitochondrial transition pore function, and how regulation of membrane potential and intracellular calcium signaling are altered in PD; 2) determine mechanisms of Bcl protein regulation in PD cybrids, and whether transfection with Bcl-overexpression vectors alters mitochondrial function and improves survival; 3) further define the interactions among MAPKinase signaling pathways and NFkappaBeta transcription factor in PD; and 4) characterize mitochondrial transition pore complexes isolated from human postmortem PD brain and compare their function to those isolated from control brain. This project will make use of state-of-the- art intracellular ion imaging technology, RT-PCR techniques, gene transfection strategies, and will develop cell-free systems to examine several inter-related hypotheses. Behind all of these laboratory experiments is a therapeutic imperative, which will be explored in cell and cell-free models. Because new data presented in this application supports the hypothesis of systemically increased oxidative stress in PD patients, exploring these events in an established cell model is even more compelling. This proposal will also compare findings in PD cybrids with those in SY5Y cells exposed to chronic rotenone treatment, a pharmacological cell-based model of complex I loss. Ultimately, the results from this proposal will establish the central importance of genetically acquired mitochondrial ETC dysfunction as an etiologic factor in sporadic PD. Paradigms for evaluating neuroprotective therapies will also be developed to allow targeted approaches to correcting consequences of increased oxidative stress in cells.

特发性帕金森氏病（PD）是一种影响至少100万美国人的主要神经退行性疾病，PD的细胞原因尚不确定。 该提案将进一步探讨一个中心假设，即线粒体电子传输链（ETC）功能的缺陷是PD中过早细胞死亡的主要因素，并将针对四个具体目标1）定义线粒体过渡孔的病理生理学，以及对膜电位和细胞内校立信号的调节如何改变PD中的膜潜在和细胞内钙信号的调节。 2）确定PD Cybrids中BCL蛋白调节的机制，以及用BCL超过表达载体转染是否会改变线粒体功能并提高生存率； 3）进一步定义了PD中Mapkinase信号通路和NFKappabeta转录因子之间的相互作用； 4）表征从人类验尸PD脑分离的线粒体过渡孔复合物，并将其功能与从对照脑分离的功能进行比较。 该项目将利用最先进的细胞内离子成像技术，RT-PCR技术，基因转染策略，并将开发无细胞的系统来检查几种相互关联的假设。 所有这些实验室实验的背后是治疗性命令，将在细胞和无细胞模型中进行探讨。 由于本应用程序中介绍的新数据支持PD患者全身增加的氧化应激的假设，因此在已建立的细胞模型中探索这些事件更具吸引力。该建议还将将PD Cybrid中的发现与SY5Y细胞中的发现结果进行比较，该细胞暴露于慢性葡萄干治疗（一种基于药理学的Conpectial I损失的药理细胞模型）。 最终，该提案的结果将确定遗传获得的线粒体和功能障碍作为零星PD的病因学因素的核心重要性。 还将开发用于评估神经保护疗法的范例，以允许有针对性的方法纠正细胞中氧化应激增加的后果。