ROLE OF PROHIBITIN IN ISCHEMIC BRAIN INJURY

抑制素在缺血性脑损伤中的作用

• 批准号: 7768118
• 负责人: Ping Zhou
• 金额: $ 36.97万
• 依托单位: WEILL MEDICAL COLL OF CORNELL UNIV
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-15 至 2014-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7768118
• 关键词: ATP Synthesis Pathway; Alteplase; Apoptosis; Bioenergetics; Brain; Brain Hypoxia-Ischemia; Brain Injuries; Cerebral Ischemia; Cerebrum; Cessation of life; Clinical; Complex; Cytoprotective Agent; Defense Mechanisms; Down-Regulation; Electrons; Functional disorder; Gene Transfer; Glucose; Health; Hippocampus (Brain); In Vitro; Injury; Ischemia; Ischemic Brain Injury; Ischemic Preconditioning; Ischemic Stroke; Membrane Proteins; Methods; Mitochondria; Mitochondrial Membrane Protein; Modeling; Mus; N-Methylaspartate; Neuronal Injury; Neurons; Organ; Organelles; Oxidative Stress; Oxygen; Patients; Pattern; Phase; Predisposition; Process; Production; Prosencephalon; Proteins; Proteomics; Reactive Oxygen Species; Recombinant adeno-associated virus (rAAV); Recovery; Resistance; Respiratory Chain; Role; Small Interfering RNA; Stimulus; Stroke; Testing; Therapeutic; Time; Up-Regulation; Viral Genes; cell type; deprivation; effective therapy; improved; improved functioning; in vivo; in vivo Model; inhibitor/antagonist; neuronal survival; neuroprotection; new therapeutic target; novel; novel therapeutic intervention; novel therapeutics; overexpression; preconditioning; prohibitin; protective effect; public health relevance; research study; treatment strategy

DESCRIPTION (provided by applicant): Prohibitin (PHB) is a mitochondrial inner membrane protein that may preserve cellular integrity by stabilizing the function of complex I, the electrons entry point into the respiratory chain, and reducing production of mitochondrial reactive oxygen species. In a proteomic study seeking to identify potential neuroprotective proteins expressed in murine models of ischemic tolerance, we found that PHB expression is increased in neuronal mitochondria. These observations raise the possibility that PHB upregulation in preconditioning models promotes neuronal survival, while its downregulation in ischemia facilitates neuronal death. Thus, the long-term objectives of this application are to elucidate the roles of PHB in the brain damage produced by cerebral ischemia and to assess its neuroprotective potential. In particular, we will test the central hypothesis that PHB, by influencing the mitochondrial resistance to injury, is a key determinant of neuronal fate in the ischemic brain. The proposed experiments will use in vitro (oxygen-glucose deprivation), and in vivo (transient forebrain ischemia) models of cerebral ischemic injury. Viral gene transfer and small interfering RNA (siRNA) will be used to increase or decrease PHB expression in neuronal cultures or in the mouse hippocampus. Mitochondrial function will be assessed in neuronal cultures or in isolated mitochondria to explore the mechanisms of the effect of PHB. The following hypotheses will be tested: (a) Hypoxia-ischemia downregulates PHB, a reduction that decreases endogenous defense mechanisms and may increase the susceptibility of the brain to injury; (b) Expression of PHB in neuronal cultures is neuroprotective, while its downregulation increases vulnerability to injury; (c) Expression of PHB in the mouse hippocampus protects vulnerable neurons from the damage produced by transient forebrain ischemia; (d) The mechanisms of the neuroprotective effect of PHB involve complex I stabilization and reduced production of mitochondrial reactive oxygen species. PUBLIC HEALTH RELEVANCE: The proposed studies will investigate a novel aspect of the pathobiology of PHB, related to its role in the death and survival of ischemic neurons. The findings will advance our understanding of the fundamental processes regulating ischemic neuronal death, and have the potential of identifying new treatment strategies for ischemic stroke.

描述(申请人提供)：Prohibitin(PHB)是一种线粒体内膜蛋白，可通过稳定电子进入呼吸链的复合体I的功能并减少线粒体活性氧的产生来维持细胞的完整性。在一项旨在确定在缺血耐受小鼠模型中表达的潜在神经保护蛋白的蛋白质组学研究中，我们发现PHB在神经元线粒体中的表达增加。这些观察结果提出了一种可能性，即在预适应模型中PHB的上调促进了神经元的存活，而在缺血时PHB的下调促进了神经元的死亡。因此，这一应用的长期目标是阐明PHB在脑缺血造成的脑损伤中的作用，并评估其神经保护潜力。特别是，我们将检验中心假设，即PHB通过影响线粒体对损伤的抵抗力，是缺血脑中神经元命运的关键决定因素。拟议的实验将使用体外(缺氧-葡萄糖剥夺)和体内(短暂性前脑缺血)脑缺血损伤模型。病毒基因转移和小干扰RNA(SiRNA)将用于在神经元培养或小鼠海马体中增加或减少PHB的表达。线粒体功能将在神经元培养或分离的线粒体中进行评估，以探索PHB的作用机制。将检验下列假说：(A)缺氧-缺血下调PHB，这种减少减少了内源性防御机制，并可能增加大脑对损伤的敏感性；(B)PHB在神经元培养中的表达具有神经保护作用，而其下调则增加了对损伤的易感性；(C)在小鼠海马区的表达可保护脆弱的神经元，使其免受短暂性前脑缺血造成的损伤；(D)PHB的神经保护作用的机制涉及复杂I稳定和线粒体活性氧物种的产生减少。 公共卫生相关性：拟议的研究将调查PHB病理生物学的一个新方面，与其在缺血神经元死亡和存活中的作用有关。这些发现将促进我们对调节缺血性神经元死亡的基本过程的理解，并有可能确定缺血性中风的新治疗策略。