ROLE OF PROHIBITIN IN ISCHEMIC BRAIN INJURY

抑制素在缺血性脑损伤中的作用

• 批准号: 8130923
• 负责人: Ping Zhou
• 金额: $ 36.23万
• 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/8130923
• 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; spatiotemporal; 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（由申请人提供）：抑制素（PHB）是一种线粒体内膜蛋白，它可以通过稳定复合物I（呼吸链的电子入口点）的功能并减少线粒体活性氧的产生来保持细胞完整性。在一项旨在鉴定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 表达在神经元线粒体中增加。这些观察结果表明，预处理模型中 PHB 上调可促进神经元存活，而缺血中 PHB 下调则促进神经元死亡。因此，本申请的长期目标是阐明 PHB 在脑缺血引起的脑损伤中的作用并评估其神经保护潜力。特别是，我们将检验以下中心假设：PHB 通过影响线粒体对损伤的抵抗力，是缺血性大脑中神经元命运的关键决定因素。拟议的实验将使用脑缺血损伤的体外（缺氧-葡萄糖剥夺）和体内（短暂前脑缺血）模型。病毒基因转移和小干扰 RNA (siRNA) 将用于增加或减少神经元培养物或小鼠海马中的 PHB 表达。将在神经元培养物或分离的线粒体中评估线粒体功能，以探索 PHB 的作用机制。将测试以下假设： (a) 缺氧缺血会下调 PHB，这种减少会降低内源性防御机制，并可能增加大脑对损伤的敏感性； (b) PHB 在神经元培养物中的表达具有神经保护作用，而其下调会增加受伤的可能性； (c) PHB 在小鼠海马中的表达可保护脆弱的神经元免受短暂前脑缺血造成的损害； (d) PHB 的神经保护作用机制涉及复合物 I 稳定和减少线粒体活性氧的产生。 公共卫生相关性：拟议的研究将调查 PHB 病理学的一个新方面，与其在缺血性神经元死亡和存活中的作用有关。这些发现将增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。