Neuroprotective Mechanisms of Neuroglobin in Stroke

神经球蛋白在中风中的神经保护机制

基本信息

批准号：
7342901
负责人：
XIAOYING WANG
金额：
$ 33.75万
依托单位：
MASSACHUSETTS GENERAL HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2006
资助国家：
美国
起止时间：
2006-01-01 至 2010-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7342901
关键词：
3-nitrotyrosine Acute Affect Affinity Apoptotic Attention Bioenergetics Biological Assay Biological Markers Brain Brain Hypoxia-Ischemia Cell Death Signaling Process Cells Cerebral Ischemia Cerebrum Cessation of life Condition Data Diffusion Edema End Point Extravasation Functional disorder Globin Glucose Glutamates Histology Hypoxia Immunohistochemistry In Situ Nick-End Labeling In Vitro Infarction Ischemia Lactate Dehydrogenase Lead Life Lipid Peroxidation Magnetic Resonance Imaging Malondialdehyde Measurement Measures Mediating Membrane Potentials Mitochondria Modeling Mus Neurologic Neurological outcome Neuronal Hypoxia Neurons Neurotransmitters Nitrates Nitrites Outcome Oxidation-Reduction Oxidative Stress Oxygen Perfusion Process Production Reactive Nitrogen Species Reactive Oxygen Species Recovery Recovery of Function Reporting Research Personnel Respiration Respiratory Chain Reverse Transcriptase Polymerase Chain Reaction Role Rotarod Performance Test Score Signal Transduction Staging Staining method Stains Standards of Weights and Measures Stress Stroke Superoxides Surrogate Markers Testing Time Tissues Transgenic Mice Transgenic Organisms Week Weight Western Blotting Wild Type Mouse apoptosis inducing factor base caspase-3 caspase-9 clinically relevant complex IV cytochrome c cytotoxicity day deprivation excitotoxicity functional outcomes improved in vivo insight mitochondrial membrane morris water maze neuroglobin neuroprotection nitrate novel novel therapeutics programs protective effect research study response

项目摘要

Neuroglobin (Ngb) is a recently discovered tissue globin with a high affinity for oxygen expressed in the vertebrate brain. Initial observations suggest that Ngb is neuroprotective against hypoxic-ischemicinsults. However, the underlying neuroprotective mechanisms remain to be defined. We propose 3 aims to investigate the overall hypothesis that under hypoxia/ischemia conditions, elevated Ngb improves mitochondria respiration, reduces ROS and RNS formation, inhibits cell death signaling, and diminishes glutamate release processes that modulate survival. In Aim 1, we will examine the role of Ngb in regulating mitochondria function and oxidative stress- mediated neuronal death after oxygen-glucose deprivation and hypoxic insults in vitro. Biomarkers of mitochondria integrity, ROS, and NO/ RNS production, cell death signaling, and cytotoxicity will be examined. Responses in Ngb over-expressing neurons will be compared with control (wild-type) mouse cortical neurons, with special attention to baseline levels in the Ngb transgenic neurons. In Aim 2, we will assess neuroprotective effects of Ngb in cerebral ischemia in vivo. Profiles of Ngb expression will be examined after transient (2 hrs) focal cerebral ischemia in mice by western blot, immunohistochemistry and RT-PCR. Mitochondria ATP levels, ROS and RNS production, cell death signaling, glutamate release will be examined. All measurements of baselines and changes after stroke will be compared in Ngb over-expressing transgenic mice versus wild-type littermates. In Aim 3, we will investigate roles of Ngb in neurological outcome and functional recovery after cerebral ischemia in vivo. After acute focal stroke for 1-6 hrs, clinically relevant surrogate markers of energetic stress will be analyzed by MR imaging. At 3 days, morphological outcomes including infarction, edema and BBS leakage will be measured. A standard battery of tests will be used to assess neurological recovery. Baselines and changes of endpoints for Ngb over-expressing transgenic mice will be compared against matching wild-type littermates after permanent and transient 2 hr focal cerebral ischemia. These proposed experiments should provide new insight into how Ngb protects neurons from hypoxia/ischemia and may ultimately lead to novel therapeutic strategies for the acute treatment of stroke.

神经球蛋白（Ngb）是最近发现的一种组织珠蛋白，对氧表达具有高亲和力脊椎动物的大脑。初步观察表明，Ngb 对缺氧缺血性损伤具有神经保护作用。然而，潜在的神经保护机制仍有待确定。我们提出3个目标研究总体假设，即在缺氧/缺血条件下，Ngb 升高可改善线粒体呼吸，减少 ROS 和 RNS 形成，抑制细胞死亡信号传导，并减少调节生存的谷氨酸释放过程。在目标 1 中，我们将研究 Ngb 在调节线粒体功能和氧化应激中的作用 - 体外缺氧和缺氧损伤后介导神经元死亡。生物标志物线粒体完整性、ROS 和 NO/RNS 产生、细胞死亡信号传导和细胞毒性将受到影响。检查了。 Ngb 过度表达神经元的反应将与对照（野生型）小鼠进行比较皮质神经元，特别注意 Ngb 转基因神经元的基线水平。在目标 2 中，我们将评估 Ngb 在体内脑缺血中的神经保护作用。 Ngb简介将在小鼠短暂（2小时）局灶性脑缺血后通过蛋白质印迹检查表达，免疫组织化学和 RT-PCR。线粒体 ATP 水平、ROS 和 RNS 产生、细胞死亡将检查信号传导、谷氨酸释放。所有基线测量和中风后的变化都将在 Ngb 过表达转基因小鼠与野生型同窝小鼠中进行比较。在目标 3 中，我们将研究 Ngb 在脑卒中后神经系统结果和功能恢复中的作用体内缺血。急性局灶性中风 1-6 小时后，能量应激的临床相关替代标志物将通过 MR 成像进行分析。 3 天时，形态学结果包括梗塞、水肿和 BBS 将测量泄漏。将使用一系列标准测试来评估神经功能恢复情况。 Ngb 过表达转基因小鼠的基线和终点变化将与永久性和短暂性 2 小时局灶性脑缺血后与野生型同窝小鼠相匹配。这些拟议的实验应该为 Ngb 如何保护神经元免受缺氧/缺血，可能最终导致中风急性治疗的新治疗策略。