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：
7159366
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/7159366
关键词：
3-nitrotyrosine Acute Affect Affinity Apoptotic Bioenergetics Biological Assay Brain Brain Hypoxia-Ischemia Brain Injuries Caspase Cell Death Signaling Process Cells Cerebral Ischemia Cerebrum Cessation of life Condition Data Diffusion Edema Extravasation Functional disorder Globin Glucose Glutamates Histology Hypoxia Immunohistochemistry In Situ Nick-End Labeling In Vitro Infarction Ischemia Knockout Mice 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 Nitric Oxide Nitrites Outcome Oxidation-Reduction Oxidative Stress Oxygen Perfusion Process Production Protein Overexpression 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

DESCRIPTION (provided by applicant): Neuroglobin (Ngb) is a recently discovered tissue globin found in the vertebrate brain. Initial study suggests that NGB is protective against hypoxia-ischemic insults. However, the mechanisms underlie Ngb neuroprotection remain to be defined. Our over all hypothesis is that after hypoxia/ischemia, elevated Ngb modulates neuron survival by (a) facilitating oxygen diffusion thus improving mitochondrial respiration and function, (b) scavenges neuronal nitric oxide (NO) thus reducing the formation of reactive nitrogen species (RNS) and reactive oxygen species (ROS), and (c) down-regulating cell death signaling and diminishing glutamate release. In Aim 1, we will examine the role of Ngb in regulating mitochondria respiration and neuronal death signaling after hypoxia/reoxygenation (H/R) in vitro. We hypothesize that Ngb is upregulated after H/R in mouse cerebral cortical neurons and over-expressing Ngb improves mitochondria respiration and down-regulates neuronal death signaling such as cytochrome c release and caspase activation. In Aim 2, we will document the specific role of NO-scavenging by Ngb after H/R in vitro. Ngb will be overexpressed in neurons isolated from nNOS knockout mice and matching wild-type mice. Changes of RNS and ROS formation, mitochondrial respiration, neuronal death signaling and glutamate release will be examined and compared. In Aim 3, we will compare Ngb overexpressing transgenic mice with wild type mice after focal cerebral ischemia in vivo. Alterations of neurological outcomes, ATP levels, RNS and ROS formations, cell death signaling, and glutamate release will be measured. To further dissect the role of Ngb in regulating neuronal NO-mediated brain damage, Ngb transgenic mice will be crossed with nNOS knockout mice. These proposed experiments should provide new insight of how Ngb protect neurons from hypoxia/ischemia and may ultimately lead to novel therapeutic strategies for the treatment of stroke.

描述（由申请人提供）：神经球蛋白（Ngb）是最近在脊椎动物大脑中发现的组织球蛋白。初步研究表明 NGB 对缺氧缺血性损伤具有保护作用。然而，Ngb 神经保护的机制仍有待确定。我们的总体假设是，缺氧/缺血后，Ngb 升高通过以下方式调节神经元存活：(a) 促进氧扩散，从而改善线粒体呼吸和功能；(b) 清除神经元一氧化氮 (NO)，从而减少活性氮 (RNS) 的形成）和活性氧（ROS），以及（c）下调细胞死亡信号并减少谷氨酸释放。在目标 1 中，我们将研究 Ngb 在体外缺氧/复氧 (H/R) 后调节线粒体呼吸和神经元死亡信号传导中的作用。我们假设小鼠大脑皮层神经元中 H/R 后 Ngb 上调，并且过度表达 Ngb 可改善线粒体呼吸并下调神经元死亡信号，例如细胞色素 c 释放和 caspase 激活。在目标 2 中，我们将记录 Ngb 在体外 H/R 后清除 NO 的具体作用。 Ngb 将在从 nNOS 敲除小鼠和匹配的野生型小鼠中分离的神经元中过度表达。将检查和比较 RNS 和 ROS 形成、线粒体呼吸、神经元死亡信号传导和谷氨酸释放的变化。在目标 3 中，我们将在体内局灶性脑缺血后比较 Ngb 过表达转基因小鼠与野生型小鼠。将测量神经系统结果、ATP 水平、RNS 和 ROS 形成、细胞死亡信号传导和谷氨酸释放的变化。为了进一步剖析 Ngb 在调节神经元 NO 介导的脑损伤中的作用，Ngb 转基因小鼠将与 nNOS 敲除小鼠杂交。这些拟议的实验应该为 Ngb 如何保护神经元免受缺氧/缺血提供新的见解，并可能最终带来治疗中风的新治疗策略。