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）：NeuroGlobin（NGB）是脊椎动物大脑中最近发现的组织球蛋白。最初的研究表明，NGB对缺氧 - 缺血性侮辱具有保护作用。然而，NGB神经保护基础的机制仍有待定义。 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)下调细胞死亡信号传导和谷氨酸释放减少。在AIM 1中，我们将检查NGB在体外缺氧/二氧化（H/R）后调节线粒体呼吸和神经元死亡信号传导中的作用。我们假设在小鼠脑皮质神经元中H/R后NGB上调，并且过表达NGB可改善线粒体呼吸，并下调神经元死亡信号传导，例如细胞色素C释放和caspase激活。在AIM 2中，我们将记录NGB在体外H/R之后NGB的特定作用。 NGB将在从NNOS敲除小鼠中分离出来的神经元和匹配野生型小鼠的神经元中过表达。将检查并比较RN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AIM 3中，我们将在体内局灶性脑缺血后将NGB过表达的转基因小鼠与野生型小鼠进行比较。将测量神经系统结局，ATP水平，RN和ROS形成，细胞死亡信号传导和谷氨酸释放的改变。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敲除小鼠交叉。这些提出的实验应提供有关NGB如何保护神经元免受缺氧/缺血的新见解，并最终可能导致中风治疗的新型治疗策略。