Age-related alterations to NADPH oxidase and glial cell activation after spinal cord injury

脊髓损伤后 NADPH 氧化酶和神经胶质细胞活化的年龄相关变化

基本信息

批准号：
9051752
负责人：
Ramona E. von Leden
金额：
$ 2.71万
依托单位：
HENRY M. JACKSON FDN FOR THE ADV MIL/MED
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-12-01 至 2017-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9051752
关键词：
Acute Adult Affect Age Age-Months Aging Apoptotic Astrocytes Biochemical Biochemistry Brain Cell Fractionation Cell Survival Cells Chronic Cicatrix Cods Contusions Data Detection Diagnosis Elderly Enzymes Evaluation Family Future Gene Expression Human Immune Immunohistochemistry Inflammation Inflammatory Inflammatory Response Injury Label Lesion Life Macrophage Activation Measures Microglia Modeling Morphology Motor Mus NADP NADPH Oxidase Neuraxis Neuroglia Neurons Outcome Oxidases Oxidative Stress Phenotype Play Population Production Rattus Reactive Oxygen Species Recovery Recovery of Function Research Personnel Rest Rodent Role Severities Site Source Spinal Spinal Cord Spinal cord injury Stimulus Techniques Testing Tissues Trauma Traumatic Brain Injury Up-Regulation Work age effect age related aged aging population base cell type comorbidity cytokine disability functional outcomes improved inhibitor/antagonist insight middle age neurotoxic public health relevance repaired research study response response to injury stressor therapeutic target young adult

项目摘要

DESCRIPTION (provided by applicant): Over 250,000 people in the US are currently living with disabilities related to spinal cord injury (SCI). The number of SCI's among the aging population has been steadily increasing since the 1980's, and is associated with high rates of co-morbidities. Oxidative stress and reactive oxygen species (ROS) are increased in aging tissue and have been causally implicated in tissue damage and chronic inflammation in the central nervous system (CNS). These stressors influence the activity of glial cells, contributing to the delayed recovery to SCI seen in the aging population. No work to date has examined age-related alterations in glial cell response to SCI. The NADPH oxidase (NOX) family of enzymes is suggested to play a modulatory role in microglial/macrophage activation, inflammation and tissue damage through the production of ROS, and is chronically up-regulated after SCI. NOX, microglia and astrocytes all show altered profiles with increased inflammatory morphology and basal activation states in both human and rodent aged populations. Thus, we hypothesize that age-related upregulation in NOX activity and expression leads to increased proinflammatory glial cell activation states, resulting in exaggerated glial responses and diminished recovery to spinal cord injury. To test this hypothesis, we propose three specific aims. In aim 1, we will characterize basal glial cell activation states and NOX expression and activity in the aging rodent spinal cord. In this aim, we will use double labeled immunohistochemistry and biochemical techniques to characterize NOX and glial cell activation state in 3 and 12 month aged rats. In aim 2, we will demonstrate the effects of aging on NOX expression and activity, glial cell activation, lesion size and functional recovery after spinal cod injury. In this aim, we will assess NOX and glial cell activation after a moderate contusion SCI in 3 and 12 month aged rats using immunohistochemistry, biochemistry, RT-qPCR and functional assessment. In aim 3, we will evaluate the effect of inhibition of NOX activity and expression on basal glial activation and response to injury. In this aim, we will assess the modulatory effects o NOX on microglia and astrocyte activation in aged rats using a NOX2 specific inhibitor. This effect will be investigated using immunohistochemistry, biochemistry, subcellular fractionation and functional assessment. The data generated in these aims will help to guide researchers and clinicians in more effective diagnosis and may provide important insight into potential future therapeutic targets for SCI in the aging population.

描述（由申请人提供）：目前美国有超过 250,000 人患有与脊髓损伤 (SCI) 相关的残疾。自 20 世纪 80 年代以来，老年人口中 SCI 的数量一直在稳步增加，并且与高发病率相关。衰老组织中氧化应激和活性氧 (ROS) 增加，并与中枢神经系统的组织损伤和慢性炎症有关。 (CNS). 这些应激源影响神经胶质细胞的活性，导致老年人群中出现的 SCI 延迟恢复，迄今为止还没有研究研究神经胶质细胞对 SCI 反应的年龄相关变化。酶被认为通过产生 ROS 在小胶质细胞/巨噬细胞激活、炎症和组织损伤中发挥调节作用，并且在 SCI、小胶质细胞和星形胶质细胞后长期上调。在人类和啮齿动物老年群体中，所有这些都显示出炎症形态和基础激活状态增加的变化，因此，我们认为与年龄相关的NOX活性和表达的上调导致促炎性神经胶质细胞激活状态增加，从而导致神经胶质细胞反应过度和减弱。为了检验这一假设，我们提出了三个具体目标，在目标 1 中，我们将表征衰老啮齿动物脊髓中的基底胶质细胞激活状态和 NOX 表达和活性。免疫组织化学和生化技术来表征 3 月龄和 12 月龄大鼠的 NOX 和神经胶质细胞激活状态在目标 2 中，我们将证明衰老对脊髓损伤后 NOX 表达和活性、神经胶质细胞激活、病变大小和功能恢复的影响。为此，我们将评估中度挫伤 SCI 后的 NOX 和神经胶质细胞活化情况。使用免疫组织化学、生物化学、RT-qPCR 和功能评估 3 个月和 12 个月大的大鼠。在目标 3 中，我们将评估抑制 NOX 活性和表达对基底胶质细胞活化和损伤反应的影响。使用 NOX2 特异性抑制剂对 NOX 对小胶质细胞和星形胶质细胞活化的调节作用将使用免疫组织化学、生物化学、亚细胞分级和功能进行研究。这些目标产生的数据将有助于指导研究人员和追随者进行更有效的诊断，并可能为老龄化人群中 SCI 的未来潜在治疗目标提供重要见解。