Role of Dual Oxidase in post-stroke brain inflammation and injury

双氧化酶在中风后脑炎症和损伤中的作用

基本信息

批准号：
10214199
负责人：
UMADEVI V WESLEY
金额：
$ 15.55万
依托单位：
UNIVERSITY OF WISCONSIN-MADISON
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-04-01 至 2023-09-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10214199
关键词：
Acute Address Animal Model Animals Apoptosis Apoptotic Biochemical Blood Vessels Brain Brain Infarction Brain Injuries Brain Ischemia Calcium Signaling Caregivers Cell Culture Techniques Cell Survival Cell physiology Cells Cerebral Ischemia Cerebrovascular Disorders Cerebrum Cessation of life Data Development Dinucleoside Phosphates Encephalitis Endothelium Enzymes Epithelial Cells Event Excision Family Free Radicals Functional disorder Future Gene Expression Genetic Glucose Goals Grant Host Defense Hydrogen Peroxide Hypertension In Vitro Inflammation Inflammatory Injury Ischemia Ischemic Brain Injury Ischemic Stroke Knock-out Lead Link Malignant Neoplasms Mediating Modeling Motor NADPH Oxidase Neurologic Neurologic Dysfunctions Neurons Niacinamide Output Oxidants Oxidases Oxidative Stress Oxygen Pathologic Patient Care Phase Physiological Prevalence Process Production Proteins RNA Reactive Inhibition Reactive Oxygen Species Recovery of Function Reperfusion Injury Reperfusion Therapy Rodent Role Sensory Signaling Molecule Small Interfering RNA Stroke Superoxides Therapeutic Studies Thyroid Gland Thyroid Hormones Time Tissues Vascular Diseases Work airway epithelium antimicrobial blood-brain barrier disruption brain endothelial cell brain repair burden of illness cell motility cerebral ischemic injury cerebrovascular cognitive function cytokine deprivation disability endothelial dysfunction functional outcomes improved in vivo injury and repair inorganic phosphate member molecular array neuroinflammation neuron loss new therapeutic target novel novel strategies overexpression post stroke public health relevance repaired spatiotemporal stroke model stroke outcome stroke patient success therapeutic evaluation treatment strategy

Acute Address Animal Model Animals Apoptosis Apoptotic Biochemical Blood Vessels Brain Brain Infarction Brain Injuries Brain Ischemia Calcium Signaling Caregivers Cell Culture Techniques Cell Survival Cell physiology Cells Cerebral Ischemia Cerebrovascular Disorders Cerebrum Cessation of life Data Development Dinucleoside Phosphates Encephalitis Endothelium Enzymes Epithelial Cells Event Excision Family Free Radicals Functional disorder Future Gene Expression Genetic Glucose Goals Grant Host Defense Hydrogen Peroxide Hypertension In Vitro Inflammation Inflammatory Injury Ischemia Ischemic Brain Injury Ischemic Stroke Knock-out Lead Link Malignant Neoplasms Mediating Modeling Motor NADPH Oxidase Neurologic Neurologic Dysfunctions Neurons Niacinamide Output Oxidants Oxidases Oxidative Stress Oxygen Pathologic Patient Care Phase Physiological Prevalence Process Production Proteins RNA Reactive Inhibition Reactive Oxygen Species Recovery of Function Reperfusion Injury Reperfusion Therapy Rodent Role Sensory Signaling Molecule Small Interfering RNA Stroke Superoxides Therapeutic Studies Thyroid Gland Thyroid Hormones Time Tissues Vascular Diseases Work airway epithelium antimicrobial blood-brain barrier disruption brain endothelial cell brain repair burden of illness cell motility cerebral ischemic injury cerebrovascular cognitive function cytokine deprivation disability endothelial dysfunction functional outcomes improved in vivo injury and repair inorganic phosphate member molecular array neuroinflammation neuron loss new therapeutic target novel novel strategies overexpression post stroke public health relevance repaired spatiotemporal stroke model stroke outcome stroke patient success therapeutic evaluation treatment strategy

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项目摘要

Project Summary Ischemic brain damage remains a leading cause of long-term disability and death with limited treatment options. Cerebral ischemic injury is strongly associated with excessive production of reactive oxygen species (ROS) that contributes to endothelial dysfunction, blood brain barrier disruption, neuronal cell death, and worsened brain damage. Thus, efforts to curtail ROS have major impact on improving stroke outcome. Dual oxidases (Duox) are novel members of reduced nicotinamide dinucleotide phosphate oxidases family. The main function of Duox1 is to generate hydrogen peroxide (H2O2)/ROS. Duox1 at low levels, is involved in essential cellular functions, antimicrobial host defense, thyroid hormone production, and airway epithelial cell migration, and injury. However, excessive production and activation of Duox1 may contribute to pathological events including inflammation, apoptosis, hypertension, cancer, and tissue damage. The direct link between Duox1 and ROS in airway epithelial cells was shown in our previous work. Pro-inflammatory cytokines and deranged calcium signaling increase the activity and expression of Duox in airway epithelial cells and thyroid tissues. Interestingly, ischemic stroke causes aberrant Ca2+ influx. Despite these compelling observations, the specific roles of Duox in the brain and cerebral ischemia are largely unknown. We have recently identified that focal cerebral ischemia in rodents, and in-vitro oxygen glucose deprivation rapidly induce the expression of Duox1 in endothelial and neuronal cells in association with increased ROS production. However, pre-treatment of neuronal cells with Duox1 specific small interfering RNA decreased Duox1 expression and ROS levels. These data led us to hypothesize that cerebral ischemia evokes Duox1 over-expression which in turn increases ROS in the brain, leading to exacerbation of pro-inflammatory and apoptotic processes that worsen the brain injury. We further propose that Duox1 inhibition has a great potential to mitigate post-ischemic brain damage and neurological dysfunction. Accordingly, our specific Aims are; To determine the spatiotemporal changes in the expression of Duox1 in brain following focal cerebral ischemia/reperfusion; To determine if Duox1 inhibition or genetic loss of Duox1 decreases ROS, and reduces ischemic brain damage, and thus improves post-stroke functional recovery; To investigate the role of Duox1 as a key driver of inflammatory and apoptotic processes in ischemic brain. We will address these Aims using a wide array of molecular, cellular, and biochemical approaches in both in vivo animal, and in vitro cell culture models. Overall, this ‘proof of concept’ study will determine a previously unidentified role for Duox1 in ischemic brain. These studies pave the way towards better understanding of the role of Duox1 mediated ROS and neuro-inflammatory mechanisms in brain and may open up a promising new approach for treating cerebro- vascular diseases.

项目摘要缺血性脑损伤仍然是长期残疾和死亡的主要原因，并有限的治疗选择。脑缺血性损伤与活性氧（ROS）的产生密切相关，该物种的产生导致内皮功能障碍，血脑屏障破坏，神经元细胞死亡和大脑恶化损害。那，减少ROS的努力对改善中风结果产生了重大影响。双氧化物（Duox）是减少烟酰胺二核苷酸磷酸氧化物家族的新成员。 Duox1的主要功能是生成过氧化氢（H2O2）/ROS。 DUOX1低水平，参与必需的细胞功能，抗微生物宿主防御，甲状腺荷鸟生产和气道上皮细胞迁移以及受伤。然而， DUOX1的过度产生和激活可能导致病理事件，包括炎症，凋亡，高血压，癌症和组织损伤。 DUOX1和ROS之间的直接连接细胞在我们以前的工作中显示。促炎性细胞因子和教钙信号传导增加了 DUOX在气道上皮细胞和甲状腺组织中的活性和表达。有趣的是，缺血性中风原因异常CA2+影响。尽管有这些令人信服的观察，但Duox在大脑和大脑中的特定作用缺血在很大程度上未知。我们最近确定啮齿动物中的局灶性脑缺血和体外缺血氧气剥夺迅速诱导DUOX1在内皮细胞和神经元细胞中的表达与ROS产生增加相关。但是，用Duox1特异性小的神经元细胞进行预处理干扰RNA恶化DUOX1表达和ROS水平。这些数据导致我们假设大脑缺血唤起Duox1的过表达，从而增加大脑中的ROS，导致加剧促炎和凋亡过程会使脑损伤恶化。我们进一步建议DUOX1抑制减轻缺血后脑损伤和神经功能障碍具有很大的潜力。根据我们的说法具体目标是；确定局灶性后DUOX1表达的时空变化脑缺血/再灌注；为了确定duox1抑制或duox1的遗传丧失是否会降低ROS，并且减少缺血性脑损伤，从而改善中风后功能恢复；调查 DUOX1是缺血性大脑中炎症和凋亡过程的关键驱动力。我们将解决这些目标在体内动物和体外细胞中使用多种分子，细胞和生化方法文化模型。总体而言，这项“概念证明”研究将确定duox1先前未确定的角色缺血性大脑。这些研究为更好地理解DUOX1介导的ROS的作用铺平了道路和大脑中的神经炎症机制，并可能为治疗脑治疗的新方法打开血管疾病。