Nox Enzymes in Aging Bladder Dysfunction

Nox 酶在老化膀胱功能障碍中的作用

基本信息

批准号：
9214303
负责人：
Michael Raymond Ruggieri
金额：
$ 26.91万
依托单位：
TEMPLE UNIV OF THE COMMONWEALTH
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-02-15 至 2020-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9214303
关键词：
Adverse effects Age Aging Animals Bladder Bladder Diseases Bladder Dysfunction Bladder Tissue Calcium Chloride Channels Connexins Data Dependence Disease Enzyme Inhibitor Drugs Enzyme Reactivation Enzymes Exocytosis Functional disorder Generations Human Hydrogen Peroxide Inflammatory Knockout Mice Mediating Mediator of activation protein Mitochondria Mus Muscle Contraction Muscle function NADPH Oxidase Overactive Bladder Oxidases Oxidation-Reduction Oxidative Stress Oxygenases Pathogenesis Pathologic Pathway interactions Phosphotransferases Physiology Predisposing Factor Process Protein Isoforms Proteins Reactive Oxygen Species Regulation Role Sensory Signal Pathway Signal Transduction Signaling Molecule Smooth Muscle Source Specificity Stimulus Stress Structure Superoxides Therapeutic Intervention Urothelial Cell Urothelium age related aging population cell type experimental study improved inhibitor/antagonist interest middle age novel oxidative damage prototype public health relevance receptor response therapeutic target

项目摘要

DESCRIPTION (provided by applicant): Specific Aim 1. To identify the enzymatic sources of ROS generation and Nox expression in the urothelial and bladder tissues at different ages (young vs. middle-aged and aging) in experimental animals (mice) and humans. This aim will identify the origin of ROS and specific contribution from Nox enzymes, and then the expression of the Nox in the urothelium and smooth muscle, and the age dependence. These form the basis for the action of Nox-derived ROS in the bladder. 1. Major sources for ROS generation in urothelium and smooth muscle (NADPH oxidases; mitochondria; other oxidases/oxygenases) 2. Expression and activity of Nox2 (prototype) and other Nox isoforms (Nox1, Nox4) in urothelium and smooth muscle 3. Age-dependent changes to ROS generation and Nox activities. Specific Aim 2. To determine age-associated Nox activation in urothelium and consequent oxidative stress damage of bladder function at different ages using wild-type and Nox knockout mice. This aim will define Nox activation in the urothelium in response to the inflammatory factors encountered during aging, its consequent action on urothelial and smooth muscle function and the underlying mechanisms. 1. To identify the changes in urothelial Nox activity in response to inflammatory stimuli in young versus middle-aged and aging bladders 2. To characterize the effects of Nox activation on urothelial ATP release and smooth muscle contractions in young versus aging bladders 3. To discover the key signaling mediators and proteins in aging-dependent Nox activation in urothelial and bladder tissues. Specific Aim 3. To investigate the cellular mechanisms of redox-regulation in urothelial function using primary urothelial cells isolated from wild-type versus Nox knockout mice at different ages. This aim will explore further the cellular mechanisms of ROS/Nox regulation and their interaction with the key pathways for aging and bladder function, providing fundamental mechanisms for Nox contribution to bladder dysfunction during aging. 1. Effect of different ROS enzyme inhibitors and ROS scavengers on urothelial cell release of ATP and other sensory mediators. 2. Changes in urothelial Nox activity in response to inflammatory stimuli. Focus on the stress pathway. 3. Effects of ROS (H2O2) at different levels on urothelial ATP release and calcium release. 4. Importance of ROS for downstream intracellular signaling pathways in the regulation of urothelial transmitter release - role of intracellular Ca2+, exocytosis, chloride channels, kinases and connexins.

描述（由适用提供）：特定目的1。确定在不同年龄（年轻人与衰老）中，在实验动物（小鼠）和人类中，尿路上皮和膀胱组织中ROS的产生和NOX表达的酶促来源。该目标将确定ROS的起源和NOX酶的特定贡献，然后确定NOX在尿皮上和平滑肌中的表达以及年龄的依赖性。这些构成了NOX衍生ROS在膀胱中作用的基础。 1。尿路上皮和平滑肌中ROS产生的主要来源（NADPH氧化酶；线粒体；其他氧化酶/氧酶） 2。尿皮细胞和平滑肌中NOX2（原型）和其他NOX同工型（NOX1，NOX4）的表达和活性 3。与年龄相关的ROS生成和NOX活动的变化。具体目标2。确定使用野生型和NOX敲除小鼠在不同年龄在不同年龄的尿路上皮中与年龄相关的NOX激活。该目标将根据衰老过程中遇到的炎症因素，其对尿路上皮和平滑肌功能的作用以及基本机制的作用来定义尿路上皮中的NOx激活。 1。确定年轻与中年和衰老的炎症刺激的尿路上皮活性的变化 2。表征NOX激活对尿路上皮ATP释放的影响和年轻人与衰老的肌肉收缩的影响 3。发现尿路上皮和膀胱组织中衰老依赖性NOX激活中的关键信号介质和蛋白质。具体目的3。使用从野生型与NOX敲除小鼠分离的原代尿路上皮细胞在不同年龄分离的尿路上皮功能中氧化还原调节的细胞机制。该目标将进一步探索ROS/NOX调节的细胞机制及其与衰老和膀胱功能的关键途径的相互作用，从而为NOX在衰老过程中提供了NOX贡献的基本机制。 1。不同ROS酶抑制剂和ROS清除剂对ATP和其他感觉介质的尿路上皮细胞释放的影响。 2。响应炎症刺激的尿路上皮NOX活性的变化。专注于应力途径。 3。ROS（H2O2）在不同水平上对尿路上皮ATP释放和钙释放的影响。 4。在调节尿路上皮发射机释放的调节中，ROS对下游细胞内信号通路的重要性 - 细胞内Ca2+的作用，胞吐作用，氯化物通道，激酶和连接素。