Microdialysis Studies of Seizure-Induced Oxidative Stress

癫痫发作引起的氧化应激的微透析研究

• 批准号: 8420439
• 负责人: CRAIG E LUNTE
• 金额: $ 31.03万
• 依托单位: UNIVERSITY OF KANSAS LAWRENCE
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-07-16 至 2016-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8420439
• 关键词: 8-hydroxy-2' -deoxyguanosine; 8-hydroxyguanosine; Acids; Affect; Amino Acid Neurotransmitters; Antioxidants; Arachidonic Acids; Biochemical Pathway; Biological Markers; Brain; Brain region; Butyric Acids; Catecholamines; Chemicals; Coenzyme Q10; Convulsions; Coupled; DNA; DNA Damage; Disulfides; Dose; Epilepsy; Esters; Event; Experimental Models; Fatty Acids; Generalized seizures; Glutamates; Glutathione; Goals; Indomethacin; Kindling (Neurology); Laboratories; Lead; Lipid Peroxidation; Lipids; Malondialdehyde; Measures; Membrane; Metabolism; Methodology; Methods; Microdialysis; Modeling; Monitor; Neurologic; Nitrogen; Oxidative Stress; Oxygen; Pentylenetetrazole; Phospholipase A2; Plasma; Population; Principal Investigator; Prostaglandins; Proteins; Reactive Nitrogen Species; Reactive Oxygen Species; Research; Role; Sampling; Seizures; Status Epilepticus; Sulfhydryl Compounds; Techniques; Time; Tissue Sample; Urine; Wistar Rats; analytical method; buthionine; cyclooxygenase 1; diethyl maleate; gamma-Aminobutyric Acid; in vivo; insight; oxidation; oxidative damage; programs; research study; response; sample collection

DESCRIPTION (provided by applicant): The purpose of this project is to investigate the role of epileptic seizures in oxidative stress. There is a wealth of research that demonstrates that epileptic seizures lead to the formation of reactive oxygen and nitrogen species (ROS and RNS), which cause oxidative damage to DNA, lipids, and proteins. The role of excitotoxic events during epileptic seizures and the subsequent formation of ROS and RNS leading to oxidative stress will be investigated. Microdialysis sampling in discrete brain regions along with simultaneous recording of electrocorticographic (ECoG) activity will be used to probe chemical and electrographic activity changes in the brain associated with seizures. This will provide the ability to correlate biomarker levels with seizure activity. Microdialysis sampling will be used to continuously monitor several biochemical pathways prior to, during, and after induction of seizures. Microdialysis experiments will provide both temporal and spatial information about oxidative stress caused by seizures and the brains response to them. This approach will provide insight into questions that have remained unclear using plasma and urine sampling and standard tissue sampling techniques. Methods previously developed in the PI's laboratory will be used to monitor formation of ROS and RNS, biomarkers for DNA damage and lipid peroxidation, and the neurotransmitter amino acids and catecholamines. A new analytical method will be developed to detect prostanoids resulting from arachidonic acid metabolism. Three experimental models of epilepsy will be used in Wistar rats. The first two models use 3-mercaptopropionic acid (3-MPA) to chemically induce seizures in a controlled manner. For one model the 3-MPA is systemically administered and in the other it is locally administered through the microdialysis probe. A chemical kindling model will then be used, where a sub-threshold dose (for triggering seizures) of pentylenetetrazol will be used to generate seizures of increasing intensity, culminating in a generalized seizure convulsion. In addition, we propose to investigate the role of various modulation agents on the release of known oxidative stress agents during epileptic seizures. Using microdialysis sampling coupled to the advanced analytical methodology, the neurological events leading from seizures to oxidative stress will be investigated. In particular, the temporal relationship between oxidative stress (as measured by ROS and RNS formation), and neuroexcitation (as measured by the GABA/glutamate ratio and the catecholamines) will be determined and correlated to the duration and intensity of seizure episodes.

描述（由申请人提供）：本项目的目的是研究癫痫发作在氧化应激中的作用。有大量的研究表明，癫痫发作导致活性氧和氮物质（ROS和RNS）的形成，导致DNA，脂质和蛋白质的氧化损伤。将研究癫痫发作期间兴奋性毒性事件的作用以及随后导致氧化应激的ROS和RNS的形成。在离散脑区沿着进行微透析采样，同时记录皮质电图（ECoG）活动，将用于探测与癫痫发作相关的脑中化学和电图活动变化。这将提供将生物标志物水平与癫痫发作活动相关联的能力。微透析采样将用于 在诱导癫痫发作之前、期间和之后连续监测几种生化途径。微透析实验将提供有关癫痫发作引起的氧化应激和大脑对其反应的时间和空间信息。这种方法将提供对使用血浆和尿液采样以及标准组织采样技术仍不清楚的问题的深入了解。 PI实验室先前开发的方法将用于监测ROS和RNS的形成，这是DNA损伤和脂质过氧化的生物标志物，以及神经递质氨基酸和儿茶酚胺。将开发一种新的分析方法来检测花生四烯酸代谢产生的前列腺素。将在Wistar大鼠中使用三种癫痫实验模型。前两种模型使用3-巯基丙酸（3-MPA）以受控方式化学诱导癫痫发作。对于一种模型，3-MPA是全身给药，而在另一种模型中，它是通过微透析探针局部给药。然后将使用化学点燃模型，其中亚阈值剂量（用于触发癫痫发作）的戊四唑将用于产生增加的癫痫发作。 强度，最终导致全身性癫痫发作。此外，我们建议调查的作用，各种调制剂对释放已知的氧化应激剂在癫痫发作。使用微透析取样结合先进的分析方法，将研究从癫痫发作到氧化应激的神经事件。特别地，将确定氧化应激（如通过ROS和RNS形成测量的）和神经兴奋（如通过GABA/谷氨酸比率和儿茶酚胺测量的）之间的时间关系，并将其与癫痫发作的持续时间和强度相关联。