INDUCIBLE DNA REPAIR IN CEREBRAL ISCHEMIA

脑缺血中的诱导性 DNA 修复

• 批准号: 2685783
• 负责人: Jun Chen
• 金额: $ 17.43万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-06-16 至 2001-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2685783
• 关键词: DNA damage; DNA directed DNA polymerase; DNA repair; cell death; cell type; cerebral artery; cerebral ischemia /hypoxia; enzyme activity; enzyme inhibitors; gene expression; histology; laboratory rat; neurons; phosphoester ligase; plasmids; proliferating cell nuclear antigen; tissue /cell culture

DESCRIPTION (Adapted from Applicant's Abstract): Recent studies have implicated oxidative DNA damage in mediating neuronal death in cerebral ischemia and in neurodegenerative diseases. Little is known about how neuronal DNA repair mechanisms respond to ischemic injury and oxidative DNA damage. The broad, long-term objective of this project is to identify the role of inducible DNA excision repair in the regulation of cell death from acute ischemic brain injury. The specific objectives are: (1) Determine inducible DNA excision repair activity in an in vivo rat model of focal ischemia and reperfusion. (2) Determine inducible DNA excision repair activity in mechanist-ically defined in vitro models of neuronal ischemia and neuronal cell death. (3) Determine activity of selected DNA repair enzymes and expression of DNA repair genes in vivo focal cerebral ischemia and in vitro cell culture models of ischemia and neuronal cell death. (4) Determine the role of selective DNA repair enzymes in inducible DNA excision repair in focal ischemia and in vitro models of ischemia and neuronal death, by altering the activity of these enzymes. in vitro models allow for precise mechanistic studies to be performed, but the complex inter-dependent pathophysiologic changes that occur after ischemia in vivo cannot be completely modeled. Accordingly, both primary cortical neuronal cultures and rat temporary middle cerebral artery occlusion models will be used. Methods for detection of single, double and total DNA breaks will be used to study the evolution of DNA damage in these models. The ability of whole-cell extracts to repair damage to plasmid DNA by base excision repair or nucleotide excision repair will be used to assay DNA repair activity in vivo and in vitro. The expression and activity of DNA repair enzymes will be studied and the effect of pharmacologic inhibition of these enzymes upon DNA damage and neuronal survival investigated.

描述（改编自申请人的摘要）：最近的研究 氧化性DNA损伤介导脑内神经元死亡 局部缺血和神经变性疾病。 很少有人知道如何 神经元DNA修复机制对缺血损伤和氧化DNA应答 损害 该项目的广泛、长期目标是确定 诱导型DNA切除修复在细胞死亡调控中的作用 急性缺血性脑损伤 具体目标是：（1）确定 在体内大鼠局灶性脑缺血模型中可诱导DNA切除修复活性 缺血和再灌注。 (2)确定诱导型DNA切除修复 在神经元缺血的机械定义的体外模型中的活性 和神经细胞死亡。 (3)确定选定的DNA修复活性 在体局灶性脑缺血中的酶和DNA修复基因的表达 以及缺血和神经元细胞死亡的体外细胞培养模型。 （四） 确定选择性DNA修复酶在诱导型DNA切除中的作用 在局部缺血和缺血和神经元死亡的体外模型中的修复， 通过改变这些酶的活性。 体外模型允许 精确的机制研究，但复杂的相互依赖 体内局部缺血后发生的病理生理变化不能 完全模仿。 因此，两种原代皮层神经元培养物 采用大鼠大脑中动脉暂时闭塞模型。 检测单、双和总DNA断裂的方法将用于 研究这些模型中DNA损伤的演变。 的能力 全细胞提取物通过碱基切除修复修复质粒DNA损伤 或核苷酸切除修复将用于测定DNA修复活性， 体内和体外。 DNA修复酶的表达和活性将 这些酶的药理学抑制作用， 研究DNA损伤和神经元存活。