Aging, Reperfusion, and Apoptosis:A Proteasome Approach

衰老、再灌注和细胞凋亡：蛋白酶体方法

• 批准号: 6625766
• 负责人: LUKE I. SZWEDA
• 金额: $ 34.28万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-05-01 至 2005-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6625766
• 关键词: BCL2 gene /protein; Bax gene /protein; SDS polyacrylamide gel electrophoresis; age difference; aging; apoptosis; coronary occlusion /thrombosis; cytochrome c; densitometry; electron microscopy; electrospray ionization mass spectrometry; free radicals; high performance liquid chromatography; laboratory rat; matrix assisted laser desorption ionization; monoclonal antibody; proteasome; reperfusion; western blottings

DESCRIPTION: (provided by applicant) Complications arising from reduction of blood supply to the heart are a leading cause of death and debilitation worldwide. Depending on the duration and severity of the ischemic event, irreparable alterations in cellular homeostasis lead to necrotic cell death. However, reperfusion of viable cardiac tissue can result in cell death by processes unrelated to necrosis. This has been attributed, in part, to increases in the production of oxygen radicals and the induction of programmed cell death, termed apoptosis. The proteasome, a major intracellular proteolytic system, appears to play a critical role in the prevention of apoptosis by degrading certain pro-apoptotic factors. Nevertheless, the proteasome is itself modified by free radical processes and exhibits dramatic declines in activity as a result of coronary occlusion/reperfusion. Furthermore, proteasome expression and specific activity decrease as a function of age. The objectives of the proposed research are to establish a mechanistic link between age and free radical induced loss in proteasome function and stimulation of the apoptotic process during coronary occlusion/reperfusion. Utilizing a physiologically relevant in vivo rat model of coronary occlusion and reperfusion this study seeks to: 1) Identify alterations in proteasome activity; 2) Defme free radical processes which result in loss in proteasome activity; 3) Characterize the progression of apoptosis; and 4) Establish mechanisms by which proteasome inactivation enhances apoptosis. In each aim, the effects of duration of occlusion and reperfusion and age of the animal will be assessed. Thus, these studies will establish biochemical mechanisms by which proteasome function is altered and when, in the sequence of coronary occlusion and reperfusion, critical oxidative events occur. Furthermore, the role of proteasome inactivation in the induction of apoptosis will be established. Finally, the contribution of age-dependent declines in proteasome activity to the progression and extent of coronary occlusion/reperfusion induced apoptosis will be elucidated. Results of the proposed study will therefore define molecular events which are likely to impact the long term progression of heart disease and indicate efficient strategies for favorably influencing the outcome.

描述：（由申请人提供）减少的并发症 心脏的血液供应是死亡和虚弱的主要原因 全世界。根据缺血事件的持续时间和严重程度， 细胞稳态无法弥补的改变导致坏死细胞死亡。 但是，可行心脏组织的再灌注可能会导致细胞死亡 与坏死无关的过程。这部分归因于 氧自由基的产生和程序的诱导增加 细胞死亡，称为凋亡。蛋白酶体，一种主要的细胞内蛋白水解 系统，似乎在预防细胞凋亡中起着至关重要的作用 降解某些促凋亡因素。然而，蛋白酶体本身是 通过自由基过程和展示活动的急剧下降而修改 由于冠状动脉闭塞/再灌注。此外，蛋白酶体 表达和特定活性随着年龄的函数而降低。目标 拟议的研究是建立年龄和年龄之间的机械联系 自由基诱导的蛋白酶体功能丧失和刺激 冠状动脉闭塞/再灌注过程中的凋亡过程。利用一个 冠状动脉闭塞的生理相关体内大鼠模型 再灌注本研究试图：1）确定蛋白酶体的变化 活动; 2）DEFME自由基过程，导致蛋白酶体损失 活动; 3）表征凋亡的进展； 4）建立 蛋白酶体失活增强凋亡的机制。在每个目标中， 闭塞和再灌注持续时间以及动物的年龄的影响将 被评估。因此，这些研究将建立生化机制 蛋白酶体功能会改变，何时在冠状动脉闭塞 和再灌注，发生关键的氧化事件。此外， 蛋白酶体在诱导凋亡的诱导中将被建立。 最后，蛋白酶体活动的年龄依赖性下降对 冠状动脉闭塞/再灌注的进展和程度诱导凋亡 将阐明。因此，拟议的研究的结果将定义 可能影响心脏长期进展的分子事件 疾病并指出有效影响的有效策略 结果。