Oxidants, Mitochondria,and Renal Ischemia/Reperfusion

氧化剂、线粒体和肾缺血/再灌注

• 批准号: 6360025
• 负责人: LEE A MACMILLAN-CROW
• 金额: $ 24.4万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-09-01 至 2006-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6360025
• 关键词: antioxidants; cryopreservation; electron transport; enzyme activity; enzyme linked immunosorbent assay; free radical oxygen; high performance liquid chromatography; immunofluorescence technique; kidney function; kidney transplantation; laboratory rat; manganese; mitochondria; oxidative stress; oxidizing agents; renal ischemia /hypoxia; reperfusion; superoxide dismutase; tissue /cell culture; tissue /organ preservation; western blottings

This proposal addresses a critical issue in transplantation biology by focusing on the early injury (30 minutes to 2 days) following ischemic damage to the kidney during donor harvesting and/or preservation techniques, prior to transplantation. Mitochondrial dysfunction occurs during ischemia/reperfusion (I/R) episodes, and likely increases intra- mitochondrial reactive oxygen species (ROS) production. This study both extends and compliments the PI's in vitro and in vivo observations that during renal transplantation manganese superoxide dismutase (MnSOD), the major antioxidant in the mitochondria, is tyrosine nitrated and inactivated. The net result of loss of MnSOD activity is damage to sensitive electron transport systems and an accompanying increase in ROS production, thus amplifying further cellular injury. Preliminary studies using an in vivo rat kidney I/R model suggest that MnSOD is tyrosine nitrated early during renal I/R injury. These data are consistent with increased ROS-mediated injury during I/R and suggest that MnSOD is an early target that may result in oxidative injury and predispose the kidney to subsequent injury. We hypothesize that renal preservation and I/R leads to increased oxidative stress via alterations in MnSOD protein, which contributes to renal and mitochondrial injury prior to transplantation. The aims of this proposal are to determine the role that increased oxidant production has on the early events leading to renal dysfunction following preservation and I/R in vivo. Clearly, kidney damage occurs during I/R, therefore, strategies to limit the extent of renal damage during the cold and/or warm ischemic periods would most certainly improve graft function following renal transplantation. An underlying theme of these studies is to relate the increased ROS production to loss of MnSOD function during organ preservation and I/R. This information will be helpful in determining possible therapeutic interventions for improving current techniques in renal preservation and transplantation.

该提案通过关注移植前供体收获和/或保存技术期间肾脏缺血性损伤后的早期损伤（30分钟至2天）来解决移植生物学中的关键问题。 线粒体功能障碍发生在缺血/再灌注（I/R）发作期间，并且可能增加线粒体内活性氧（ROS）的产生。 本研究扩展并补充了PI的体外和体内观察结果，即在肾移植过程中，线粒体中的主要抗氧化剂锰超氧化物歧化酶（MnSOD）被酪氨酸硝化和灭活。 MnSOD活性丧失的最终结果是对敏感电子传递系统的损害以及随之而来的活性氧产生的增加，从而加剧了进一步的细胞损伤。 使用在体大鼠肾I/R模型的初步研究表明，MnSOD在肾I/R损伤的早期被酪氨酸硝化。 这些数据与I/R期间ROS介导的损伤增加一致，并表明MnSOD是可能导致氧化损伤并使肾脏易于发生后续损伤的早期靶点。我们推测，肾保护和I/R通过MnSOD蛋白的改变导致氧化应激增加，这有助于移植前肾和线粒体损伤。 本提案的目的是确定氧化剂产生增加对体内保存和I/R后导致肾功能障碍的早期事件的作用。 显然，肾损伤发生在I/R期间，因此，在冷缺血和/或热缺血期间限制肾损伤程度的策略肯定会改善肾移植后的移植物功能。 这些研究的一个基本主题是将器官保存和I/R期间增加的ROS产生与MnSOD功能丧失联系起来。 这些信息将有助于确定可能的治疗干预措施，以改善目前的技术在肾脏保存和移植。