Oxidative Stress Responses to Loss and Recovery of Sleep

描述（由申请人提供）：被广泛接受的两个概念是：睡眠剥夺损害健康和睡眠恢复具有动态治疗能力。提出这项研究的基本原理是，尚未确定特定组织在睡眠剥夺后需要修复或通过睡眠恢复“恢复”。我们的长期目标是提供构成睡眠和睡眠缺失特性的生理过程和结果的切实证据。在动物模型中，睡眠剥夺会产生一种最终变得高度致命的疾病，缺乏特定的定位，并且随着睡眠而可逆，这意味着通过生化过程或功能异常进行调解。最近在睡眠不足的大鼠身上的发现提供了证据，证明氧化应激和抗氧化剂消耗是缺失的生物中介。本提案的目的是确定氧化应激相关损伤的目标，并确定抗氧化剂消耗和细胞损伤对生理和临床体征的影响程度。核心假设是，抗氧化状态的降低导致多种调节系统中可修复和不可修复的细胞损伤的广泛增加，最终导致补偿不足和病理生理体征的发展。初步数据支持这一假说的形成，提供了睡眠不足导致的氧化应激下肝组织DNA损伤和细胞凋亡增加的证据。直到再次允许睡眠，抗氧化酶才会做出补偿反应。在Aim 1下的实验中，将在大鼠睡眠缺失和恢复期间确定氧化应激对细胞脂质、蛋白质和DNA靶点的损伤以及对特定器官系统的损伤定位。Aim 2下的实验将确定导致细胞死亡的细胞损伤程度，以及在睡眠缺失和恢复期间增加细胞修复和更新的程度。Aim 3下的研究将测试在多大程度上操纵抗氧化状态可以改变睡眠剥夺引起的细胞损伤和生理体征。拟议的研究是合作的，整合了睡眠生理学和自由基生物学的专业知识。研究设计包括有计划地比较不同时间的睡眠剥夺、睡眠限制和控制条件。分析包括生化、免疫测定和免疫组织化学方法。提出的研究的意义是推进医学干预，促进睡眠的恢复功能。

DESCRIPTION (provided by applicant): Widely accepted are two notions: sleep deprivation impairs health and sleep recovery has dynamic healing powers. The rationale for the proposed research is that specific tissues have not yet been identified as being in need of repair after sleep deprivation or "restored" by sleep recovery. Our long-term goal is to provide tangible evidence of physiological processes and outcomes that compose the properties of sleep and sleep loss. Sleep deprivation in the animal model produces a condition that eventually becomes highly lethal, lacks specific localization, and is reversible with sleep, implying mediation by a biochemical process or functional abnormality. Recent findings in sleep-deprived rats have provided evidence that the missing biological mediation is oxidative stress and antioxidant depletion. The objectives of this proposal are to identify targets of oxidative stress-associated damage and to determine the extent to which antioxidant depletion and cell damage affect physiological and clinical signs. The central hypothesis is that reductions in antioxidant status cause widespread increases in repairable and irreparable cell damage within multiple regulatory systems, leading ultimately to inadequate compensation and to the development of pathophysiological signs. Preliminary data support the formulation of this hypothesis by providing evidence of increased DNA damage and apoptosis in hepatic tissue undergoing oxidative stress induced by sleep loss. Antioxidant enzymes fail to respond in compensation until sleep is again permitted. In experiments under Aim 1, damage to cellular lipid, protein, and DNA targets of oxidative stress and localization of damage to specific organ-systems will be determined in rats during sleep loss and recovery. Experiments under Aim 2 will determine the extent of cell injury leading to cell death and to increased cell repair and renewal during sleep loss and recovery. Studies under Aim 3 will test the extent to which manipulation of antioxidant status can change sleep deprivation-induced cell damage and physiological signs. The proposed research is collaborative and integrates expertise in sleep physiology and free radical biology. The research design is composed of planned comparisons of different durations of sleep deprivation, sleep restriction, and control conditions. Analyses include biochemical, immunoassay, and immunohistochemical methods. The significance of the proposed research is advancement of medical interventions that promote the restorative functions of sleep.