Effects of Age on Ethanol Withdrawal Toxicity: Mechanisms and Therapy

年龄对乙醇戒断毒性的影响：机制和治疗

基本信息

批准号：
7147620
负责人：
Marianna E Jung
金额：
$ 27.81万
依托单位：
UNIVERSITY OF NORTH TEXAS HLTH SCI CTR
依托单位国家：
美国
项目类别：
财政年份：
2006
资助国家：
美国
起止时间：
2006-08-15 至 2011-07-31
项目状态：
已结题

项目摘要

DESCRIPTION (provided by applicant): Alcoholism is a consequence of prolonged ethanol abuse and has been associated with the aging of cognition and psychomotor function. Because alcoholics go through repeated rounds of ethanol withdrawal (EW), we propose using repeated EW to investigate whether aging with exacerbates the effects of EW on cognition and psychomotor function through oxidative insults to mitochondria. We also propose to investigate whether a loss of estrogen contributes to this problem. This work may have particular significance for female alcoholics, who simultaneously undergo EW and estrogen deficiency in their advanced age. Our pilot study has demonstrated that estrogen protects against EW toxicity in both young and old female rats. In the proposed study, we will use three age groups of ovary-intact female rats: 5 months old, 12 months old, and 16 months old, when the ethanol diet begins. We propose to use a diet cycle of 25 days of ethanol followed by 5 days of withdrawal repeated for 5 cycles. These cycles will be timed such that rats will be at the estrus phase (high estrogen levels) when diet is removed and EW signs peak. In Aim 1, we will determine the effects of EW on cognition and psychomotor functions during aging. Rats at the estrus phase will be withdrawn from an intermittent ethanol diet and tested for psychomotor function using established behavioral methods. In Aim 2, we will characterize the mitochondrial oxidative mechanisms underlying the deleterious interaction between age and EW. We will measure oxidative markers (reactive oxygen species, lipid peroxidation) and consequent mitochondrial dysfunction (collapse of mitochondrial membrane potential and ATP loss). In Aim 3, we will characterize oxidatively modified mitochondrial enzyme cytochrome c oxidase (COX) which we hypothesize mediates the deleterious interaction between age and EW. We will quantify carbonylation of COX and lipid-peroxide-modified COX using liquid chromatography-mass spectrometry and tandem mass spectrometry. In Aim 4, we will determine whether estrogen deficiency during aging contributes to EW-induced oxidative damage to mitochondria and psychomotor deficit. The same psychomotor and oxidative markers will be measured in ovary-intact and ovariectomized rats with or without estrogen replacement. These studies may ultimately contribute to a better understanding of and strategy for female alcoholics undergoing EW during brain aging.

描述（由申请人提供）：酗酒是长期滥用乙醇的结果，与认知和精神运动功能的老化有关。由于酗酒者会经历反复的乙醇戒断（EW），因此我们建议使用重复的乙醇戒断（EW）来研究衰老是否会通过对线粒体的氧化损伤而加剧乙醇戒断对认知和精神运动功能的影响。我们还建议调查雌激素的丧失是否会导致这个问题。这项工作对于女性酗酒者可能具有特别的意义，因为她们在高龄时同时患有电子酒和雌激素缺乏症。我们的初步研究表明，雌激素可以防止年轻和年老雌性大鼠的电子战毒性。在拟议的研究中，我们将使用三个年龄组的卵巢完整的雌性大鼠：5 个月大、12 个月大和 16 个月大（当乙醇饮食开始时）。我们建议使用 25 天乙醇饮食周期，然后 5 天戒断，重复 5 个周期。这些周期的时间安排应使大鼠处于发情期（雌激素水平高），此时停止饮食且 EW 迹象达到顶峰。在目标 1 中，我们将确定电子波对衰老过程中认知和精神运动功能的影响。发情期的大鼠将停止间歇性乙醇饮食，并使用既定的行为方法测试精神运动功能。在目标 2 中，我们将描述年龄与 EW 之间有害相互作用背后的线粒体氧化机制。我们将测量氧化标记（活性氧、脂质过氧化）和随之而来的线粒体功能障碍（线粒体膜电位崩溃和 ATP 损失）。在目标 3 中，我们将表征氧化修饰的线粒体酶细胞色素 C 氧化酶 (COX)，我们假设它介导年龄和 EW 之间的有害相互作用。我们将使用液相色谱-质谱法和串联质谱法定量 COX 和脂质过氧化物修饰的 COX 的羰基化。在目标 4 中，我们将确定衰老过程中雌激素缺乏是否会导致电波诱导的线粒体氧化损伤和精神运动缺陷。在使用或不使用雌激素替代的卵巢完整和卵巢切除的大鼠中，将测量相同的精神运动和氧化标记物。这些研究最终可能有助于更好地理解女性酗酒者在大脑衰老过程中接受电子戒酒的策略并制定策略。