Hepatic mitochondrial oxidative stress, AIDS and alcohol

描述(由申请者提供)：本项目阐明亚细胞 酒精和乙醇联合作用对肝脏线粒体损伤机制的研究 治疗艾滋病的核苷逆转录酶抑制剂(NRTI)。有缺陷的 线粒体(mt-)DNA复制、氧化应激(自由之间的失衡 自由基和抗氧化剂防御)和肝脏微泡脂肪变性 (脂肪肝)是NRTIs、酒精性肝脏和艾滋病毒感染的特征。 NRTI齐多夫定(3‘-叠氮-2’，3‘-脱氧胸苷；AZT)耗尽线粒体DNA， 导致微泡性肝脏脂肪变性(一种潜在的致命肝病)， 和氧化应激。根据其化学结构，NRTI三磷酸盐 竞争性或用来抑制dna pol-Gamma(复制mtdna的酶)。 混合动力学和耗竭线粒体DNA。肝脏脂肪变性，线粒体DNA耗竭和 突变和酒精所致。HIV TAT(反式激活因子)耗尽 肝脏线粒体谷胱甘肽(GSH)与氧化应激有关。 酒精、艾滋病和NRTIs的联合作用尚不清楚，但来自 每一个都是合乎逻辑的结果。工作假说是：肝脏 NRTIs(用于艾滋病)和酒精的联合作用对线粒体的损害 每一次都比这更糟糕。机制包括线粒体DNA复制缺陷和 突变、能量耗竭和氧化应激。转基因艾滋病小鼠(TGS) 是专门的“生物工具”。在TGS中用酒精和NRTI治疗 拟议中的实验。靶向TGS独占表达HIV TAT 肝细胞(由白蛋白启动子驱动)精确定位肝脏特异性效应。TGS 普遍表达HIV Tat(由B-肌动蛋白启动子驱动)识别 系统性影响。无处不在表达NL4-3Agaglol的TGS是 全身性艾滋病毒。酒精是通过配对喂养的方式给药的。NRTI治疗 类似于临床上有用的。生化、病理和 药理学研究涉及以下具体目标：1)确定 NRTIs对肝脏线粒体生物发生、功能和氧化应激的影响 还有酒。线粒体DNA、线粒体RNA、线粒体蛋白和GSH/GSSG降低， 8-OHdG增加，乌头酸酶失活；2)定义 肝脏脂肪变性的形态计量学(光和透射电子 NRTI酒精治疗)。数量上的差异 观察到肝细胞损伤(如线粒体结构改变)。 综合影响比每一种情况下的效果都要差； 3)减少艾滋病NRTIs和酒精对线粒体的损害 抗氧化剂或S-腺苷蛋氨酸。生化、分子和病理学 (以上)更改已得到改进。这是一种“原则性证明”。

DESCRIPTION (provided by applicant): This project elucidates subcellular mechanisms of hepatic mitochondrial damage from the combination of alcohol and nucleoside reverse transcriptase inhibitors (NRTIs) for AIDS. Defective mitochondrial (mt-) DNA replication, oxidative stress (imbalance between free radicals and antioxidant defenses), and hepatic microvesicular steatosis (fatty liver) are features of NRTIs, the alcoholic liver, and HIV infection. The NRTI zidovudine (3'-azido-2',3'-deoxythymidine; AZT) depletes mtDNA, causes microvesicular hepatic steatosis (a potentially lethal liver disease), and oxidative stress. Based on their chemical structure, NRTI triphosphates inhibit DNA pol-gamma (the enzyme that replicates mtDNA) competitively or with mixed kinetics and deplete mtDNA. Hepatic steatosis, mtDNA depletion and mutation and result from alcohol. HIV tat (the transactivator) depletes hepatic mitochondrial glutathione (GSH) and contributes to oxidative stress. Combined effects of alcohol, AIDS and NRTIs are unknown, but potentiation from each is a logical outcome. The working hypothesis states: Hepatic mitochondrial damage from the combination of NRTIs (for AIDS) and alcohol is worse than that from each. Mechanisms include defective mtDNA replication and mutation, energy depletion, and oxidative stress. Transgenic AIDS mice (TGs) are specialized "biological tools". TGs are treated with alcohol and NRTIs in the proposed experiments. Targeted TGs that express HIV tat exclusively in hepatocytes (driven by albumin promoter) pinpoint liver-specific effects. TGs that ubiquitously express HIV tat (driven by B-actin promoter) identify systemic effects. TGs that express NL4-3Agaglpol ubiquitously are models of systemic HIV. Alcohol is administered by paired feeding. NRTI treatments resemble clinically useful ones. Biochemical, pathological and pharmacological studies address the following Specific Aims: 1) to define hepatic mitochondrial biogenesis, function, and oxidative stress from NRTIs and alcohol. Decreased mtDNA, mtRNA, mitochondrial proteins, and GSH/GSSG, increased 8-OHdG, and aconitase inactivation are expected; 2) to define hepatic steatosis morphometrically (light and transmission electron microscopy) in NRTI therapy with alcohol. Quantitative differences in hepatocyte damage (eg., altered mitochondrial structure) are observed. Combined effects are worse than those found with each individual condition; and 3) to reduce mitochondrial damage from AIDS NRTIs and alcohol using antioxidants or S-adenosylmethionine. Biochemical, molecular and pathological changes (above) are ameliorated. This serves as a "proof of principle".