Hepatic mitochondrial oxidative stress, AIDS and alcohol

描述（由申请人提供）：该项目阐明了亚细胞 酒精和酒精联合作用引起肝线粒体损伤的机制 核苷逆转录酶抑制剂（NRTI）用于艾滋病。 缺陷 线粒体（mt-）DNA复制，氧化应激（游离 自由基和抗氧化防御），以及肝微泡性脂肪变性 脂肪肝是NRTI、酒精肝和HIV感染的特征。 NRTI齐多夫定（3 '-叠氮基-2'，3 '-脱氧胸苷; AZT）消耗mtDNA， 引起微泡性肝脂肪变性（一种潜在的致命性肝病）， 和氧化应激。 根据其化学结构，NRTI三磷酸盐 竞争性抑制DNA pol-gamma（复制mtDNA的酶）或 混合动力学并耗尽线粒体DNA。 肝脂肪变性、mtDNA耗竭和 突变和酒精的结果。 HIV达特（反式激活因子） 肝线粒体谷胱甘肽（GSH），并有助于氧化应激。 酒精、艾滋病和NRTIs的联合作用尚不清楚，但 每一个都是合乎逻辑的结果。 工作假设指出：肝脏 NRTIs（艾滋病）和酒精联合造成的线粒体损伤是 比每个人都更糟糕。 其机制包括线粒体DNA复制缺陷， 突变、能量消耗和氧化应激。 转基因艾滋病小鼠 是专门的“生物工具”。 TG用酒精和NRTI治疗， 提出的实验。 表达HIV的靶向TG仅在 肝细胞（由白蛋白启动子驱动）精确定位肝特异性作用。 TGS 其普遍表达HIV达特（由B-肌动蛋白启动子驱动）， 系统性影响。 普遍表达NL 4 -3Agaglpol的TG是 系统性艾滋病毒。 酒精是通过配对喂养来管理的。 NRTI治疗 类似于临床上有用的。 生化、病理和 药理学研究解决了以下具体目的：1）定义 NRTIs引起的肝线粒体生物发生、功能和氧化应激 和酒精。 mtDNA、mtDNA、线粒体蛋白和GSH/GSSG减少， 预期8-OHdG增加和乌头酸酶失活; 2）确定 肝脂肪变性形态计量学（光和透射电子 显微镜）与酒精的NRTI治疗。 定量差异 肝细胞损伤（例如，改变的线粒体结构）。 综合效应比单独条件下的效应更差; 和3）减少艾滋病NRTIs和酒精对线粒体的损伤， 抗氧化剂或S-腺苷甲硫氨酸。 生物化学、分子和病理学 （以上）的变化得到改善。 这是一个“原则证明”。

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".