Thiol Redox Modulation of NF-kB Pathway in Alcoholic Liver Injury.

酒精性肝损伤中 NF-kB 通路的硫醇氧化还原调节。

• 批准号: 8055022
• 负责人: DERICK S HAN
• 金额: $ 14.45万
• 依托单位: UNIVERSITY OF SOUTHERN CALIFORNIA
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-04-10 至 2013-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8055022
• 关键词: Accounting; Affect; Alcohol consumption; Alcohol-Induced Disorders; Alcoholic Liver Diseases; Alcoholic liver damage; Alcohols; Antioxidants; Apoptosis; Apoptotic; Area; Binding; Biochemical; Biological Models; Biology; Cell Death; Cell Nucleus; Cells; Chronic; Complex; Confocal Microscopy; Cysteine; Cytoplasm; DNA; Data; Disease; Flow Cytometry; Free Will; Funding; Future; Gene Expression; Generations; Genes; Genetic Transcription; Glutathione; Glutathione Disulfide; Goals; Grant; Hepatic; Hepatocyte; Hepatotoxicity; Heterogeneity; Imaging Techniques; Immunoblotting; Inflammation; Infusion procedures; Injury; Investigation; Knockout Mice; Kupffer Cells; Laboratories; Light; Link; Liver; Liver diseases; Mediating; Methods; Mitochondria; Modeling; Molecular Biology; Molecular Biology Techniques; Mus; NF-kappa B; Necrosis; Nuclear; Outcome Study; Oxidants; Oxidation-Reduction; PTEN gene; Pathogenesis; Pathway interactions; Population; Prevention; Proteins; Publications; Reactive Oxygen Species; Research; Research Personnel; Research Proposals; Resistance; Science; Signal Pathway; Signal Transduction; Sulfhydryl Compounds; Techniques; Testing; Thioctic Acid; Time; Transcriptional Activation; Tumor Necrosis Factor-alpha; Wages; Work; alcohol abuse therapy; alcohol research; base; cell injury; cell transformation; cellular imaging; chronic alcohol ingestion; cytokine; cytotoxic; experience; feeding; in vivo; injured; insight; intercellular communication; novel therapeutics; oxidation; public health relevance; receptor; transcription factor

DESCRIPTION (provided by applicant): It is my long term goal to become an independent investigator and study the pathogenesis of liver disease including alcoholic liver disease. Obtaining a K01 grant would be an important step for me to become an independent researcher since it will provide me an opportunity and time to overcome many weaknesses (lack of preliminary data and publications in alcohol research, previous lack of focus on a clinically important model system, inexperience in cell signaling, cell imaging, and molecular biology techniques) in my background to become competitive for an R01 in alcohol research in the near future. By providing my salary and research funding, the K01 grant will free me from working on other PIs' projects and allow me to really focus on alcohol, as well as become more experienced in cell signaling, cell imaging, and molecular biology. The area of science I have chosen to pursue in the upcoming period is at the interface of redox biology, signal transduction and hepatotoxicity using alcohol as a model of liver injury. TNF-induced damage to hepatocytes remains a central point in alcohol-induced liver damage. Recent evidence from our laboratory suggests that redox alterations through GSH modulation or oxidant treatment sensitize hepatocytes to TNF-induced apoptosis by inhibiting NF-?B transcriptional activity. This suggests a link between TNF secretion and redox alterations in mediating damage to liver. The hypothesis to be tested is that redox alterations caused by alcohol sensitize hepatocytes to TNF-induced apoptosis through inhibition of NF-?B dependent signaling pathways. The hypothesis will be tested with the following specific aims: 1) Determine how alcohol-induced alterations of cellular redox status modulates NF-?B signaling and TNF-induced apoptosis in cultured primary hepatocytes. The question of whether alcohol-induced redox changes inhibit NF-?B dependent gene expression in the presence of TNF in cultured primary hepatocytes will be explored. 2) Determine the effect of chronic alcohol consumption on hepatic redox status, NF-?B redox status, NF-?B signaling pathways, N binding to DNA, and NF-?B dependent gene expression in liver. The relationship between NF-?B signaling and redox status will be explored following chronic alcohol treatment in vivo. A focus of this application is to utilize imaging techniques (e.g., confocal microscopy, flow cytometry) to study redox status and NF-?B signaling simultaneous in the same cell. These methods will allow me to assess possible heterogeneity in redox status and NF-?B signaling that alcohol may induce to the hepatocyte population. The outcome of these studies will provide new insights into the mechanism by which alcohol causes liver damage. Public Health Relevance: The outcome of these studies will provide new insights into the mechanism by which alcohol causes liver damage. The findings of this application may shed light on new therapeutic strategies involving thiol antioxidants for prevention or treatment of alcohol related diseases.

描述（由申请人提供）：我的长期目标是成为一名独立研究者并研究包括酒精性肝病在内的肝病的发病机制。获得 K01 资助将是我成为一名独立研究员的重要一步，因为它将为我提供机会和时间来克服我背景中的许多弱点（缺乏酒精研究的初步数据和出版物、之前缺乏对临床重要模型系统的关注、在细胞信号传导、细胞成像和分子生物学技术方面缺乏经验），从而在不久的将来在酒精研究中成为 R01 的竞争者。通过提供我的工资和研究经费，K01 资助将使我从从事其他 PI 项目的工作中解放出来，让我能够真正专注于酒精，并在细胞信号传导、细胞成像和分子生物学方面变得更有经验。我选择在接下来的一段时间内研究的科学领域是氧化还原生物学、信号转导和使用酒精作为肝损伤模型的肝毒性的界面。 TNF 引起的肝细胞损伤仍然是酒精引起的肝损伤的一个中心点。我们实验室的最新证据表明，通过 GSH 调节或氧化剂处理引起的氧化还原变化通过抑制 NF-κB 转录活性使肝细胞对 TNF 诱导的细胞凋亡敏感。这表明 TNF 分泌与介导肝脏损伤的氧化还原变化之间存在联系。待检验的假设是，酒精引起的氧化还原变化通过抑制 NF-κB 依赖性信号通路，使肝细胞对 TNF 诱导的细胞凋亡敏感。该假设将通过以下具体目标进行检验：1）确定酒精诱导的细胞氧化还原状态的改变如何调节培养的原代肝细胞中的 NF-κB 信号传导和 TNF 诱导的细胞凋亡。将探讨在培养的原代肝细胞中存在 TNF 的情况下酒精诱导的氧化还原变化是否抑制 NF-κB 依赖性基因表达的问题。 2) 确定长期饮酒对肝脏氧化还原状态、NF-κB氧化还原状态、NF-κB信号通路、N与DNA的结合以及肝脏中NF-κB依赖性基因表达的影响。体内慢性酒精治疗后将探讨 NF-κB 信号传导与氧化还原状态之间的关系。该应用的重点是利用成像技术（例如，共聚焦显微镜、流式细胞术）来研究同一细胞中同时的氧化还原状态和 NF-κB 信号传导。这些方法将使我能够评估酒精可能诱导肝细胞群的氧化还原状态和 NF-κB 信号的可能异质性。这些研究的结果将为酒精导致肝损伤的机制提供新的见解。 公共卫生相关性：这些研究的结果将为酒精导致肝损伤的机制提供新的见解。该应用的研究结果可能有助于揭示涉及硫醇抗氧化剂的新治疗策略，用于预防或治疗酒精相关疾病。