Nuclear and cytosolic thioredoxin redox state

核和细胞质硫氧还蛋白氧化还原态

• 批准号: 6509434
• 负责人: WALTER H WATSON
• 金额: $ 4.42万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-03-12 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6509434
• 关键词: NAD(P)H oxidoreductase; antineoplastic antibiotics; cell nucleus; cholecalciferol; cytoplasm; endotoxins; enzyme activity; free radical oxygen; gene induction /repression; glutathione; immunogenetics; immunopharmacology; lipopolysaccharides; monocyte; nuclear factor kappa beta; nutrition related tag; oxidation reduction reaction; oxidative stress; phorbols; thioredoxin; tissue /cell culture; western blottings

DESCRIPTION: (Adapted from the Applicant's Abstract): Reactive oxygen species (ROS) and endotoxin (LPS)-induced cytokine responses have been implicated in the pathogenesis of alcoholic liver disease. A key question in ROS signaling is how an oxidant signal can effect transcriptional activation when the latter is inhibited by oxidants. This can occur if the nuclear redox is controlled independently of the cytoplasmic redox. However, the prevailing literature indicates that the nuclear GSH pool is similar to the cytoplasmic pool because nuclear pores are permeable to small molecular weight compounds. I propose to use thioredoxin (Trx) to study redox in the nucleus. I have established methods to measure the redox of Trx using a novel Western blot technique. My hypothesis is that the redox state of nuclear Trx will differ from cytoplasmic Trx under oxidative conditions induced by physiologic and toxicologic stimuli. I will test this hypothesis by measuring redox of nuclear and cytoplasmic Trx and redox of cellular GSH/GSSG in a human monocyte cell line (THP-1) in response to differentiating agents (Vit D3 and phorbol ester) and LPS. I will determine whether isolated nuclei have the capacity to independently regulate Trx redox state and whether nuclear Trx redox state affects NF-kB-dependent gene expression. The research should provide an important advance in knowledge and also a solid foundation for my future career.

描述：（改编自申请人的摘要）：活性氧（ROS）和内毒素（LPS）诱导的细胞因子反应与酒精性肝病的发病机制有关。 ROS 信号传导的一个关键问题是，当转录激活被氧化剂抑制时，氧化剂信号如何影响转录激活。如果核氧化还原的控制独立于细胞质氧化还原，则可能会发生这种情况。然而，主流文献表明核 GSH 池与细胞质池相似，因为核孔可渗透小分子量化合物。我建议使用硫氧还蛋白（Trx）来研究细胞核中的氧化还原。我已经建立了使用新型蛋白质印迹技术测量 Trx 氧化还原的方法。我的假设是，在生理和毒理学刺激诱导的氧化条件下，核 Trx 的氧化还原状态将不同于细胞质 Trx。我将通过测量人单核细胞系 (THP-1) 中核和细胞质 Trx 的氧化还原以及细胞 GSH/GSSG 的氧化还原对分化剂（维生素 D3 和佛波酯）和 LPS 的反应来检验这一假设。我将确定分离的细胞核是否具有独立调节 Trx 氧化还原状态的能力以及核 Trx 氧化还原状态是​​否影响 NF-kB 依赖性基因表达。这项研究应该为我的知识提供重要的进步，并为我未来的职业生涯奠定坚实的基础。