INTERMEMBRANE TRANSFER OF CHOLESTEROL HYDROPEROXIDES

氢过氧化物胆固醇的膜间转移

• 批准号: 6639963
• 负责人: Albert Girotti
• 金额: $ 3.96万
• 依托单位: MEDICAL COLLEGE OF WISCONSIN
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-05-05 至 2005-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6639963
• 关键词: cell membrane; cholesterol; cooperative study; cytoprotection; cytotoxicity; detoxification; erythrocyte membrane; free radicals; high performance liquid chromatography; hydrogen peroxide; iron; lipid peroxides; liposomes; membrane transport proteins; oxidative stress; peroxidases; peroxidation; reduction; selenium; thin layer chromatography; tissue /cell culture; vascular endothelium

DESCRIPTION Cholesterol (Ch), the most prominent sterol of mammalian cells, is located predominantly in the plasma membrane, where it comprises 40-45 mol percent of the total lipid. Like other unsaturated lipids, Ch can be oxidized under conditions of cytotoxic oxidative stress. Among the oxidation products, cholesterol hydroperoxides (ChOOHs) are highly important because they can undergo iron-catalyzed 1-electron reduction, thereby triggering chain peroxidation reactions which expand the boundaries of membrane oxidative damage. Alternatively, ChOOHs may undergo glutathione-dependent 2-electron reductive detoxification if a selenoperoxidase (SePX) is encountered. Superimposed on these alternate fates (which are under study in the parent project) is the possibility that ChOOHs can move (or be transferred) from one membrane to another, either within or between cells. Little is known about this prospect. In this proposal to investigate ChOOH transfer, two hypotheses will be tested: (i) ChOOHs, being more polar than Ch, will translocate relatively rapidly from donor to acceptor membranes, and this can be enhanced by a sterol carrier protein; (ii) ChOOH transfer can be hazardous to an acceptor if its antioxidant capabilities are overwhelmed. The specific aims of the project are to examine: (i) spontaneous intermembrane transfer of ChOOHs (ii) sterol carrier protein-mediated intermembrane transfer of ChOOHs, and (iii) spontaneous vs. protein-facilitated transfer to mammalian cells and its cytotoxic consequences. The studies will involve, liposomal membranes, erythrocyte membranes, aortic endothelial cells, selenium and iron manipulation, and two unique analytical techniques developed during past interactions between the Principal Investigator and Co-investigator: high performance liquid chromatography with mercury cathode electrochemical detection, and high performance thin layer chromatography with radioimaging detection. In addition to strengthening collaborative ties between the host and foreign laboratories, these studies will provide insights into previously unrecognized ways by which cells might be exposed to the prooxidant/cytotoxic effects of ChOOHs. The work is biomedically significant because disorders such as atherogenesis and neurodegeneration are associated with lipid peroxidation, and ChOOH transfer may promote this process.

描述 胆固醇（Ch）是哺乳动物细胞中最重要的固醇， 主要存在于质膜中，其中它包含40-45摩尔%的 总脂。像其他不饱和脂质一样，Ch可以在 细胞毒性氧化应激的条件。在氧化产物中， 胆固醇氢过氧化物（ChOOHs）非常重要，因为它们可以 进行铁催化的1-电子还原，从而触发链 过氧化反应扩大了膜氧化的边界， 损害或者，ChOOH可以经历谷胱甘肽依赖性2-电子 如果遇到硒过氧化物酶（SePX），则进行还原性解毒。 叠加在这些交替的命运（这是正在研究中的父 项目）是ChOOH可以从一个 细胞内或细胞之间的另一个膜。对此知之甚少 前景。在这项研究ChOOH转移的提议中，将提出两个假设。 测试：（i）ChOOH，比Ch极性更大，将相对移位 快速从供体膜到受体膜，这可以通过甾醇来增强 载体蛋白;（ii）ChOOH转移可能对受体有害，如果其 抗氧化能力被淹没。该项目的具体目标是 检查：（i）ChOOH的自发膜间转移（ii）甾醇 载体蛋白介导的ChOOH的膜间转移，和（iii） 自发与蛋白质促进的向哺乳动物细胞的转移及其 细胞毒性后果。这些研究将涉及脂质体膜， 红细胞膜，主动脉内皮细胞，硒和铁 操作，以及过去开发的两种独特的分析技术 主要研究者和合作研究者之间的相互作用：高 汞阴极电化学高效液相色谱 检测和放射成像高效薄层色谱法 侦测除了加强东道国与 国外实验室，这些研究将提供深入了解以前 细胞可能暴露于促氧化剂/细胞毒性的未识别的方式 Chooh的影响这项工作具有生物医学意义，因为 由于动脉粥样硬化形成和神经变性与脂质过氧化有关， 而ChOOH的转移可能促进了这一过程。