Modification of PDI by 4-HNE and 4-ONE and its Role in Ethanol-Induced ER Stress

4-HNE 和 4-ONE 对 PDI 的修饰及其在乙醇诱导的 ER 应激中的作用

• 批准号: 8130540
• 负责人: James J Galligan
• 金额: $ 2.42万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-09-01 至 2012-07-10
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8130540
• 关键词: 4 hydroxynonenal; Active Sites; Affect; Alcoholic Liver Diseases; Aldehydes; Animal Model; Antioxidants; Biological Assay; Cell Line; Chronic; Cysteine; Data; Detection; Diabetes Mellitus; Disease; Endoplasmic Reticulum; Ensure; Enzymes; Ethanol; Ethanol Metabolism; Generations; Glutathione; Hepatocyte; Human; Immunoprecipitation; In Vitro; Inflammation; Lipid Peroxidation; Liver; Liver diseases; Messenger RNA; Modeling; Modification; Molecular Chaperones; Monitor; Neurodegenerative Disorders; Oxidation-Reduction; Oxidative Stress Induction; Pathogenesis; Phase; Play; Protein Disulfide Isomerase; Proteins; Publishing; Reaction; Reactive Oxygen Species; Regulation; Reporting; Research; Rodent; Rodent Model; Role; Staging; Stress; Survival Rate; Testing; Two-Dimensional Gel Electrophoresis; United States; Work; alcohol response; biological adaptation to stress; chronic alcohol ingestion; covalent bond; design; endoplasmic reticulum stress; feeding; in vivo; interest; non-alcoholic; problem drinker; research study; response; 4 hydroxynonenal; Active Sites; Affect; Alcoholic Liver Diseases; Aldehydes; Animal Model; Antioxidants; Biological Assay; Cell Line; Chronic; Cysteine; Data; Detection; Diabetes Mellitus; Disease; Endoplasmic Reticulum; Ensure; Enzymes; Ethanol; Ethanol Metabolism; Generations; Glutathione; Hepatocyte; Human; Immunoprecipitation; In Vitro; Inflammation; Lipid Peroxidation; Liver; Liver diseases; Messenger RNA; Modeling; Modification; Molecular Chaperones; Monitor; Neurodegenerative Disorders; Oxidation-Reduction; Oxidative Stress Induction; Pathogenesis; Phase; Play; Protein Disulfide Isomerase; Proteins; Publishing; Reaction; Reactive Oxygen Species; Regulation; Reporting; Research; Rodent; Rodent Model; Role; Staging; Stress; Survival Rate; Testing; Two-Dimensional Gel Electrophoresis; United States; Work; alcohol response; biological adaptation to stress; chronic alcohol ingestion; covalent bond; design; endoplasmic reticulum stress; feeding; in vivo; interest; non-alcoholic; problem drinker; research study; response

DESCRIPTION (provided by applicant): The build-up of misfolded and/or unfolded proteins in the endoplasmic reticulum (ER) is characterized as the unfolded protein response (UPR). The pathological consequence of this response is known as endoplasmic reticulum (ER) stress and has recently emerged as a possible mechanism for the initiation and progression of numerous disease states, including the alcoholic liver. The ER-resident molecular chaperone, protein disulfide isomerase (PDI), is a key enzyme in both oxidative folding and isomerization reactions. Published data has shown modification of the active-site of PDI by the reactive aldehyde 4-hydroxynonenal (4-HNE). 4-HNE, as well as 4-oxononenal (4-ONE), are products of lipid peroxidation and have been implicated to have a role in the progression of the alcoholic liver. Through the adduction of active site cysteines, these aldehydes have been shown to cause alterations in the enzymatic activity of numerous proteins. The experiments outlined in this proposal are designed to test the general working hypothesis that covalent adduction of PDI by 4-HNE and 4-ONE increase the erred protein burden in the ER and subsequently induces the ER stress response. The first phase of this proposal is designed to elucidate the role of PDI in the ER stress response in vitro, as well as in a chronic ethanol rodent feeding model. Secondly, the alterations in the redox status of PDI will be examined following chronic ethanol feeding. Finally, alterations in the enzymatic activities of PDI will be examined in vivo following ethanol feeding, as well as in vitro, following treatment with the aforementioned aldehydes. Upon completion, the data provided by this proposal will further elucidate the role of PDI in an ethanol-induced ER stress response. With alcoholic liver disease affecting nearly 2 million people in the United States alone, the mechanisms behind its progression remain far from elucidated. The four-year survival rate for end-stage liver disease remains a staggeringly low 35%, stressing an urgency for unraveling the mechanisms behind its pathogenesis. It is most conceivable that the progression of the alcoholic liver is multi-factorial; however, current research suggests a role for both the ER stress response and lipid peroxidation products.

描述（由申请人提供）：内质网中（ER）中错误折叠和/或展开的蛋白质的堆积为未折叠的蛋白质反应（UPR）。这种反应的病理后果称为内质网（ER）应激，最近已成为包括酒精性肝脏在内的许多疾病状态的启动和进展的可能机制。 ER居住的分子伴侣蛋白二硫化物异构酶（PDI）是氧化折叠和异构化反应中的关键酶。已发表的数据表明，反应性醛4-羟基烯烯（4-HNE）对PDI的活性位点进行了修改。 4-HNE以及4-氧登称（4-ONE）是脂质过氧化产物，并被牵涉到在酒精肝脏的进展中发挥作用。通过加合活动位点半胱氨酸，这些醛已被证明会导致许多蛋白质的酶活性改变。该提案中概述的实验旨在检验一般的工作假设，即4-HNE共价增加PDI，而4-ONE增加了ER中的损坏蛋白质负担，并随后诱导ER应力反应。该提案的第一阶段旨在阐明PDI在体外以及慢性乙醇啮齿动物进食模型中的作用。其次，将在慢性乙醇进食后检查PDI的氧化还原状态的改变。最后，在用上述醛处理后，将在乙醇进食以及体外在体内检查PDI的酶活性的改变。完成后，该提案提供的数据将进一步阐明PDI在乙醇引起的ER应力反应中的作用。仅在美国，酒精性肝病就会影响近200万人，其进展的机制远离阐明。终末期肝病的四年生存率仍然是惊人的35％，这强调了揭示其发病机理背后机制的紧迫性。最想想的是，酒精性肝脏的进展是多因素的。但是，当前的研究表明，ER应力反应和脂质过氧化产物都起作用。