Evaluation of a Novel Glutathione Reductase Inhibitor as a Model to Induce Thiol

描述（由申请人提供）：氧化应激在多种心血管疾病的病理生理中起重要作用，包括心肌梗死后的缺血-再灌注损伤、心力衰竭、糖尿病性心肌病和阿霉素诱导的心脏毒性。小分子和蛋白质硫醇以及与硫醇代谢相关的酶已被证明在保护心脏免受氧化应激过程中起着重要作用。由于硫醇在心血管疾病发展中的重要作用，诱导硫醇氧化应激的研究模型是研究这些疾病状态的病理生理以及药物在疾病治疗中的作用的重要工具。本项目将研究一种新型谷胱甘肽还原酶抑制剂2-乙酰氨基-3-[4-（2-乙酰氨基-2-羧乙基磺酰硫-羰基氨基）苯基硫代氨基磺酰磺酰]丙酸（2-AAPA）作为诱导H9c2大鼠心肌细胞和小鼠模型中硫醇氧化应激的研究工具。拟建项目的具体目的是评估2-AAPA在心肌细胞中产生的硫醇氧化应激程度，将该模型中产生的硫醇氧化应激程度与其他药理学氧化应激模型进行比较，以卡维地洛为例，在药物筛选中证明GR抑制剂可用于产生硫醇氧化应激的概念。并通过确定小鼠心脏中硫醇氧化应激的程度，探索2-AAPA在体内模型中的应用。这个项目的主要目标是建立一个心肌细胞中硫醇氧化应激的模型。该模型可用于进一步研究心血管疾病的病理生理学和硫醇改变对这些疾病发展的影响。该模型还可能用于心血管疾病药物的早期筛选，并在分子水平上评估药物对硫醇氧化应激条件的影响。

DESCRIPTION (provided by applicant): Oxidative stress plays an important role in the pathophysiology of a variety of cardiovascular diseases including ischemia-reperfusion injury following a myocardial infarction, heart failure, diabetic cardiomyopathy, and doxorubicin-induced cardiotoxicity. Small molecule and protein thiols as well as enzymes related to thiol metabolism have been shown to have an important role in the protection of the heart against oxidative stress processes. Because of the important role of thiols in the development of cardiovascular diseases, research models for inducing thiol oxidative stress are important tools for studying the pathophysiology of these disease states as well as the role of medications in disease treatment. In this project, a novel glutathione reductase inhibitor, 2-acetylamino-3-[4-(2-acetylamino-2-carboxyethylsulfanylthio- carbonylamino)phenylthiocarbamoylsulfanyl]propionic acid (2-AAPA), will be evaluated as a research tool to induce thiol oxidative stress in H9c2 rat cardiomyocytes and a mouse model. The specific aims of the proposed project are to evaluate the extent of thiol oxidative stress produced by 2-AAPA in cardiomyocytes, to compare the extent of thiol oxidative stress produced in this model to other pharmacologic oxidative stress models, to establish proof of concept that the GR inhibitor can be used to create thiol oxidative stress in medication screening using carvedilol as an example, and to explore the use of 2-AAPA in an in vivo model by determining the extent of thiol oxidative stress produced in mouse heart. The broad objective of this project is to create a model of thiol oxidative stress in cardiomyocytes. This model could be used in further research examining the pathophysiology of cardiovascular diseases and the impact that thiol alterations have on the development of these conditions. This model could also potentially be used in early screening of medications for cardiovascular conditions and to evaluate the effects of medications in thiol oxidative stress conditions at the molecular level. PUBLIC HEALTH RELEVANCE: Cardiovascular disease is a leading cause of morbidity and mortality in the United States. Several of the major cardiovascular disorders, including injury following a myocardial infarction, heart failure, and atherosclerosis, are associated with oxidative stress. This project seeks to establish a research model of thiol alterations in oxidative stress conditions that could be used to further study the development of cardiovascular disorders and to develop new treatment interventions.