Cardioprotective Mechanisms of Glutathione S-transferase P

谷胱甘肽 S-转移酶 P 的心脏保护机制

• 批准号: 8053818
• 负责人: Daniel Joseph Conklin
• 金额: $ 37万
• 依托单位: UNIVERSITY OF LOUISVILLE
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-04-11 至 2013-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8053818
• 关键词: Ablation; Accounting; Acrolein; Acute; Affect; Affinity; Aldehydes; Antioxidants; Attenuated; Biochemical Pathway; Cardiac; Cardiac Myocytes; Cardiovascular Pathology; Cardiovascular Physiology; Cardiovascular system; Coronary Occlusions; Coronary heart disease; Doctor of Philosophy; Drug Metabolic Detoxication; Drug resistance; Enzymes; Epidemiology; Excision; Foundations; Future; GSTP1 gene; Gender; Genes; Genetic; Genetic Polymorphism; Glutathione; Glutathione S-Transferase; Glutathione S-Transferase P; Health; Heart; Human; In Situ; Individual; Infarction; Injury; Intervention; Ischemia; Kinetics; Knockout Mice; Lead; Link; Lipid Peroxidation; MAPK8 gene; Measures; Mediating; Metabolic; Metabolic Pathway; Metabolism; Modeling; Mus; Myocardial; Myocardial Infarction; Myocardial Ischemia; Myocardial Reperfusion; Myocardial Reperfusion Injury; Natural regeneration; Oxidative Stress; Pathway interactions; Peroxidases; Peroxides; Phenotype; Population; Post-Translational Protein Processing; Predisposition; Principal Investigator; Production; Protein Isoforms; Proteins; Race; Reactive Oxygen Species; Reduced Glutathione; Relative (related person); Reperfusion Injury; Reperfusion Therapy; Role; Signal Pathway; Signal Transduction; Stress; Sulfenic Acids; Testing; Tissues; Tracer; Transgenic Mice; Transgenic Organisms; Translations; adduct; base; carcinogenesis; cardiovascular disorder risk; detoxication; enzyme substrate; gain of function; heart metabolism; insight; loss of function; novel; overexpression; oxidative damage; oxidized lipid; prevent; programs; protein protein interaction; translational study

DESCRIPTION (provided by applicant): The glutathione S-transferases (GSTs) are a superfamily of ubiquitously expressed enzymes that conjugate reduced glutathione (GSH) with toxic electrophiles. The GST isoforms have also been show to regulate ASK-1 and JNK activation via protein-protein interactions. In human populations, the GST isoforms are distributed in a race- and gender-specific manner. The GSTs are expressed in cardiovascular tissues, but the role of GST in cardiovascular physiology and pathology has not been studied. We propose that GSTP, the most abundant cardiac isoform, is an essential component of myocardial defense against oxidative stress and it protects the heart from acute oxidative damage during ischemia-reperfusion (I/R). To test this hypothesis, we will assess the contribution of GSTP to cardiac protection during I/R in situ (Aim 1), metabolism of lipid peroxidation- derived aldehydes (acrolein and 4-hydroxy-trans-2-nonenal; HNE), which are the preferred endogenous substrates of this enzyme (Aim 2), and aldehyde detoxication (Aim 3). As a loss of function test, GSTP-null mice will be exposed to I/R, while in gain of function tests, transgenic (TG) and TG mice crossed with null mice with cardiac-restricted expression of human polymorphic GSTP1-1 genes will be exposed to I/R and infarction determined. Using tracer kinetics and mass spectrometric analysis, we will measure the extent to which glutathiolation accounts for the metabolism of these aldehydes before, during and after ischemia in the heart (Aim 2). To delineate the contribution of GSTP, we will compare metabolic pathways, including aldehyde conjugation and peroxidase activity, in hearts isolated of GSTP WT, null, and TG mice. In Aim 3, we will determine whether GSTP protects the heart from I/R-induced injury by increasing glutathione conjugation of aldehydes, decreasing aldehyde formation, decreasing aldehyde-protein adducts, decreasing peroxide formation, decreasing oxidative post-translation protein modifications (e.g., sulfenic acid) by glutathiolation, and protecting myocardial GSH level. In addition, we will determine if GSTP regulates JNK activation and JNK signaling thereby investigating a non-catalytic role of GSTP in protein-protein interactions. Successful completion of these studies may lead to the identification of a novel mechanism of cardioprotection by GSTP, which may be useful in assessing the relative ischemic susceptibility of human populations polymorphic in GSTP and in developing more targeted anti-ischemic interventions. PUBLIC HEALTH RELEVANCE The glutathione S-transferases (GSTs) are widely expressed enzymes that detoxify reactive compounds. In human populations, the GSTs are distributed in a race- and gender-specific manner, and GST polymorphisms are associated with increased cardiovascular disease risk. This study will assess if GSTP is an essential component of myocardial defense against injury of heart attacks. Successful completion of these studies may lead to the identification of a novel mechanism of cardioprotection by GSTP, which may be useful in understanding the relative susceptibility of human populations polymorphic in GSTP to coronary heart disease and in developing more targeted cardioprotective interventions.

描述（由申请人提供）：谷胱甘肽S-转移酶（GSTS）是无处不在表达的酶的超家族，它们与有毒的电力相结合，将谷胱甘肽还原（GSH）偶联。还显示了GST同工型通过蛋白质 - 蛋白质相互作用来调节ASK-1和JNK激活。在人类种群中，GST同工型以种族和性别特定的方式分布。 GST在心血管组织中表达，但尚未研究GST在心血管生理和病理学中的作用。我们建议GSTP是最丰富的心脏同工型，它是抗氧化应激的心肌防御的重要组成部分，它可以保护心脏免受缺血 - 再灌注过程中急性氧化损伤（I/R）。 To test this hypothesis, we will assess the contribution of GSTP to cardiac protection during I/R in situ (Aim 1), metabolism of lipid peroxidation- derived aldehydes (acrolein and 4-hydroxy-trans-2-nonenal; HNE), which are the preferred endogenous substrates of this enzyme (Aim 2), and aldehyde detoxication (Aim 3).作为功​​能测试的损失，GSTP-NULL小鼠将暴露于I/R，而在功能测试，转基因（TG）和TG小鼠的获得中，将与无效的小鼠交叉具有人类多态性GSTP1-1基因的心脏限制表达，将暴露于I/R和梗死。使用示踪剂动力学和质谱分析，我们将测量谷胱甘肽在心脏中，期间和之后的这些醛的代谢程度（AIM 2）。为了描述GSTP的贡献，我们将比较与GSTP WT，NULL和TG小鼠分离的心脏中的代谢途径，包括醛结合和过氧化物酶活性。 In Aim 3, we will determine whether GSTP protects the heart from I/R-induced injury by increasing glutathione conjugation of aldehydes, decreasing aldehyde formation, decreasing aldehyde-protein adducts, decreasing peroxide formation, decreasing oxidative post-translation protein modifications (e.g., sulfenic acid) by glutathiolation, and protecting心肌GSH水平。此外，我们将确定GSTP是否调节JNK激活和JNK信号传导，从而研究GSTP在蛋白质 - 蛋白质相互作用中的非催化作用。这些研究的成功完成可能会导致GSTP鉴定出一种新型的心脏保护机制，这对于评估GSTP中人类种群的相对缺血性易感性可能很有用，并在开发了更具靶向的抗缺血性干预措施中很有用。 公共卫生相关性谷胱甘肽S-转移酶（GSTS）是广泛表达的酶，可排毒反应性化合物。在人群中，GST以种族和性别特定的方式分布，GST多态性与心血管疾病风险增加有关。这项研究将评估GSTP是否是针对心脏病损伤的心肌防御的重要组成部分。这些研究的成功完成可能会导致GSTP鉴定出一种新型的心脏保护机制，这可能有助于理解GSTP对冠状动脉疾病的相对敏感性以及开发更具靶向性的心脏保护干预措施的相对敏感性。