Effect of Organophoshate Exposure on Cholesteryl Ester Hydrolase

有机磷酸盐暴露对胆固醇酯水解酶的影响

• 批准号: 7908563
• 负责人: MATTHEW K ROSS
• 金额: $ 7.15万
• 依托单位: MISSISSIPPI STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-03 至 2011-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7908563
• 关键词: Active Sites; Aftercare; Age; Agriculture; Agrochemicals; American; Area; Arterial Fatty Streak; Arteries; Atherosclerosis; Bile fluid; Buffers; Carboxylic Ester Hydrolases; Cardiology; Cardiovascular Diseases; Cause of Death; Cell Line; Cell Proliferation; Cell-Free System; Cells; Chemistry; Chlorpyrifos; Cholesterol; Cholesterol Esters; Cholesterol Homeostasis; Congestive Heart Failure; Connective Tissue; Coronary Arteriosclerosis; Coronary artery; Country; Cultured Cells; Data; Databases; Disease; Dose; Environmental Health; Environmental Risk Factor; Enzymes; Excretory function; Exposure to; Family; Florida; Genbank; Genes; Goals; Grant; Heart; High Density Lipoproteins; Human; Hydrolase; Hydrolysis; Hypertension; Incidence; Insecticides; Laboratories; Left; Liver; Low-Density Lipoproteins; Metabolism; Methyl Parathion; Mississippi; Modification; Molecular Target; Myocardial Ischemia; O,O-diethyl O-3,5,6-trichloro-2-pyridyl phosphate; Organophosphates; Outcome; Paraoxon; Parathion; Patients; Pesticides; Pharmacological Treatment; Play; Population; Prevalence; Probability; Process; Proteins; Reaction; Recombinants; Recording of previous events; Regulation; Research; Research Project Grants; Risk; Risk Factors; Role; Serine; Serum; Severities; Smooth Muscle Myocytes; Southeastern United States; Stroke; Structure; Students; System; Testing; Toxic Environmental Substances; Training; Transport Process; United States; United States National Institutes of Health; Universities; Valerates; Ventricular; Work; adduct; base; carboxylesterase; environmental chemical; environmental toxicology; esterase; extracellular; human disease; inhibitor/antagonist; insight; low socioeconomic status; macrophage; member; monocyte; mortality; particle; prevent; response; reverse cholesterol transport; statistics; suicide inhibitor; theories; toxic organophosphate insecticide exposure

DESCRIPTION (provided by applicant): Cardiovascular disease (CVD) in its various forms is the leading cause of death in the United States. Reverse cholesterol transport is a mechanism by which cholesterol present in atherosclerotic plaques within arterial walls is transported to the liver via high density lipoprotein particles for excretion in bile. Recent studies have suggested that human cholesteryl ester hydrolase (CEH), an enzyme that metabolizes cholesteryl esters, plays an important role in the regulation of reverse cholesterol transport. This enzyme is identical to the carboxylesterase CES1. Our long term goal is to understand the role that environmental toxicants such as agricultural chemicals play in human disease. Three commonly used organophosphate (OP) insecticides will be used in this proposed study. The hypothesis to be tested is that exposure to OP insecticides will inhibit the CEH/CES1-catalyzed metabolism of cholesteryl esters, which could therefore increase the risk of developing atherosclerosis. Three aims are proposed: (1) Determine the dose response curve for the oxons of chlorpyrifos, parathion, and methyl parathion that inhibit the cholesterol ester hydrolyzing activity of recombinant CEH/CES1 enzyme; (2) Determine the dose response curve for these same oxons with respect to inhibition of CEH/CES1 activity in a human monocyte/macrophage cell line (THP1); (3) Characterize the CEH/CES1 protein adducts formed after treatment of recombinant CEH/CES1 and THP1 cells with chlorpyrifos oxon. We will determine the potency of the active metabolites (oxons) of three environmentally relevant OP insecticides to inhibit CEH/CES1-catalyzed cholesteryl ester hydrolysis activity in a cell-free system and in cultured cells. Furthermore, we will identify the covalent adduct of the protein that inactivates enzyme function. Several environmental factors may increase the incidence of CVD in humans. The results from this study will provide preliminary insights into whether OP oxon metabolites can directly alter the structure-function of an enzyme involved in cholesterol metabolism, thus leading to an increased probability of a pathological outcome (i.e. atherosclerosis). These studies will determine if active metabolites (oxons) of three environmentally relevant organophosphate insecticides can interfere with cholesterol metabolism.

描述(申请人提供)：各种形式的心血管疾病(CVD)是美国主要的死亡原因。胆固醇反向转运是指动脉壁内动脉粥样硬化斑块中存在的胆固醇通过高密度脂蛋白颗粒转运到肝脏并在胆汁中排泄的一种机制。最近的研究表明，人胆固醇酯水解酶(CEH)是一种代谢胆固醇酯的酶，在调节胆固醇反向转运方面发挥着重要作用。该酶与羧酸酯酶CES1相同。我们的长期目标是了解环境毒物，如农用化学品在人类疾病中所起的作用。三种常用的有机磷(OP)杀虫剂将在这项拟议的研究中使用。需要检验的假设是，暴露于OP杀虫剂将抑制CEH/CES1催化的胆固醇酯代谢，从而可能增加发生动脉粥样硬化的风险。提出了三个目标：(1)确定毒死蜱、对硫磷和甲基对硫磷对重组CEH/CES1酶胆固醇酯水解酶活性的抑制作用的剂量反应曲线；(2)确定这些毒死磷在人单核/巨噬细胞系(THP1)中抑制CEH/CES1活性的剂量反应曲线；(3)表征毒死蜱作用于重组CEH/CES1和THP1细胞后形成的CEH/CES1蛋白加合物。我们将确定三种与环境相关的OP杀虫剂的活性代谢物(氧)在无细胞体系和培养细胞中抑制CEH/CES1催化的胆固醇酯水解活性的效力。此外，我们还将鉴定使酶功能失活的蛋白质的共价加合物。几个环境因素可能会增加人类心血管疾病的发病率。这项研究的结果将提供关于OP Oxon代谢物是否可以直接改变参与胆固醇代谢的酶的结构和功能，从而增加病理结果(即动脉粥样硬化)的可能性的初步见解。这些研究将确定三种与环境相关的有机磷杀虫剂的活性代谢物(氧)是否会干扰胆固醇代谢。