Zebrafish models of vascular inflammation and atherosclerosis

血管炎症和动脉粥样硬化的斑马鱼模型

基本信息

批准号：
7923970
负责人：
Yury Miller
金额：
$ 41.26万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN DIEGO
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-09-01 至 2011-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7923970
关键词：
Acetylcysteine Address Adult Animal Model Animals Antibodies Antioxidants Apolipoprotein E Apolipoproteins B Arterial Fatty Streak Arteries Atherosclerosis Autopsy Biological Assay Blood Vessels Cathepsins Cell Transplants Cells Cholesterol Data Deposition Development Diagnostic Diet Dietary Fats Disease Dominant-Negative Mutation Endothelial Cells Enzymes Epitopes Fatty acid glycerol esters Fishes Gene Expression Genetic Healed Human Hyperlipidemia Image Immunoassay Inflammation Kinetics Knock-out Label Larva Learning Lesion Life Lipids Lipoproteins Low Density Lipoprotein Receptor Matrix Metalloproteinases Measures Microscopic Modeling Modification Monitor Morphology Mus Myocardial Infarction Optics Oryctolagus cuniculus Pathologic Processes Phospholipase Plasma Play Process Proteins Role Screening procedure Stroke Techniques Testing Time Transgenic Organisms Transplantation Variant Vitamin E Water Zebrafish atherogenesis design enzyme activity feeding healing hypercholesterolemia in vivo insight macrophage mimetics novel oxidation toll-like receptor 4 tool uptake vascular inflammation

项目摘要

The aim of this proposal is to establish zebrafish (Danio rerio) models suitable for in vivo studies of certain processes of atherogenesis. It is widely accepted that hypercholesterolemia leads to lipid accumulation in arteries, which in turn induces vascular inflammation and initiates the development of atherosclerosis, the major cause of heart attacks and stroke in humans. Hence, current experimental animal studies of atherosclerosis use mice and rabbits in which a combination of genetic modification and high-fat diet induce extreme hypercholesterolemia and rapid development of atherosclerosis. An important limitation of using mice and rabbits in these studies is that microscopic examination of atherosclerotic lesions is possible only postmortem. An important advantage of using zebrafish is the optical transparency of its larvae. We propose that using zebrafish for studying certain aspects of atherogenesis will enable monitoring the pathologic processes in arteries in a live animal. Specifically, in Aim 1 we will establish a model of vascular inflammation and atherosclerosis in which feeding zebrafish a high-cholesterol diet causes hypercholesterolemia, lipoprotein oxidation and lipid accumulation in the vascular wall. Using adult zebrafish, we will develop a novel animal model to study in vivo lipoprotein oxidation. The optical transparency of zebrafish larvae will enable monitoring the kinetics of lipid accumulation and macrophage recruitment into the vascular wall. Transgenic zebrafish with fluorescent proteins expressed in endothelial cells and in macrophages will be used in these studies. Developing the hypercholesterolemic larvae model will allow us to set up in vivo functional assays to measure the activity of matrix metalloproteinases, cathepsins and phospholipases in live zebrafish larvae (Aim 2). To demonstrate the utility of this unique model we will address a specific mechanism, critical in the development of atherosclerosis. In Aim 3, we will test the importance of lipoprotein oxidation in vascular lipid accumulation and inflammation. Specifically, antioxidants will be added to the fish diet or dissolved in the fish tank water and their effects on lipid and macrophage deposition in the vasculature of transgenic zebrafish, and specific enzyme activities will be studied. In summary, we propose that establishing zebrafish models of vascular inflammation and atherosclerosis will help significantly enhance mechanistic studies of atherosclerosis as well as the design and screening of new therapies.

该提案的目的是建立适合动脉粥样硬化某些过程的体内研究的斑马鱼（Danio rerio）模型。人们普遍认为，高胆固醇血症会导致动脉中脂质积聚，进而诱发血管炎症并引发动脉粥样硬化，这是人类心脏病发作和中风的主要原因。因此，目前动脉粥样硬化的实验动物研究使用小鼠和兔子，其中基因改造和高脂肪饮食的组合诱导极度高胆固醇血症和动脉粥样硬化的快速发展。在这些研究中使用小鼠和兔子的一个重要限制是动脉粥样硬化病变的显微镜检查只能在死后进行。使用斑马鱼的一个重要优势是其幼虫的光学透明度。我们建议使用斑马鱼研究动脉粥样硬化形成的某些方面将能够监测活体动物动脉的病理过程。具体来说，在目标 1 中，我们将建立血管炎症和动脉粥样硬化模型，其中给斑马鱼喂食高胆固醇饮食会导致高胆固醇血症、脂蛋白氧化和血管壁脂质积累。我们将使用成年斑马鱼开发一种新型动物模型来研究体内脂蛋白氧化。斑马鱼幼虫的光学透明度将能够监测脂质积累和巨噬细胞招募到血管壁的动力学。这些研究将使用在内皮细胞和巨噬细胞中表达荧光蛋白的转基因斑马鱼。开发高胆固醇血症幼虫模型将使我们能够建立体内功能测定，以测量活体斑马鱼幼虫中基质金属蛋白酶、组织蛋白酶和磷脂酶的活性（目标 2）。为了证明这种独特模型的实用性，我们将讨论在动脉粥样硬化的发展中至关重要的特定机制。在目标 3 中，我们将测试脂蛋白氧化在血管脂质积累和炎症中的重要性。具体来说，将抗氧化剂添加到鱼的饮食中或溶解在鱼缸水中，及其对转基因斑马鱼血管系统中脂质和巨噬细胞沉积的影响，并将研究特定的酶活性。总之，我们建议建立血管炎症和动脉粥样硬化斑马鱼模型将有助于显着加强动脉粥样硬化的机制研究以及新疗法的设计和筛选。