Animal Models for Lipid Metabolism and Atherosclerosis

脂质代谢和动脉粥样硬化的动物模型

• 批准号: 6661835
• 负责人: LAWRENCE CHAN
• 金额: $ 7.5万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 1993
• 资助国家: 美国
• 起止时间: 1993-12-15 至 2005-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6661835
• 关键词: alleles; apolipoprotein E; atherosclerosis; diet; disease /disorder etiology; disease /disorder model; enzyme mechanism; gene expression; gene targeting; genetic manipulation; genetic models; genetic promoter element; genetic techniques; genetically modified animals; immunocytochemistry; laboratory mouse; lipoxygenase; liquid chromatography; low density lipoprotein receptor; macrophage; microinjections; mifepristone; model design /development; nutrition related tag; polymerase chain reaction; transfection

DESCRIPTION (provided by applicant): The primary objective is to advance our knowledge of lipid metabolism and atherosclerosis using animal models. A major area is the role of 12/15-lipoxygenase (LO) in atherosclerosis. 12/l5LOs are lipid-peroxidizing enzymes that oxygenate polyenoic fatty acids to their corresponding hydroperoxy derivatives. 12/15LOs have proatherogenic properties via their capacity to oxidize LDL, as well as antiatherogenic activities via their predominantly anti-inflammatory action. We previously showed that macrophage-specific 15LO overexpression in rabbits protects against atherosclerosis. In pilot experiments we found that, compared to 12/15LO+/+ apoE-/- mice, 12/15LO-/- apoE-/- mice are protected against atherosclerosis, a finding that is at variance with that of. A major goal of this application is to examine the biochemical milieu associated with 12/15LO expression that may underlie the divergent effects of the enzyme on atherosclerosis. Another major area is the role of intracellular lipolysis in lipid homeostasis. We produced perilipin (plin) knockout mice that displayed constitutional activated lipolysis. Perilipin is a major adipose lipid droplet protein that regulates the activity of hormone-sensitive lipase. Plin-/- mice were lean and resistant to diet-induced obesity. Breeding the plin-/- alleles into ob/ob and db/db mice reversed their obesity phenotype. Interestingly, ruptured atherosclerotic plaques in humans and advanced plaques of apoE-/- mice expressed perilipin. We will investigate the role of 12/15LO and perilipin with 5 specific aims. In the first 2 aims, we will study atherosclerosis development in 12/15LO-/- apoE-/- and 12/15LO+/+ apoE-/- mice, and, identify and quantify the eicosanoids and other metabolites in the atherosclerotic aortas from the 2 genotypes. In aims 3 and 4, to obtain mechanistic insight into the obesity-resistant phenotype of plin-/- mice, we will perform biochemical and molecular characterization as well as in vivo stable isotope and energy expenditure experiments on plin+/+ and plin-/- mice with and without ob/ob or db/db background. The creation of non-obese ob/ob and db/db mice (i.e., those that also inherited the plin-/- alleles) provides a unique opportunity to study the role of leptin in lipid and carbohydrate homeostasis in the presence and absence of obesity. Since perilipin, a lipid droplet-associated protein, is expressed in atherosclerotic lesions, in the last aim and in collaboration with we will compare the atherosclerosis susceptibility and plaque morphology of plin-/- and plin+/+ mice bred into apoE-/- and LDLR-/- background. We believe that the mechanistic experiments proposed will advance our understanding of lipid and carbohydrate metabolism, energy homeostasis, and atherosclerosis.

描述（由申请人提供）：主要目标是 利用动物增进我们对脂质代谢和动脉粥样硬化的了解 模型。一个主要领域是 12/15-脂氧合酶 (LO) 在动脉粥样硬化中的作用。 12/l5LO 是脂质过氧化酶，可将多烯脂肪酸氧化成 它们相应的氢过氧衍生物。 12/15LOs 具有致动脉粥样硬化 通过氧化低密度脂蛋白的能力以及抗动脉粥样硬化的能力 通过其主要的抗炎作用发挥活性。我们之前 研究表明，巨噬细胞特异性 15LO 在兔子中的过度表达可以防止 动脉粥样硬化。在试点实验中我们发现，与 12/15LO+/+ 相比 apoE-/- 小鼠、12/15LO-/- apoE-/- 小鼠可预防动脉粥样硬化、 发现与 的不一致。一个主要目标是 此应用程序旨在检查与 12/15LO 相关的生化环境 表达可能是酶不同作用的基础 动脉粥样硬化。另一个主要领域是细胞内脂肪分解的作用 脂质稳态。我们培育出 perilipin (plin) 基因敲除小鼠 体质激活脂肪分解。 Perilipin 是一种主要的脂肪脂滴 调节激素敏感性脂肪酶活性的蛋白质。普林-/-小鼠 身材苗条，对饮食引起的肥胖有抵抗力。培育 plin-/- 等位基因 ob/ob 和 db/db 小鼠逆转了它们的肥胖表型。有趣的是， 动脉粥样硬化斑块破裂 在人类和 apoE-/- 小鼠的晚期斑块中表达 perilipin。我们将 研究 12/15LO 和 perilipin 的作用，有 5 个具体目标。在 前2个目标，我们将研究12/15LO-/- apoE-/-的动脉粥样硬化发展 和 12/15LO+/+ apoE-/- 小鼠，并识别和量化类二十烷酸和其他物质 动脉粥样硬化主动脉中两种基因型的代谢物。在目标 3 和 4、获得对肥胖抵抗表型的机制洞察 plin-/- 小鼠，我们将进行生化和分子表征： 以及 plin+/+ 上的体内稳定同位素和能量消耗实验 以及具有或不具有 ob/ob 或 db/db 背景的 plin-/- 小鼠。的创建 非肥胖 ob/ob 和 db/db 小鼠（即那些也继承了 plin-/- 等位基因）提供了一个独特的机会来研究瘦素在脂质和 在存在和不存在肥胖的情况下碳水化合物稳态。自从 perilipin 是一种脂滴相关蛋白，在动脉粥样硬化中表达 病变，最终目标是与我们合作 比较 plin-/- 和 plin-/- 的动脉粥样硬化易感性和斑块形态 plin+/+ 小鼠在 apoE-/- 和 LDLR-/- 背景下繁殖。我们相信 提出的机械实验将增进我们对脂质和 碳水化合物代谢、能量稳态和动脉粥样硬化。