LONG-CHAIN ACYL-COA SYNTHETASES IN VASCULAR CELLS

血管细胞中的长链酰基辅酶A合成酶

• 批准号: 6869514
• 负责人: Karin E. Bornfeldt
• 金额: $ 37.9万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-04-01 至 2008-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6869514
• 关键词: RNA interference; acyl coA; atherosclerosis; cholesterol; clinical research; diabetes mellitus; diacylglycerols; disease /disorder model; enzyme activity; enzyme inhibitors; genetically modified animals; growth factor; human subject; laboratory mouse; ligase; lipid metabolism; macrophage; oleate; pathologic process; phosphatidylcholines; phospholipase D; protein isoforms; vascular smooth muscle

DESCRIPTION (provided by applicant): Triglycerides contribute to the increased cardiovascular mortality in people with diabetes. Long-chain fatty acids are released from triglycerides in lesions of atherosclerosis, and are esterified to CoA in order to be utilized by vascular cells through a reaction that is catalyzed by long-chain acyI-CoA synthetases (ACS1-5). Oleic acid (OA), the most common fatty acid in triglycerides, has important pro-atherosclerotic effects in vascular cells. Four questions will be addressed: 1. Which ACS isoforms are expressed in primary smooth muscle cells (SMCs) and macrophages, and are they regulated by glucose and lipids? We hypothesize that primary SMCs and macrophages express several ACS isoforms, and that their expression is differentially regulated by glucose and lipids in vitro and in vivo. For the in vivo studies, we have developed a new transgenic mouse model of diabetes-accelerated atherosclerosis. 2. Is ACS2 necessary for OA incorporation into phosphatidylcholine and generation of OA-enriched 1,2-diacylglycerol following growth factor stimulation in SMCs? OA enhances the mitogenic effects of growth factors in SMCs by generation of OA-enriched 1,2-diacylglycerol (1,2-DAG) following growth factor-stimulation of phospholipase D. We hypothesize that ACS2 mediates OA-incorporation into phosphatidylcholine and generation of OA-enriched 1,2-DAG following growth factor stimulation. We propose to inhibit ACS2 by using ACS inhibitors and RNAi. 3. Is ACS2 necessary for the ability of OA to enhance growth factor-induced proliferation in SMCs? We hypothesize that ACS2 is necessary for the ability of OA to enhance growth factor-induced proliferation in SMCs. Inhibitors of ACS isoforms and RNAi will be used to inhibit OA-mediated potentiation of mitogenic effects of growth factors. 4. Which specific ACS isoform(s) is necessary for OA-induced macrophage death and inhibition of cholesterol efflux? OA induces macrophage death and inhibits cholesterol efflux. We hypothesize that a specific ACS isoform is necessary for these events. ACS inhibitors and RNAi will be used to determine the ACS isoform that mediates OA-induced effects on macrophage death, cholesterol efflux and lipid metabolism. The ACS isoforms in vascular cells have not been studied to date. We expect to identify ACS isoforms required for important biological effects of OA in primary smooth muscle and macrophages.

描述（由申请人提供）：甘油三酯导致糖尿病患者心血管死亡率增加。长链脂肪酸从动脉粥样硬化病变的甘油三酯中释放，并酯化为CoA，以便通过由长链酰基辅酶A合成酶（ACS 1 -5）催化的反应被血管细胞利用。油酸（OA）是甘油三酯中最常见的脂肪酸，在血管细胞中具有重要的促动脉粥样硬化作用。将解决四个问题：1。哪些ACS亚型在原代平滑肌细胞（SMC）和巨噬细胞中表达，它们是否受葡萄糖和脂质的调节？我们假设，主要SMC和巨噬细胞表达几种ACS亚型，并且它们的表达在体外和体内受到葡萄糖和脂质的差异调节。在体内研究方面，我们建立了一种新的糖尿病加速动脉粥样硬化的转基因小鼠模型。2.在SMC中生长因子刺激后，ACS 2对于OA掺入磷脂酰胆碱和生成富含OA的1，2-二酰甘油是必需的吗？OA通过在磷脂酶D的生长因子刺激后产生富含OA的1，2-二酰基甘油（1，2-DAG）来增强SMC中生长因子的促有丝分裂作用。我们假设，ACS 2介导的OA纳入磷脂酰胆碱和生成的OA富集的1，2-DAG生长因子刺激后。我们建议通过使用ACS抑制剂和RNAi来抑制ACS 2。3. ACS 2是OA增强SMC中生长因子诱导的增殖能力所必需的吗？我们假设，ACS 2是必要的OA的能力，以增强生长因子诱导的SMC增殖。ACS同种型和RNAi的抑制剂将用于抑制生长因子的促有丝分裂作用的OA介导的增强。4.哪种特定的ACS亚型是OA诱导的巨噬细胞死亡和胆固醇流出抑制所必需的？OA诱导巨噬细胞死亡并抑制胆固醇流出。我们推测，一个特定的ACS亚型是必要的这些事件。ACS抑制剂和RNAi将用于确定ACS亚型，其介导OA诱导的对巨噬细胞死亡、胆固醇流出和脂质代谢的影响。迄今为止，尚未研究血管细胞中的ACS亚型。我们希望确定ACS亚型所需的重要生物效应OA在原代平滑肌和巨噬细胞。