HDL stimulates the efflux of peroxidized phospholipids from foam cells and its visualization

HDL 刺激过氧化磷脂从泡沫细胞中流出及其可视化

• 批准号: 13460053
• 负责人: MIYAZAWA Teruo
• 金额: $ 6.27万
• 依托单位: Tohoku University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-13460053/
• 关键词: Atherosclerosis; Hyperlipidemia; Membrane phospholipid; Lipid peroxidation; Peroxidized phospholipid; Oxidized lipoprotein; Macrophage; Oxidized LDL; 高脂血症; 血漿リポタンパク; 過酸化; 蛍光ラベル; 画像化

Atherosclerosis is the process underlying coronary artery disease and cerebrovascular disease, and is a leading cause of morbidity in industrialized countries. With the aim of atherosclerosis prevention, mechanisms of atherosclerosis development as well as its protection by dietary food components have been studied worldwide. Here, we demonstrated for the first time that in patient with hyperlipidemia as a major cause of atherosclerosis, the levels of plasma phosphatidylcholine hydroperoxide (PCOOH; that is a primary oxidation product of phosphatidylcholine which located on the surface of plasma lipoprotein particle) were significantly higher than in the healthy volunteers. This indicated that hyperlipidemic patients possess a substantial amount of minimally-oxidized lipoproteins, and implied that the peroxidation of lipoprotein phospholipids would be especially important for the atherosclerotic lesion formation. In the case for using the human monocyte cell line (THP-1), macrophage-like THP-1 cells effectively phagocytized PCOOH-riched oxidized low density lipoprotein (oxLDL), which resulted in the marked accumulation of PCOOH in the cells. By the addition of high density lipoprotein (HDL) to the oxLDL-treated THP-1 cells, cellular PCOOH efficiently attenuated. Thus, we investigated the incorporation mechanism of PCOOH from oxLDL to HDL, and confirmed the ability of HDL to reduce PCOOH by in vitro cell culture experiments and ultra-sensitive LC-MS analysis. On the other hand, fluorescence-labeled PCOOH was able to prepare experimently, and investigated visualization of such PCOOH (I.e., fluorometrically monitorization of the incorporation of oxLDL into THP-1 cells). The present study confirmed that HDL revealed antioxidative function via the reduction of PCOOH.

动脉粥样硬化是冠状动脉疾病和脑血管疾病的潜在过程，并且是工业化国家发病率的主要原因。以预防动脉粥样硬化为目的，动脉粥样硬化的发展机制以及膳食成分的保护作用在世界范围内得到了研究。在此，我们首次证明了在作为动脉粥样硬化主要原因的高脂血症患者中，血浆磷脂酰胆碱氢过氧化物（PCOH;这是位于血浆脂蛋白颗粒表面的磷脂酰胆碱的主要氧化产物）的水平显著高于健康志愿者。这表明高脂血症患者体内存在大量的微量氧化脂蛋白，提示脂蛋白磷脂的过氧化对动脉粥样硬化病变的形成尤为重要。在使用人单核细胞系（THP-1）的情况下，巨噬细胞样THP-1细胞有效地吞噬富含PCOH的氧化低密度脂蛋白（oxLDL），这导致PCOH在细胞中显著蓄积。通过向oxLDL处理的THP-1细胞中加入高密度脂蛋白（HDL），细胞PCOH有效地减弱。因此，我们研究了PCOH从oxLDL到HDL的掺入机制，并通过体外细胞培养实验和超灵敏LC-MS分析证实了HDL还原PCOH的能力。另一方面，荧光标记的PC 00 H能够实验性地制备，并且研究了这种PC 00 H的可视化（即，荧光测定法监测oxLDL掺入THP-1细胞）。本研究证实，HDL通过还原PCOOH显示抗氧化功能。

项目成果

T.Miyazawa 他13名: "Oxidative stress in the absence of inflammation in a mouse model for hepatitis Cvirus associated hepatocarcinogenesis"Cancer Research. 61・6. 173-179 (2000)

T. Miyazawa 等 13 人：“丙型肝炎病毒相关肝癌发生的小鼠模型中无炎症的氧化应激”，癌症研究 61・6（2000 年）。

Nagashima T, Oikawa S, Hirayama Y, Tokita Y, Sekikawa A, Ishigaki Y, Yamada R, Miyazawa T.: "Increase of serum phosphatidylcholine hydroperoxide dependent on glycemic control in type 2 diabetic patients"Diabetes Research and Clinical Practice. 56(1). 19-2

Nagashima T、Oikawa S、Hirayama Y、Tokita Y、Sekikawa A、Ishigaki Y、Yamada R、Miyazawa T.：“2 型糖尿病患者血清磷脂酰胆碱氢过氧化物的增加依赖于血糖控制”糖尿病研究和临床实践。

T.Miyazawa 他1名: "Enhanced level of n-3 fatty acid in membrane phospholipids induces lipid peroxidation in rats fed dietary docosahexaenoic acid oil"Atherosclerosis. 155・1. 9-18 (2001)

T. Miyazawa 等人：“膜磷脂中 n-3 脂肪酸的水平升高会导致喂食二十二碳六烯酸油的大鼠发生脂质过氧化” 动脉粥样硬化。

宮澤 陽夫: "生体脂質ハイドロパーオキサイドと臨床(特別講座)"日本臨床化学会東北支部会誌. 11・1. 1-4 (2002)

Haruo Miyazawa：“生物脂质氢过氧化物和临床实践（特别讲座）”日本临床化学学会东北分会杂志11/1（2002）。

宮澤陽夫: "生体脂質ハイドロパーオキサイドと臨床(特別講座)"日本臨床化学会東北支部会誌. 11・1. 1-4 (2002)

Haruo Miyazawa：“生物脂质过氧化氢和临床实践（特别讲座）”日本临床化学学会东北分会杂志11/1（2002）。