In vivo visualization and attenuation of oxidized lipid accumulation in hypercholesterolemic zebrafish
In vivo visualization and attenuation of oxidized lipid accumulation in hypercholesterolemic zebrafish
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DOI：
10.1172/jci57755
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发表时间：
2011-12-01
影响因子：
15.9
通讯作者：
Miller, Yury I.
中科院分区：
文献类型：
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作者：
Fang, Longhou;Green, Simone R.;Miller, Yury I.
LDL的氧化修饰是动脉粥样硬化发展的早期病理事件。氧化事件，如丙二醛（MDA）的形成可能会产生特定的，免疫原性表位。事实上，MDA衍生表位的抗体广泛用于动脉粥样硬化研究，并已被证明可以进行心血管成像。在这项研究中，我们设计了一种转基因斑马鱼与温度诱导表达的EGFP标记的单链人单克隆抗体，IK 17，它结合到MDALDL，并使用光学透明的斑马鱼幼虫成像研究。喂食高胆固醇饮食（HCD）补充红色荧光脂质标记的转基因斑马鱼导致血管脂质积累，定量在活动物使用共聚焦显微镜。在热休克诱导IK 17-EGFP表达后，我们测量了IK 17特异性MDA表位在血管中积累的时间过程。用抗氧化剂或回归饮食治疗导致IK 17与血管病变结合减少。有趣的是，表达IK 17-EGFP的幼虫的匀浆与MDA-LDL结合并抑制MDA-LDL与巨噬细胞的结合。此外，IK 17-EGFP的持续表达有效地防止了HCD诱导的血管壁中的脂质积累，表明抗体本身可能具有治疗效果。因此，我们得出结论，HCD喂养的斑马鱼幼虫与条件表达的EGFP标记的氧化特异性抗体提供了一种有效的方法，测试饮食和/或其他治疗抗氧化剂的策略，最终可能适用于人类。
Oxidative modification of LDL is an early pathological event in the development of atherosclerosis. Oxidation events such as malondialdehyde (MDA) formation may produce specific, immunogenic epitopes. Indeed, antibodies to MDA-derived epitopes are widely used in atherosclerosis research and have been demonstrated to enalb Le cardiovascular imaging. In this study, we engineered a transgenic zebrafish with temperature-inducible expression of an EGFP-labeled single-chain human monoclonal antibody, IK17, which binds to MDALDL, and used optically transparent zebrafish larvae for imaging studies. Feeding a high-cholesterol diet (HCD) supplemented with a red fluorescent lipid marker to the transgenic zebrafish resulted in vascular lipid accumulation, quantified in live animals using confocal microscopy. After heat shock-induced expression of IK17-EGFP, we measured the time course of vascular accumulation of IK17-specific MDA epitopes. Treatment with either an antioxidant or a regression diet resulted in reduced IK17 binding to vascular lesions. Interestingly, homogenates of IK17-EGFP-expressing larvae bound to MDA-LDL and inhibited MDA-LDL binding to macrophages. Moreover, sustained expression of IK17-EGFP effectively prevented HCD-induced Lipid accumulation in the vascular wall, suggesting that the antibody itself may have therapeutic effects. Thus, we conclude that HCD-fed zebrafish larvae with conditional expression of EGFP-labeled oxidation-specific antibodies afford an efficient method of testing dietary and/or other therapeutic antioxidant strategies that may ultimately be applied to humans.