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摘要
背景:胆固醇流出作为胆固醇逆向转运关键的一个过程现今被看作既是诊断的工具又是治疗动脉粥样硬化具有前景的靶点。体外放射性胆固醇流出试验(CEA)是测定细胞级别流出黄金标准。荧光示踪剂和稳定同位素标记胆固醇在非放射性CEA中作为便捷的工具逐渐得到使用。 结果:我们回顾了CEA试验以及测定巨噬细胞特异性胆固醇流出的胆固醇逆向转运中各种有或无失踪方法的使用。使用稳定同位素标记胆固醇的CEA同样依赖于放射性试验并且特别适用于胆固醇流出和网状胆固醇流出的测定。荧光示踪剂不能完全模拟胆固醇;然而能用于在某种特定条件下的CEA试验。荧光法CEA试验会是高流率的同时能在快速胆固醇池或者带着某种单一细胞溶解性中提供特殊的流出信息。酶催化和层析法CEA试验是网赚胆固醇流出试验,当只用于与特殊受体时可以被应用流出试验。体内检测适合器官级别的胆固醇流出和胆固醇逆向转运的研究。它们包括示踪物巨噬细胞注射,一种现今仅适用于动物模型的方法,并且最近能够测定巨噬细胞胆固醇流出的多室模型整体示踪动力学得到了优化。 结论:尽管我们所理解的胆固醇流出机理的体外试验具有决定性作用,体内试验也高度被认为在全身动脉粥样硬化损伤和胆固醇逆向转运方面研究胆固醇流出有研究潜力。
关键词: 载脂蛋白A-I;胆固醇流出;荧光;高密度脂蛋白;质谱;胆固醇逆向转运
Current Medicinal Chemistry
Title:Cholesterol Efflux and Reverse Cholesterol Transport: Experimental Approaches
Volume: 23 Issue: 34
Author(s): Dmitry Y. Litvinov, Eugeny V. Savushkin, Evdokiya A. Garaeva, Alexander D. Dergunov
Affiliation:
关键词: 载脂蛋白A-I;胆固醇流出;荧光;高密度脂蛋白;质谱;胆固醇逆向转运
摘要: Background: Cholesterol efflux as a key event in reverse cholesterol transport (RCT) is considered now as both diagnostic tool and a promising target for the treatment of atherosclerosis. Radioactive in vitro cholesterol efflux assay (CEA) is the gold standard for determination of efflux at cellular level. Fluorescent tracers and stable isotope-labeled cholesterol gradually come into use as convenient tools for non-radioactive CEAs.
Results: We review the use of various tracer-based and tracer-free methods for CEAs and for measuring RCT with focus on macrophage-specific cholesterol efflux. CEA utilizing stable isotope-labeled cholesterol is equally reliable with radioactive assay and especially well suited for the determination of both cholesterol efflux and net cholesterol flux. Fluorescent tracers cannot fully mimic cholesterol; however, they are successfully applied in CEA in specific well-defined conditions. Fluorescent CEAs can be high throughput and can provide unique information on efflux from fast cholesterol pools or with single cell resolution. Enzymatic and chromatographic CEAs are net cholesterol flux assays, and they can be applied as efflux assays when used with specific acceptors only. In vivo tests are suited for studies of cholesterol efflux and RCT at the level of the organism. They include injection of tracer-loaded macrophages, a method suitable at present for animal models only, and recently invented modification of whole body tracer kinetics with multicompartment modeling that is capable to determine cholesterol efflux from macrophages.
Conclusion: Despite the decisive role of in vitro assays in our understanding of cholesterol efflux mechanism, the in vivo assays are highly desired to study cholesterol efflux in atherosclerotic lesions and RCT in whole body.
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Cite this article as:
Dmitry Y. Litvinov, Eugeny V. Savushkin, Evdokiya A. Garaeva, Alexander D. Dergunov , Cholesterol Efflux and Reverse Cholesterol Transport: Experimental Approaches, Current Medicinal Chemistry 2016; 23 (34) . https://dx.doi.org/10.2174/0929867323666160809093009
DOI https://dx.doi.org/10.2174/0929867323666160809093009 |
Print ISSN 0929-8673 |
Publisher Name Bentham Science Publisher |
Online ISSN 1875-533X |
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