A High Throughput Biochemical Fluorometric Method for Measuring Lipid Peroxidation in HDL

被引：32
作者：
Kelesidis, Theodoros [1 ]
Roberts, Christian K. [2 ]
Huynh, Diana [1 ]
Martinez-Maza, Otoniel [3 ,5 ]
Currier, Judith S. [1 ]
Reddy, Srinivasa T. [1 ,3 ,4 ]
Yang, Otto O. [1 ,5 ]
机构：
[1] Univ Calif Los Angeles, Dept Med, Los Angeles, CA 90024 USA
[2] Univ Calif Los Angeles, Sch Nursing, Exercise & Metab Dis Res Lab, Translat Sci Sect, Los Angeles, CA 90024 USA
[3] Univ Calif Los Angeles, David Geffen Sch Med, Dept Obstet & Gynecol, Los Angeles, CA 90024 USA
[4] Univ Calif Los Angeles, Dept Mol & Med Pharmacol, Los Angeles, CA 90024 USA
[5] Univ Calif Los Angeles, David Geffen Sch Med, Dept Microbiol Immunol & Mol Genet, Los Angeles, CA 90024 USA
来源：
PLOS ONE | 2014年 / 9卷 / 11期
关键词：
HIGH-DENSITY-LIPOPROTEIN; CHOLESTEROL EFFLUX CAPACITY; APOLIPOPROTEIN-A-I; SUBCLINICAL ATHEROSCLEROSIS; ANTIINFLAMMATORY PROPERTIES; INFLAMMATORY PROPERTIES; OXIDIZED PHOSPHOLIPIDS; ANTIOXIDANT PROPERTIES; METHIONINE SULFOXIDE; REACTIVE OXYGEN;
D O I：
10.1371/journal.pone.0111716
中图分类号：
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号：
07 ; 0710 ; 09 ;
摘要：
Current cell-based assays for determining the functional properties of high-density lipoproteins (HDL) have limitations. We report here the development of a new, robust fluorometric cell-free biochemical assay that measures HDL lipid peroxidation (HDLox) based on the oxidation of the fluorochrome Amplex Red. HDLox correlated with previously validated cell-based (r = 0.47, p<0.001) and cell-free assays (r = 0.46, p<0.001). HDLox distinguished dysfunctional HDL in established animal models of atherosclerosis and Human Immunodeficiency Virus (HIV) patients. Using an immunoaffinity method for capturing HDL, we demonstrate the utility of this novel assay for measuring HDLox in a high throughput format. Furthermore, HDLox correlated significantly with measures of cardiovascular diseases including carotid intima media thickness (r = 0.35, p<0.01) and subendocardial viability ratio (r = -0.21, p = 0.05) and physiological parameters such as metabolic and anthropometric parameters (p<0.05). In conclusion, we report the development of a new fluorometric method that offers a reproducible and rapid means for determining HDL function/quality that is suitable for high throughput implementation.
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页数：17
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