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A novel method for production of lipid hydroperoxide- or oxysterol-rich low-density lipoprotein

Gerry, A. B., Satchell, L. and Leake, D. S. (2008) A novel method for production of lipid hydroperoxide- or oxysterol-rich low-density lipoprotein. Atherosclerosis, 197 (2). pp. 579-587. ISSN 0021-9150

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To link to this article DOI: 10.1016/j.atherosclerosis.2007.08.026

Abstract/Summary

LDL oxidation may be important in atherosclerosis. Extensive oxidation of LDL by copper induces increased uptake by macrophages, but results in decomposition of hydroperoxides, making it more difficult to investigate the effects of hydroperoxides in oxidised LDL on cell function. We describe here a simple method of oxidising LDL by dialysis against copper ions at 4 degrees C, which inhibits the decomposition of hydroperoxides, and allows the production of LDL rich in hydroperoxides (626 +/- 98 nmol/mg LDL protein) but low in oxysterols (3 +/- 1 nmol 7-ketocholesterol/mg LDL protein), whilst allowing sufficient modification (2.6 +/- 0.5 relative electrophoretic mobility) for rapid uptake by macrophages (5.49 +/- 0.75 mu g I-125-labelled hydroperoxide-rich LDL vs. 0.46 +/- 0.04 mu g protein/mg cell protein in 18 h for native LDL). By dialysing under the same conditions, but at 37 degrees C, the hydroperoxides are decomposed extensively and the LDL becomes rich in oxysterols. This novel method of oxidising LDL with high yield to either a hydroperoxide- or oxysterol-rich form by simply altering the temperature of dialysis may provide a useful tool for determining the effects of these different oxidation products on cell function. (C) 2007 Elsevier Ireland Ltd. All rights reserved.

Item Type:Article
Refereed:Yes
Divisions:Faculty of Life Sciences > School of Biological Sciences
Interdisciplinary centres and themes > Institute for Cardiovascular and Metabolic Research (ICMR)
ID Code:9792
Uncontrolled Keywords:atherosclerosis, cholesterol, copper, hydroperoxide, oxysterol, macrophage, oxidised low-density lipoprotein, OXIDATIVE MODIFICATION, OXIDIZED LIPIDS, MACROPHAGES, COPPER, PEROXIDATION, ASCORBATE, RADICALS, PROTEIN, ASSAY, ACID

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