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Journal of Lipid Research, Vol. 50, 716-722, April 2009
Copyright © 2009 by American Society for Biochemistry and Molecular Biology

HDL is the major lipoprotein carrier of plasma F₂-isoprostanes

Julie M. Proudfoot, Anne E. Barden¹, Wai Mun Loke, Kevin D. Croft, Ian B. Puddey and Trevor A. Mori

School of Medicine and Pharmacology, University of Western Australia, Royal Perth Hospital, Perth WA 6847, Australia

This work was supported by grants from the National Heart Foundation of Australia and the National Health and Medical Research Council of Australia.

Published, JLR Papers in Press, December 2, 2008.

¹ To whom correspondence should be addressed. e-mail: anne.barden{at}uwa.edu.au

Enhanced oxidative stress is implicated in the development of atherosclerosis in humans and animal models. F₂-isoprostanes are formed in vivo via free radical peroxidation of arachidonic acid, and their quantification has allowed assessment of oxidative stress in vivo. F₂-isoprostanes associate with lipids, although their distribution in human plasma lipoproteins is unknown. Our aim was to determine the distribution and levels of F₂-isoprostanes in lipoproteins isolated from human plasma by ultracentrifugation and fast protein liquid chromatography (FPLC). F₂-isoprostanes were significantly higher in HDL compared with LDL or VLDL after isolation by ultracentrifugation or FPLC. Furthermore, HDL₃ particles contained elevated levels of F₂-isoprostanes compared with HDL₂. Platelet activating factor acetylhydrolase (PAF-AH), which hydrolyses esterified F₂-isoprostanes from phospholipids, was predominantly associated with LDL. Reduced F₂-isoprostanes in LDL may be related to higher PAF-AH activity in LDL. Paraoxonase 1 (PON-1) activity was associated with HDL₂ and may be a contributing factor to the lower F₂-isoprostanes in HDL₂ compared with HDL₃. Further studies are required to establish the implications of these findings on HDL function.

Supplementary key words HDL oxidation • oxidatively damaged lipid • PAF-AH • PON-1 • FPLC

Abbreviations: AAPH, 2,2'-azobis(2-methylpropionamidine)dihydrochloride; BHT, butylated hydroxytoluene; FPLC, fast protein liquid chromatography; PAF, platelet activating factor; PAF-AH, platelet activating factor acetylhydrolase; PON-1, paraoxonase 1

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