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Journal of Lipid Research, Vol. 44, 2006-2014, October 2003
Copyright © 2003 by American Society for Biochemistry and Molecular Biology

Methods

Altered distribution of plasma PAF-AH between HDLs and other lipoproteins in hyperlipidemia and diabetes mellitus

Takeshi Kujiraoka^*, Tadao Iwasaki^*, Mitsuaki Ishihara^*, Mayumi Ito^*, Makoto Nagano^*, Akito Kawaguchi, Sadao Takahashi, Jun Ishi^**, Masahiro Tsuji, Tohru Egashira^*, Irina P. Stepanova, Norman E. Miller and Hiroaki Hattori^1,*

^* Department of Advanced Medical Technology and Development, BML, Inc., Saitama, Japan
Graduate School of Education, Hokkaido University, Sapporo, Japan
Third Department of Internal Medicine, Fukui Medical University, Fukui, Japan
^** Division of Internal Medicine, Hokkaido Hospital for Social Health Insurance, Sapporo, Japan
Institute of Medical Science, Health Sciences University of Hokkaido, Sapporo, Japan
Molecular Pathology Laboratory, International Center for Genetic Engineering and Biotechnology, Trieste, Italy

¹ To whom correspondence should be addressed. e-mail: hhiro{at}bml.co.jp

Platelet-activating factor acetylhydrolase (PAF-AH) is a phospholipase A₂ associated with lipoproteins that hydrolyzes platelet-activating factor (PAF) and oxidized phospholipids. We have developed an ELISA for PAF-AH that is more sensitive than previous methods, and have quantified HDL-associated and non-HDL-associated PAF-AH in healthy, hyperlipidemic, and diabetic subjects. In healthy subjects, plasma total PAF-AH concentration was positively correlated with PAF-AH activity and with plasma total cholesterol, triacylglycerol, LDL cholesterol and apolipoprotein B (apoB) concentrations (all P < 0.01). HDL-associated PAF-AH concentration was correlated positively with plasma apoA-I and HDL cholesterol. Subjects with hyperlipidemia (n = 73) and diabetes mellitus (n = 87) had higher HDL-associated PAF-AH concentrations than did controls (P < 0.01). Non-HDL-associated PAF-AH concentration was lower in diabetic subjects than in controls (P < 0.01). Both hyperlipidemic and diabetic subjects had lower ratios of PAF-AH to apoB (P < 0.01) and higher ratios of PAF-AH to apoA-I (P < 0.01) than did controls.

Our results show that the distribution of PAF-AH mass between HDLs and LDLs is determined partly by the concentrations of the lipoproteins and partly by the mass of enzyme per lipoprotein particle, which is disturbed in hyperlipidemia and diabetes mellitus.

Abbreviations: CHO, Chinese hamster ovary; MAb, monoclonal antibody; PAF-AH, platelet-activating factor acetylhydrolase

Supplementary key words platelet-activating factor acetylhydrolase • high density lipoprotein • low density lipoprotein • lipoprotein-associated phospholipase A₂ • ELISA • Invader^® assay

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