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Journal of Lipid Research, Vol. 48, 1175-1189, May 2007
Copyright © 2007 by American Society for Biochemistry and Molecular Biology

Patient-Oriented Research

Effects of reconstituted HDL on charge-based LDL subfractions as characterized by capillary isotachophoresis

Bo Zhang^1,*, Yoshinari Uehara^*, Satoru Hida^*, Shin-ichiro Miura^*, David L. Rainwater, Masaru Segawa, Koichiro Kumagai^*, Kerry-Anne Rye^**,, and Keijiro Saku^*

^* Department of Cardiology, Fukuoka University School of Medicine, Fukuoka, Japan
Department of Genetics, Southwest Foundation for Biomedical Research, San Antonio, TX
Central Laboratory for Pathology and Morphology, Fukuoka University School of Medicine, Fukuoka, Japan
^** Lipid Research Group, Heart Research Institute, Sydney, New South Wales, Australia
Department of Internal Medicine, University of Sydney, Sydney, New South Wales, Australia
Department of Medicine, University of Melbourne, Melbourne, Victoria, Australia

Published, JLR Papers in Press, February 27, 2007.

¹ To whom correspondence should be addressed. e-mail: bozhang{at}cis.fukuoka-u.ac.jp

ABSTRACT

Modified LDL in human plasma including small, dense LDL (sdLDL) and oxidized LDL carries a more negative charge than unmodified LDL and is atherogenic. We examined the effects of apolipoprotein A-I (apoA-I)/POPC discs on charge-based LDL subfractions as determined by capillary isotachophoresis (cITP). Three normal healthy subjects and seven patients with metabolic disorders were included in the study. LDL in human plasma was separated into two major subfractions, fast- and slow-migrating LDL (fLDL and sLDL), by cITP. Normal LDL was characterized by low fLDL, and mildly oxidized LDL in vitro and mildly modified LDL in human plasma were characterized by increased fLDL. Moderately oxidized LDL in vitro and moderately modified LDL in a patient with hypertriglyceridemia and HDL deficiency were characterized by both increased fLDL and a new LDL subfraction with a faster mobility than fLDL [very-fast-migrating LDL as determined by cITP (vfLDL)]. cITP LDL subfractions with faster electrophoretic mobility (fLDL vs. sLDL, vfLDL vs. fLDL) were associated with an increased content of sdLDL. Incubation of a plasma fraction with d > 1.019 g/ml (depleted of triglyceride-rich lipoproteins) in the presence of apoA-I/POPC discs at 37°C greatly decreased vfLDL and fLDL but increased sLDL. Incubation of whole plasma from patients with an altered distribution of cITP LDL subfractions in the presence of apoA-I/POPC discs also greatly decreased fLDL but increased sLDL. ApoA-I/POPC discs decreased the cITP fLDL level, the free cholesterol concentration, and platelet-activating factor acetylhydrolase activity in the sdLDL subclasses (d = 1.040–1.063 g/ml) and increased the size of LDL. ApoA-I/POPC discs reduced charge-modified LDL in human plasma by remodeling cITP fLDL into sLDL subfractions.

Supplementary key words charge-based low density lipoprotein subfractions • plasma density > 1.019 g/ml fraction • small, dense low density lipoprotein subclasses • low density lipoprotein size

Abbreviations: apoA-I, apolipoprotein A-I; apoB, apolipoprotein B; CETP, cholesteryl ester transfer protein; cITP, capillary isotachophoresis; FC, free cholesterol; fHDL, fast-migrating HDL as determined by cITP; fLDL, fast-migrating LDL as determined by cITP; fTRL, fast-migrating triglyceride-rich lipoprotein as determined by cITP; GGE, gradient gel electrophoresis; HC, hypercholesterolemia; HDLD, HDL deficiency; HDL-PAF-AH, HDL-associated platelet-activating factor acetylhydrolase; HTG, hypertriglyceridemia; idLDL, intermediate density LDL; iHDL, intermediate-migrating HDL as determined by cITP; lbLDL, large, buoyant LDL; LDL(–), electronegative LDL as separated by ion-exchange chromatography; LDL(+), electropositive LDL as separated by ion-exchange chromatography; LDL-PAF-AH, LDL-associated platelet-activating factor acetylhydrolase; NL, normolipidemia; OxLDL Ab, oxidized LDL antibody; PL, phospholipid; sdLDL, small, dense LDL; sHDL, slow-migrating HDL as determined by cITP; sLDL, slow-migrating LDL as determined by cITP; sTRL, slow-migrating triglyceride-rich lipoprotein as determined by cITP; TC, total cholesterol; TG, triglyceride; TRL, triglyceride-rich lipoprotein; vfLDL, very-fast-migrating LDL as determined by cITP; vsTRL, very-slow-migrating triglyceride as determined by cITP

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