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(Arteriosclerosis, Thrombosis, and Vascular Biology. 2000;20:2682.)
© 2000 American Heart Association, Inc.

Atherosclerosis and Lipoproteins

Quantification of Cholesteryl Esters in Human and Rabbit Atherosclerotic Plaques by Magic-Angle Spinning ¹³C-NMR

Shaoqing Peng; Wen Guo; Joel D. Morrisett; Michael T. Johnstone; James A. Hamilton

From the Department of Biophysics (S.P., W.G., J.A.H.), Boston University School of Medicine, Boston, Mass; the Departments of Medicine and Biochemistry (J.D.M.), Baylor School of Medicine, Houston, Tex; and the Department of Medicine (M.T.J.), Cardiovascular Division, Beth Israel Medical Center, Boston, Mass.

Correspondence to James A. Hamilton, Department of Biophysics, Boston University School of Medicine, Boston, MA 02118. E-mail Hamilton{at}med-biophd.bu.edu

Abstract—Accumulation of cholesteryl esters (CEs) is a key event in the formation of atherosclerotic plaques. More recent work suggests a role for CEs in plaque rupture leading to thrombosis, which can result in an acute event such as myocardial infarction or stroke. In this study, we present nuclear magnetic resonance (NMR) protocols for quantification of CEs in plaques in situ. Total CEs quantified by ¹³C magic-angle spinning (MAS) NMR in excised plaques from human carotid arteries and rabbit aortic arteries were in good agreement with the amounts determined by subsequent standard chemical assays. The latter analysis is disadvantageous because it requires that plaque lipids be extracted from the tissue, resulting in the loss of all phase information of CEs as well as other major plaque components. With our MAS-NMR protocol, the plaque components are preserved in their native phases. Combining MAS and off-MAS NMR, we were able to quantitatively distinguish isotropic (liquid) CEs from anisotropic (liquid-crystalline) CEs in plaque tissues. In a recent study, we applied a different ¹³C MAS-NMR protocol to quantify crystalline cholesterol monohydrate in plaques. Together, these 2 studies describe a new, noninvasive MAS-NMR strategy for the identification and quantification of the major lipid components in plaques in situ. This approach will be useful for investigation of the relationship between plaque rupture and specific lipids in their biologically relevant phases.

Key Words: NMR • cholesteryl esters • atherosclerotic plaques • plaque rupture • lipid phase

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