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Journal of Lipid Research, Vol. 48, 2086-2095, September 2007
Copyright © 2007 by American Society for Biochemistry and Molecular Biology

Methods

Development of a novel method to determine very low density lipoprotein kinetics

Iqbal A. R. Al-Shayji^*,, Jason M. R. Gill^1,, Josephine Cooney^*, Samira Siddiqui^*, and Muriel J. Caslake^*

^* Department of Vascular Biochemistry, University of Glasgow, Glasgow, United Kingdom
Institute of Diet, Exercise, and Lifestyle, Institute of Biomedical and Life Sciences, University of Glasgow, Glasgow, United Kingdom
Renal Unit, Glasgow Royal Infirmary, Glasgow, United Kingdom

Published, JLR Papers in Press, June 4, 2007.

¹ To whom correspondence should be addressed. e-mail: j.gill{at}bio.gla.ac.uk

Isotopic tracer methods of determining triglyceride-rich lipoprotein (TRL) kinetics are costly, time-consuming, and labor-intensive. This study aimed to develop a simpler and cost-effective method of obtaining TRL kinetic data, based on the fact that chylomicrons compete with large VLDL (VLDL₁; S_f = 60–400) for the same catalytic pathway. Ten healthy subjects [seven men; fasting triglyceride (TG), 44.3–407.6 mg/dl; body mass index, 21–35 kg/m²] were given an intravenous infusion of a chylomicron-like TG emulsion (Intralipid; 0.1 g/kg bolus followed by 0.1 g/kg/h infusion) for 75–120 min to prevent the clearance of VLDL₁ by lipoprotein lipase. Multiple blood samples were taken during and after infusion for separation of Intralipid, VLDL₁, and VLDL₂ by ultracentrifugation. VLDL₁-apolipoprotein B (apoB) and TG production rates were calculated from their linear increases in the VLDL₁ fraction during the infusion. Intralipid-TG clearance rate was determined from its exponential decay after infusion. The production rates of VLDL₁-apoB and VLDL₁-TG were (mean ± SEM) 25.4 ± 3.9 and 1,076.7 ± 224.7 mg/h, respectively, and the Intralipid-TG clearance rate was 66.9 ± 11.7 pools/day. Kinetic data obtained from this method agree with values obtained from stable isotope methods and show the expected relationships with indices of body fatness and insulin resistance (all P < 0.05). The protocol is relatively quick, inexpensive, and transferable to nonspecialist laboratories.

Supplementary key words Intralipid • triglyceride • apolipoprotein B • production • clearance

Abbreviations: apoB, apolipoprotein B; BMI, body mass index; FCR, fractional catabolic rate; FSR, fractional synthetic rate; HOMA_IR, homeostasis model assessment insulin resistance; S_f, Svedberg flotation rate; TG, triglyceride; TRL, triglyceride-rich lipoprotein

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