Investigation of human apoB48 metabolism using a new, integrated non-steady-state model of apoB48 and apoB100 kinetics

E Björnson, C J Packard, M Adiels, L Andersson, N Matikainen, S Söderlund, J Kahri, C Sihlbom, A Thorsell, H Zhou, M-R Taskinen, J Borén, E Björnson, C J Packard, M Adiels, L Andersson, N Matikainen, S Söderlund, J Kahri, C Sihlbom, A Thorsell, H Zhou, M-R Taskinen, J Borén

Abstract

Background: Triglyceride-rich lipoproteins and their remnants have emerged as major risk factors for cardiovascular disease. New experimental approaches are required that permit simultaneous investigation of the dynamics of chylomicrons (CM) and apoB48 metabolism and of apoB100 in very low-density lipoproteins (VLDL).

Methods: Mass spectrometric techniques were used to determine the masses and tracer enrichments of apoB48 in the CM, VLDL1 and VLDL2 density intervals. An integrated non-steady-state multicompartmental model was constructed to describe the metabolism of apoB48- and apoB100-containing lipoproteins following a fat-rich meal, as well as during prolonged fasting.

Results: The kinetic model described the metabolism of apoB48 in CM, VLDL1 and VLDL2 . It predicted a low level of basal apoB48 secretion and, during fat absorption, an increment in apoB48 release into not only CM but also directly into VLDL1 and VLDL2 . ApoB48 particles with a long residence time were present in VLDL, and in subjects with high plasma triglycerides, these lipoproteins contributed to apoB48 measured during fasting conditions. Basal apoB48 secretion was about 50 mg day-1 , and the increment during absorption was about 230 mg day-1 . The fractional catabolic rates for apoB48 in VLDL1 and VLDL2 were substantially lower than for apoB48 in CM.

Discussion: This novel non-steady-state model integrates the metabolic properties of both apoB100 and apoB48 and the kinetics of triglyceride. The model is physiologically relevant and provides insight not only into apoB48 release in the basal and postabsorptive states but also into the contribution of the intestine to VLDL pool size and kinetics.

Keywords: apolipoprotein B48; kinetics; model; remnants; stable isotope.

Conflict of interest statement

The authors report no duality of interest.

© 2019 The Authors. Journal of Internal Medicine published by John Wiley & Sons Ltd on behalf of Association for Publication of The Journal of Internal Medicine.

Figures

Figure 1
Figure 1
Model fits to experimental data for subject 3 – a representative subject. All tracer‐to‐tracee ratio data (for leucine and glycerol enrichment) in the different fractions are depicted in sub figures a–i, followed by concentration data depicted in subfigures j–s. For model fits to all subjects, see Appendix S1. Plasma leucine enr (enrichment) is plotted in semilogarithmic scale. APE, atom percent excess; TTR, tracer‐to‐tracee ratio.
Figure 2
Figure 2
Model fits to plasma apoB48 and plasma apoB48 enrichment during fasting conditions for the four subjects. Since data were not available in the CM, VLDL 1 and VLDL 2 fraction, the model was fitted to only the total plasma measurements. Total plasma apoB48 concentration is shown in the top row (a–d) for each subject, and total plasma apoB48 enrichment is shown in the bottom row (e–h) for each subject. Modelling of the previous day is indicated with grey background.
Figure 3
Figure 3
The final integrated model structure. Compartments 1‐6 represents apoB100 in VLDL 1 and VLDL 2 fraction; compartments 7–12 represents apoB100‐TG in VLDL 1 and VLDL 2 fraction; compartments 13–19 represents apoB48 in CM, VLDL 1 and VLDL 2 fraction; compartments 20–26 represents apoB48‐TG in CM, VLDL 1 and VLDL 2 fraction. Plasma leucine and plasma glycerol are represented by compartments 27–29 and 38–39, respectively. Blue arrows indicate where postprandial fluxes of apoB48/apoB48‐TG enter the model. The above structure represents a simplified schematic version of the full model (see Figure S2).
Figure 4
Figure 4
(a) Fluxes of apoB48 in CM, VLDL 1 and VLDL 2 and fluxes of apoB100 in VLDL 1 and VLDL 2; (b) Concentrations of apoB48 in CM, VLDL 1 and VLDL 2 and concentration of apoB100 in VLDL 1 and VLDL 2; (c) ApoB48‐TG flux in CM, VLDL 1 and VLDL 2 and apoB100‐TG flux in VLDL 1 and VLDL 2; (d) ApoB48‐TG concentration in CM, VLDL 1 and VLDL 2. Solid lines indicate model predictions and coloured circles indicate experimental data. Modelling of the previous day is indicated with grey background. The concentration and flux (in terms of mass) of apoB100 is higher than that of apoB48 in the VLDL 1/2 fractions. Total apoB48 flux into the CM fraction is higher than the basal apoB48 flux, and postprandial apoB48 flux also constitutes a significant portion of the total postprandial apoB48 flux. The total triglyceride flux into the CM fraction is the biggest source of triglyceride flux. However, VLDL 1‐TG concentration is higher than CM‐TG concentration because of the high CM‐TG FCR.

References

    1. Wulff AB, Nordestgaard BG, Tybjaerg‐Hansen A. APOC3 loss‐of‐function mutations, remnant cholesterol, low‐density lipoprotein cholesterol, and cardiovascular risk: mediation‐ and meta‐analyses of 137 895 individuals. Arterioscler Thromb Vasc Biol 2018; 38: 660–8.
    1. Jorgensen AB, Frikke‐Schmidt R, Nordestgaard BG, Tybjaerg‐Hansen A. Loss‐of‐function mutations in APOC3 and risk of ischemic vascular disease. N Engl J Med 2014; 371: 32–41.
    1. Do R, Willer CJ, Schmidt EM et al Common variants associated with plasma triglycerides and risk for coronary artery disease. Nat Genet 2013; 45: 1345–52.
    1. Morita SY. Metabolism and modification of apolipoprotein b‐containing lipoproteins involved in dyslipidemia and atherosclerosis. Biol Pharm Bull 2016; 39: 1–24.
    1. Adiels M, Packard C, Caslake MJ et al A new combined multicompartmental model for apolipoprotein B‐100 and triglyceride metabolism in VLDL subfractions. J Lipid Res 2005; 46: 58–67.
    1. Adiels M, Taskinen MR, Packard C et al Overproduction of large VLDL particles is driven by increased liver fat content in man. Diabetologia 2006; 49: 755–65.
    1. Schwartz EA, Reaven PD. Lipolysis of triglyceride‐rich lipoproteins, vascular inflammation, and atherosclerosis. Biochim Biophys Acta 2012; 1821: 858–66.
    1. Shojaee‐Moradie F, Ma Y, Lou S, Hovorka R, Umpleby AM. Prandial hypertriglyceridemia in metabolic syndrome is due to an overproduction of both chylomicron and VLDL triacylglycerol. Diabetes 2013; 62: 4063–9.
    1. Chan DC, Wong AT, Pang J, Barrett PH, Watts GF. Inter‐relationships between proprotein convertase subtilisin/kexin type 9, apolipoprotein C‐III and plasma apolipoprotein B‐48 transport in obese subjects: a stable isotope study in the postprandial state. Clin Sci (Lond) 2015; 128: 379–85.
    1. Verges B, Duvillard L, Pais de Barros JP et al Liraglutide reduces postprandial hyperlipidemia by increasing ApoB48 (Apolipoprotein B48) catabolism and by reducing ApoB48 production in patients with type 2 diabetes mellitus. Arterioscler Thromb Vasc Biol 2018; 38: 2198–206.
    1. Welty FK, Lichtenstein AH, Barrett PH, Dolnikowski GG, Schaefer EJ. Human apolipoprotein (Apo) B‐48 and ApoB‐100 kinetics with stable isotopes. Arterioscler Thromb Vasc Biol 1999; 19: 2966–74.
    1. Wong AT, Chan DC, Pang J, Watts GF, Barrett PH. Plasma apolipoprotein B‐48 transport in obese men: a new tracer kinetic study in the postprandial state. J Clin Endocrinol Metab 2014; 99: E122–6.
    1. Wong AT, Chan DC, Barrett PH, Adams LA, Watts GF. Effect of omega‐3 fatty acid ethyl esters on apolipoprotein B‐48 kinetics in obese subjects on a weight‐loss diet: a new tracer kinetic study in the postprandial state. J Clin Endocrinol Metab 2014; 99: E1427–35.
    1. Pang J, Chan DC, Hamilton SJ, Tenneti VS, Watts GF, Barrett PH. Effect of niacin on triglyceride‐rich lipoprotein apolipoprotein B‐48 kinetics in statin‐treated patients with type 2 diabetes. Diabetes Obes Metab 2016; 18: 384–91.
    1. Dash S, Xiao C, Morgantini C, Szeto L, Lewis GF. High‐dose resveratrol treatment for 2 weeks inhibits intestinal and hepatic lipoprotein production in overweight/obese men. Arterioscler Thromb Vasc Biol 2013; 33: 2895–901.
    1. Tremblay AJ, Lamarche B, Hogue JC, Couture P. n‐3 Polyunsaturated fatty acid supplementation has no effect on postprandial triglyceride‐rich lipoprotein kinetics in men with type 2 diabetes. J Diabetes Res 2016; 2016: 2909210.
    1. Padilla N, Maraninchi M, Beliard S et al Effects of bariatric surgery on hepatic and intestinal lipoprotein particle metabolism in obese, nondiabetic humans. Arterioscler Thromb Vasc Biol 2014; 34: 2330–7.
    1. Hogue JC, Lamarche B, Tremblay AJ, Bergeron J, Gagne C, Couture P. Evidence of increased secretion of apolipoprotein B‐48‐containing lipoproteins in subjects with type 2 diabetes. J Lipid Res 2007; 48: 1336–42.
    1. Singh SA, Andraski AB, Pieper B et al Multiple apolipoprotein kinetics measured in human HDL by high‐resolution/accurate mass parallel reaction monitoring. J Lipid Res 2016; 57: 714–28.
    1. Pan Y, Zhou H, Mahsut A et al Static and turnover kinetic measurement of protein biomarkers involved in triglyceride metabolism including apoB48 and apoA5 by LC/MS/MS. J Lipid Res 2014; 55: 1179–87.
    1. Lindgren F, Jensen L, Hatch F. The isolation and quantitative analysis of serum lipoproteins In: Nelson G, editor. Blood Lipids and Lipoproteins: Quantitation, Composition and Metabolism. New York: John Wiley and Sons, 1972; 181–274.
    1. Anderson NL, Anderson NG, Haines LR, Hardie DB, Olafson RW, Pearson TW. Mass spectrometric quantitation of peptides and proteins using Stable Isotope Standards and Capture by Anti‐Peptide Antibodies (SISCAPA). J Proteome Res 2004; 3: 235–44.
    1. Chan DC, Watts GF, Nguyen MN, Barrett PH. Factorial study of the effect of n‐3 fatty acid supplementation and atorvastatin on the kinetics of HDL apolipoproteins A‐I and A‐II in men with abdominal obesity. Am J Clin Nutr 2006; 84: 37–43.
    1. Huttunen JK, Ehnholm C, Kinnunen PK, Nikkila EA. An immunochemical method for the selective measurement of two triglyceride lipases in human postheparin plasma. Clin Chim Acta 1975; 63: 335–47.
    1. Mansbach CM 2nd, Parthasarathy S. A re‐examination of the fate of glyceride‐glycerol in neutral lipid absorption and transport. J Lipid Res 1982; 23: 1009–19.
    1. Thomson ABR, Dietschy JM. Intestinal lipid absorption: major extracellular and intracellular events In Johnson LR, ed. Physiology of the Gastrointestinal Tract. New York: Raven Press, 1981; 1147–220.
    1. McKimmie RL, Easter L, Weinberg RB. Acyl chain length, saturation, and hydrophobicity modulate the efficiency of dietary fatty acid absorption in adult humans. Am J Physiol Gastrointest Liver Physiol 2013; 305: G620–7.
    1. Wang TY, Liu M, Portincasa P, Wang DQ. New insights into the molecular mechanism of intestinal fatty acid absorption. Eur J Clin Invest 2013; 43: 1203–23.
    1. Mu H, Hoy CE. The digestion of dietary triacylglycerols. Prog Lipid Res 2004; 43: 105–33.
    1. Gaudet D, Alexander VJ, Baker BF et al Antisense inhibition of apolipoprotein C‐III in patients with hypertriglyceridemia. N Engl J Med 2015; 373: 438–47.
    1. Dewey FE, Gusarova V, Dunbar RL et al Genetic and pharmacologic inactivation of ANGPTL3 and cardiovascular disease. N Engl J Med 2017; 377: 211–21.
    1. Graham MJ, Lee RG, Brandt TA et al Cardiovascular and metabolic effects of ANGPTL3 antisense oligonucleotides. N Engl J Med 2017; 377: 222–32.
    1. Patterson BW, Mittendorfer B, Elias N, Satyanarayana R, Klein S. Use of stable isotopically labeled tracers to measure very low density lipoprotein‐triglyceride turnover. J Lipid Res 2002; 43: 223–33.
    1. Zech LA, Grundy SM, Steinberg D, Berman M. Kinetic model for production and metabolism of very low density lipoprotein triglycerides. Evidence for a slow production pathway and results for normolipidemic subjects. J Clin Invest 1979; 63: 1262–73.
    1. Barrett PH, Baker N, Nestel PJ. Model development to describe the heterogeneous kinetics of apolipoprotein B and triglyceride in hypertriglyceridemic subjects. J Lipid Res 1991; 32: 743–62.
    1. Malmstrom R, Packard CJ, Caslake M et al Defective regulation of triglyceride metabolism by insulin in the liver in NIDDM. Diabetologia 1997; 40: 454–62.
    1. Welty FK, Lichtenstein AH, Barrett PH, Dolnikowski GG, Schaefer EJ. Interrelationships between human apolipoprotein A‐I and apolipoproteins B‐48 and B‐100 kinetics using stable isotopes. Arterioscler Thromb Vasc Biol 2004; 24: 1703–7.
    1. Sakai N, Uchida Y, Ohashi K et al Measurement of fasting serum apoB‐48 levels in normolipidemic and hyperlipidemic subjects by ELISA. J Lipid Res 2003; 44: 1256–62.
    1. Lemieux S, Fontani R, Uffelman KD, Lewis GF, Steiner G. Apolipoprotein B‐48 and retinyl palmitate are not equivalent markers of postprandial intestinal lipoproteins. J Lipid Res 1998; 39: 1964–71.
    1. Smith D, Watts GF, Dane‐Stewart C, Mamo JC. Post‐prandial chylomicron response may be predicted by a single measurement of plasma apolipoprotein B48 in the fasting state. Eur J Clin Invest 1999; 29: 204–9.
    1. Lorec AM, Juhel C, Pafumi Y et al Determination of apolipoprotein B‐48 in plasma by a competitive ELISA. Clin Chem 2000; 46: 1638–42.
    1. Karpe F, Hamsten A. Determination of apolipoproteins B‐48 and B‐100 in triglyceride‐rich lipoproteins by analytical SDS‐PAGE. J Lipid Res 1994; 35: 1311–7.
    1. Schneeman BO, Kotite L, Todd KM, Havel RJ. Relationships between the responses of triglyceride‐rich lipoproteins in blood plasma containing apolipoproteins B‐48 and B‐100 to a fat‐containing meal in normolipidemic humans. Proc Natl Acad Sci U S A 1993; 90: 2069–73.
    1. Lovegrove JA, Isherwood SG, Jackson KG, Williams CM, Gould BJ. Quantitation of apolipoprotein B‐48 in triacylglycerol‐rich lipoproteins by a specific enzyme‐linked immunosorbent assay. Biochim Biophys Acta 1996; 1301: 221–9.
    1. Smith D, Proctor SD, Mamo JC. A highly sensitive assay for quantitation of apolipoprotein B48 using an antibody to human apolipoprotein B and enhanced chemiluminescence. Ann Clin Biochem 1997; 34: 185–9.
    1. D'Aquila T, Hung YH, Carreiro A, Buhman KK. Recent discoveries on absorption of dietary fat: Presence, synthesis, and metabolism of cytoplasmic lipid droplets within enterocytes. Biochim Biophys Acta 2016; 1861: 730–47.
    1. Guo Q, Avramoglu RK, Adeli K. Intestinal assembly and secretion of highly dense/lipid‐poor apolipoprotein B48‐containing lipoprotein particles in the fasting state: evidence for induction by insulin resistance and exogenous fatty acids. Metabolism 2005; 54: 689–97.
    1. Xiao C, Stahel P, Carreiro AL, Buhman KK, Lewis GF. Recent advances in triacylglycerol mobilization by the gut. Trends Endocrinol Metab 2018; 29: 151–63.
    1. Vance JE, Vance DE. Biochemistry of Lipids, Lipoproteins and Membranes. Amsterdam, Netherlands: Elsevier, 2008.
    1. Kindel T, Lee DM, Tso P. The mechanism of the formation and secretion of chylomicrons. Atherosclerosis Supplements 2010; 11: 11–6.
    1. Mahley R, Bennett B, Morre D, Gray M, Thistlethwaite W, LeQuire V. Lipoproteins associated with the Golgi apparatus isolated from epithelial cells of rat small intestine. Lab Invest 1971; 25: 435–44.
    1. Ockner RK, Manning JA. Fatty acid‐binding protein in small intestine identification, isolation, and evidence for its role in cellular fatty acid transport. J Clin Invest 1974; 54: 326–38.
    1. Zilversmit D. The composition and structure of lymph chylomicrons in dog, rat, and man. J Clin Invest 1965; 44: 1610–22.

Source: PubMed

Sponzoři a spolupracovníci

Zdravotní podmínky

Drogové intervence

3
Předplatit