Nicotinamide riboside supplementation alters body composition and skeletal muscle acetylcarnitine concentrations in healthy obese humans

Carlijn M E Remie, Kay H M Roumans, Michiel P B Moonen, Niels J Connell, Bas Havekes, Julian Mevenkamp, Lucas Lindeboom, Vera H W de Wit, Tineke van de Weijer, Suzanne A B M Aarts, Esther Lutgens, Bauke V Schomakers, Hyung L Elfrink, Rubén Zapata-Pérez, Riekelt H Houtkooper, Johan Auwerx, Joris Hoeks, Vera B Schrauwen-Hinderling, Esther Phielix, Patrick Schrauwen, Carlijn M E Remie, Kay H M Roumans, Michiel P B Moonen, Niels J Connell, Bas Havekes, Julian Mevenkamp, Lucas Lindeboom, Vera H W de Wit, Tineke van de Weijer, Suzanne A B M Aarts, Esther Lutgens, Bauke V Schomakers, Hyung L Elfrink, Rubén Zapata-Pérez, Riekelt H Houtkooper, Johan Auwerx, Joris Hoeks, Vera B Schrauwen-Hinderling, Esther Phielix, Patrick Schrauwen

Abstract

Background: Nicotinamide riboside (NR) is an NAD+ precursor that boosts cellular NAD+ concentrations. Preclinical studies have shown profound metabolic health effects after NR supplementation.

Objectives: We aimed to investigate the effects of 6 wk NR supplementation on insulin sensitivity, mitochondrial function, and other metabolic health parameters in overweight and obese volunteers.

Methods: A randomized, double-blinded, placebo-controlled, crossover intervention study was conducted in 13 healthy overweight or obese men and women. Participants received 6 wk NR (1000 mg/d) and placebo supplementation, followed by broad metabolic phenotyping, including hyperinsulinemic-euglycemic clamps, magnetic resonance spectroscopy, muscle biopsies, and assessment of ex vivo mitochondrial function and in vivo energy metabolism.

Results: Markers of increased NAD+ synthesis-nicotinic acid adenine dinucleotide and methyl nicotinamide-were elevated in skeletal muscle after NR compared with placebo. NR increased body fat-free mass (62.65% ± 2.49% compared with 61.32% ± 2.58% in NR and placebo, respectively; change: 1.34% ± 0.50%, P = 0.02) and increased sleeping metabolic rate. Interestingly, acetylcarnitine concentrations in skeletal muscle were increased upon NR (4558 ± 749 compared with 3025 ± 316 pmol/mg dry weight in NR and placebo, respectively; change: 1533 ± 683 pmol/mg dry weight, P = 0.04) and the capacity to form acetylcarnitine upon exercise was higher in NR than in placebo (2.99 ± 0.30 compared with 2.40 ± 0.33 mmol/kg wet weight; change: 0.53 ± 0.21 mmol/kg wet weight, P = 0.01). However, no effects of NR were found on insulin sensitivity, mitochondrial function, hepatic and intramyocellular lipid accumulation, cardiac energy status, cardiac ejection fraction, ambulatory blood pressure, plasma markers of inflammation, or energy metabolism.

Conclusions: NR supplementation of 1000 mg/d for 6 wk in healthy overweight or obese men and women increased skeletal muscle NAD+ metabolites, affected skeletal muscle acetylcarnitine metabolism, and induced minor changes in body composition and sleeping metabolic rate. However, no other metabolic health effects were observed.This trial was registered at clinicaltrials.gov as NCT02835664.

Keywords: NAD; acetylcarnitine; body composition; human; insulin sensitivity; metabolic health; mitochondrial function; nicotinamide riboside; obesity.

Copyright © The Author(s) on behalf of the American Society for Nutrition 2020.

Figures

FIGURE 1
FIGURE 1
Study design. In this crossover study, participants were randomly assigned to start with 6 wk NR supplementation or 6 wk placebo treatment. After a washout period of 4–7 wk, participants entered the other intervention arm such that all participants served as their own control. Participants were studied in week 5 (day 32) and in week 6 (day 36–40) of each of the 2 interventions. MRS, magnetic resonance spectroscopy; NR, nicotinamide riboside.
FIGURE 2
FIGURE 2
NAD+ metabolites in skeletal muscle after NR and placebo supplementation. (A) NAD+ concentrations measured in skeletal muscle biopsy specimens by enzymatic assay, n = 8. (B) NAD+ metabolites measured in skeletal muscle biopsy specimens by MS, n = 12. Black bars are NR, open bars are placebo. Data are expressed as mean ± SE. P values are derived from the analysis of the mean within-person changes and the SEM of the within-group changes. **P < 0.01. MeNAM, methylnicotinamide; NAAD, nicotinic acid adenine dinucleotide; NAM, nicotinamide adenosine mononucleotide; NMN, nicotinamide mononucleotide; NR, nicotinamide riboside.
FIGURE 3
FIGURE 3
Skeletal muscle ex vivo mitochondrial respiratory capacity after NR and placebo supplementation. (A) State 2 respiration upon M, MO2, and MG2. (B) ADP-stimulated state 3 respiration upon lipid-derived substrate, MO3. (C) ADP-stimulated state 3 respiration upon CI substrates, MG3. (D) ADP-stimulated state 3 respiration upon parallel electron input to both CI and II, MOG3, MGS3, and MOGS3. (E) State u: maximal FCCP-induced uncoupled respiration. (F) State 4o: oligomycin-induced respiration not coupled to ATP synthesis. (G) Protein content of individual complexes of the electron transport chain. Black bars are NR, open bars are placebo. n = 12. Data are expressed as mean ± SE. C, Complex; FCCP, carbonyl cyanide p-trifluoro-methoxyphenyl hydrazone; M, malate; MGS3, malate + glutamate + succinate; MG2, malate + glutamate; MG3, malate + glutamate; MOG3, malate + octanoyl carnitine + glutamate; MOGS3, malate + octanoyl carnitine + glutamate + succinate; MO2, malate + octanoyl carnitine; MO3, malate + octanoyl carnitine + glutamate; NR, nicotinamide riboside; Oxphos, oxidative phosphorylation.
FIGURE 4
FIGURE 4
Skeletal muscle acylcarnitine concentrations measured in the morning and evening after NR and placebo supplementation. (A) Acetylcarnitine concentrations measured by magnetic resonance spectroscopy in skeletal muscle in the evening during rest, after exercise, and the capacity to form acetylcarnitine expressed as the difference between rest and exercise. (B) C0 and C2 concentrations measured in muscle biopsy specimens taken in the morning during rest. (C) SC, MC, and LC concentrations measured in biopsy specimens taken during rest. Black bars are NR, open bars are placebo. n = 13. Data are expressed as mean ± SE. P values are derived from the analysis of the mean within-person changes and the SEM of the within-group changes. *P < 0.05. C0, free carnitine; C2, acetylcarnitine; LC, sum of long-chain acylcarnitines; MC, sum of medium-chain acylcarnitines; NR, nicotinamide riboside; SC, sum of short-chain acylcarnitines.
FIGURE 5
FIGURE 5
Body composition and SMR after NR and placebo supplementation. (A) Body weight. (B) FM and FFM. (C) SMR. (D) SMR corrected for body weight. (E) SMR corrected for FFM. Black bars are NR, open bars are placebo. Data are expressed as mean ± SE. n = 13. P values are derived from the analysis of the mean within-person changes and the SEM of the within-group changes. *P < 0.05. FFM, fat-free mass; FM, fat mass; NR, nicotinamide riboside; SMR, sleeping metabolic rate.

References

    1. Bieganowski P, Brenner C. Discoveries of nicotinamide riboside as a nutrient and conserved NRK genes establish a Preiss-Handler independent route to NAD+ in fungi and humans. Cell. 2004;117(4):495–502.
    1. Connell NJ, Houtkooper RH, Schrauwen P. NAD+ metabolism as a target for metabolic health: have we found the silver bullet?. Diabetologia. 2019;62(6):888–99.
    1. Houtkooper RH, Auwerx J. Exploring the therapeutic space around NAD+. J Cell Biol. 2012;199(2):205–9.
    1. Imai S, Guarente L. NAD+ and sirtuins in aging and disease. Trends Cell Biol. 2014;24(8):464–71.
    1. Bonkowski MS, Sinclair DA. Slowing ageing by design: the rise of NAD+ and sirtuin-activating compounds. Nat Rev Mol Cell Biol. 2016;17(11):679–90.
    1. Canto C, Auwerx J. Targeting sirtuin 1 to improve metabolism: all you need is NAD+?. Pharmacol Rev. 2012;64(1):166–87.
    1. Fletcher RS, Ratajczak J, Doig CL, Oakey LA, Callingham R, Da Silva Xavier G, Garten A, Elhassan YS, Redpath P, Migaud ME et al. . Nicotinamide riboside kinases display redundancy in mediating nicotinamide mononucleotide and nicotinamide riboside metabolism in skeletal muscle cells. Mol Metab. 2017;6(8):819–32.
    1. Cantó C, Houtkooper RH, Pirinen E, Youn DY, Oosterveer MH, Cen Y, Fernandez-Marcos PJ, Yamamoto H, Andreux PA, Cettour-Rose P et al. . The NAD+ precursor nicotinamide riboside enhances oxidative metabolism and protects against high-fat diet-induced obesity. Cell Metab. 2012;15(6):838–47.
    1. Khan NA, Auranen M, Paetau I, Pirinen E, Euro L, Forsström S, Pasila L, Velagapudi V, Carroll CJ, Auwerx J et al. . Effective treatment of mitochondrial myopathy by nicotinamide riboside, a vitamin B3. EMBO Mol Med. 2014;6(6):721–31.
    1. Trammell SAJ, Weidemann BJ, Chadda A, Yorek MS, Holmes A, Coppey LJ, Obrosov A, Kardon RH, Yorek MA, Brenner C. Nicotinamide riboside opposes type 2 diabetes and neuropathy in mice. Sci Rep. 2016;6:26933.
    1. Shi W, Hegeman MA, van Dartel DAM, Tang J, Suarez M, Swarts H, van der Hee B, Arola L, Keijer J. Effects of a wide range of dietary nicotinamide riboside (NR) concentrations on metabolic flexibility and white adipose tissue (WAT) of mice fed a mildly obesogenic diet. Mol Nutr Food Res. 2017;61(8):1600878.
    1. Frederick DW, Loro E, Liu L, Davila A Jr, Chellappa K, Silverman IM, Quinn WJ 3rd, Gosai SJ, Tichy ED, Davis JG et al. . Loss of NAD homeostasis leads to progressive and reversible degeneration of skeletal muscle. Cell Metab. 2016;24(2):269–82.
    1. Trammell SA, Schmidt MS, Weidemann BJ, Redpath P, Jaksch F, Dellinger RW, Li Z, Abel ED, Migaud ME, Brenner C. Nicotinamide riboside is uniquely and orally bioavailable in mice and humans. Nat Commun. 2016;7:12948.
    1. Yoshino J, Mills KF, Yoon MJ, Imai S. Nicotinamide mononucleotide, a key NAD+ intermediate, treats the pathophysiology of diet- and age-induced diabetes in mice. Cell Metab. 2011;14(4):528–36.
    1. Massudi H, Grant R, Braidy N, Guest J, Farnsworth B, Guillemin GJ. Age-associated changes in oxidative stress and NAD+ metabolism in human tissue. PLoS One. 2012;7(7):e42357.
    1. de Guia RM, Agerholm M, Nielsen TS, Consitt LA, Sogaard D, Helge JW, Larsen S, Brandauer J, Houmard JA, Treebak JT. Aerobic and resistance exercise training reverses age-dependent decline in NAD+ salvage capacity in human skeletal muscle. Physiol Rep. 2019;7(12):e14139.
    1. Dellinger RW, Santos SR, Morris M, Evans M, Alminana D, Guarente L, Marcotulli E. Author correction: repeat dose NRPT (nicotinamide riboside and pterostilbene) increases NAD+ levels in humans safely and sustainably: a randomized, double-blind, placebo-controlled study. NPJ Aging Mech Dis. 2017;4:8.
    1. Airhart SE, Shireman LM, Risler LJ, Anderson GD, Nagana Gowda GA, Raftery D, Tian R, Shen DD, O'Brien KD. An open-label, non-randomized study of the pharmacokinetics of the nutritional supplement nicotinamide riboside (NR) and its effects on blood NAD+ levels in healthy volunteers. PLoS One. 2017;12(12):e0186459.
    1. Martens CR, Denman BA, Mazzo MR, Armstrong ML, Reisdorph N, McQueen MB, Chonchol M, Seals DR. Chronic nicotinamide riboside supplementation is well-tolerated and elevates NAD+ in healthy middle-aged and older adults. Nat Commun. 2018;9(1):1286.
    1. Dollerup OL, Christensen B, Svart M, Schmidt MS, Sulek K, Ringgaard S, Stødkilde-Jørgensen H, Møller N, Brenner C, Treebak JT et al. . A randomized placebo-controlled clinical trial of nicotinamide riboside in obese men: safety, insulin-sensitivity, and lipid-mobilizing effects. Am J Clin Nutr. 2018;108(2):343–53.
    1. Conze D, Brenner C, Kruger CL. Safety and metabolism of long-term administration of NIAGEN (nicotinamide riboside chloride) in a randomized, double-blind, placebo-controlled clinical trial of healthy overweight adults. Sci Rep. 2019;9(1):9772.
    1. Conze DB, Crespo-Barreto J, Kruger CL. Safety assessment of nicotinamide riboside, a form of vitamin B3. Hum Exp Toxicol. 2016;35(11):1149–60.
    1. Dollerup OL, Chubanava S, Agerholm M, Søndergård SD, Altintaş A, Møller AB, Høyer KF, Ringgaard S, Stødkilde-Jørgensen H, Lavery GG et al. . Nicotinamide riboside does not alter mitochondrial respiration, content or morphology in skeletal muscle from obese and insulin-resistant men. J Physiol. 2020;598(4):731–54.
    1. Elhassan YS, Kluckova K, Fletcher RS, Schmidt MS, Garten A, Doig CL, Cartwright DM, Oakey L, Burley CV, Jenkinson N et al. . Nicotinamide riboside augments the aged human skeletal muscle NAD+ metabolome and induces transcriptomic and anti-inflammatory signatures. Cell Rep. 2019;28(7):1717–28..e6.
    1. Baecke JA, Burema J, Frijters JE. A short questionnaire for the measurement of habitual physical activity in epidemiological studies. Am J Clin Nutr. 1982;36(5):936–42.
    1. DeFronzo RA, Tobin JD, Andres R. Glucose clamp technique: a method for quantifying insulin secretion and resistance. Am J Physiol. 1979;237(3):E214–23.
    1. Bergström J, Hermansen L, Hultman E, Saltin B. Diet, muscle glycogen and physical performance. Acta Physiol Scand. 1967;71(2):140–50.
    1. Kato T, Berger SJ, Carter JA, Lowry OH. An enzymatic cycling method for nicotinamide-adenine dinucleotide with malic and alcohol dehydrogenases. Anal Biochem. 1973;53(1):86–97.
    1. van de Weijer T, Phielix E, Bilet L, Williams EG, Ropelle ER, Bierwagen A, Livingstone R, Nowotny P, Sparks LM, Paglialunga S et al. . Evidence for a direct effect of the NAD+ precursor acipimox on muscle mitochondrial function in humans. Diabetes. 2015;64(4):1193–201.
    1. Hoeks J, van Herpen NA, Mensink M, Moonen-Kornips E, van Beurden D, Hesselink MK, Schrauwen P. Prolonged fasting identifies skeletal muscle mitochondrial dysfunction as consequence rather than cause of human insulin resistance. Diabetes. 2010;59(9):2117–25.
    1. Wefers J, van Moorsel D, Hansen J, Connell NJ, Havekes B, Hoeks J, van Marken Lichtenbelt WD, Duez H, Phielix E, Kalsbeek A et al. . Circadian misalignment induces fatty acid metabolism gene profiles and compromises insulin sensitivity in human skeletal muscle. Proc Natl Acad Sci U S A. 2018;115(30):7789–94.
    1. Kuipers H, Verstappen FT, Keizer HA, Geurten P, van Kranenburg G. Variability of aerobic performance in the laboratory and its physiologic correlates. Int J Sports Med. 1985;6(4):197–201.
    1. Lindeboom L, Nabuurs CI, Hesselink MK, Wildberger JE, Schrauwen P, Schrauwen-Hinderling VB. Proton magnetic resonance spectroscopy reveals increased hepatic lipid content after a single high-fat meal with no additional modulation by added protein. Am J Clin Nutr. 2015;101(1):65–71.
    1. Lindeboom L, Bruls YM, van Ewijk PA, Hesselink MK, Wildberger JE, Schrauwen P, Schrauwen-Hinderling VB. Longitudinal relaxation time editing for acetylcarnitine detection with 1H-MRS. Magn Reson Med. 2017;77(2):505–10.
    1. Lindeboom L, Nabuurs CI, Hoeks J, Brouwers B, Phielix E, Kooi ME, Hesselink MK, Wildberger JE, Stevens RD, Koves T et al. . Long-echo time MR spectroscopy for skeletal muscle acetylcarnitine detection. J Clin Invest. 2014;124(11):4915–25.
    1. van de Weijer T, van Ewijk PA, Zandbergen HR, Slenter JM, Kessels AG, Wildberger JE, Hesselink MK, Schrauwen P, Schrauwen-Hinderling VB, Kooi ME. Geometrical models for cardiac MRI in rodents: comparison of quantification of left ventricular volumes and function by various geometrical models with a full-volume MRI data set in rodents. Am J Physiol Heart Circ Physiol. 2012;302(3):H709–15.
    1. Joris PJ, Plat J, Bakker SJ, Mensink RP. Long-term magnesium supplementation improves arterial stiffness in overweight and obese adults: results of a randomized, double-blind, placebo-controlled intervention trial. Am J Clin Nutr. 2016;103(5):1260–6.
    1. Dempster P, Aitkens S. A new air displacement method for the determination of human body composition. Med Sci Sports Exerc. 1995;27(12):1692–7.
    1. Plasqui G, Soenen S, Westerterp-Plantenga MS, Westerterp KR. Measurement of longitudinal changes in body composition during weight loss and maintenance in overweight and obese subjects using air-displacement plethysmography in comparison with the deuterium dilution technique. Int J Obes (Lond). 2011;35(8):1124–30.
    1. Friedewald WT, Levy RI, Fredrickson DS. Estimation of the concentration of low-density lipoprotein cholesterol in plasma, without use of the preparative ultracentrifuge. Clin Chem. 1972;18(6):499–502.
    1. Péronnet F, Massicotte D. Table of nonprotein respiratory quotient: an update. Can J Sport Sci. 1991;16(1):23–9.
    1. Weir JB. New methods for calculating metabolic rate with special reference to protein metabolism. J Physiol. 1949;109(1–2):1–9.
    1. Steele R. Influences of glucose loading and of injected insulin on hepatic glucose output. Ann N Y Acad Sci. 1959;82:420–30.
    1. de Ligt M, Bruls YMH, Hansen J, Habets MF, Havekes B, Nascimento EBM, Moonen-Kornips E, Schaart G, Schrauwen-Hinderling VB, van Marken Lichtenbelt W et al. . Resveratrol improves ex vivo mitochondrial function but does not affect insulin sensitivity or brown adipose tissue in first degree relatives of patients with type 2 diabetes. Mol Metab. 2018;12:39–47.
    1. Timmers S, Konings E, Bilet L, Houtkooper RH, van de Weijer T, Goossens GH, Hoeks J, van der Krieken S, Ryu D, Kersten S et al. . Calorie restriction-like effects of 30 days of resveratrol supplementation on energy metabolism and metabolic profile in obese humans. Cell Metab. 2011;14(5):612–22.
    1. Timmers S, de Ligt M, Phielix E, van de Weijer T, Hansen J, Moonen-Kornips E, Schaart G, Kunz I, Hesselink MK, Schrauwen-Hinderling VB et al. . Resveratrol as add-on therapy in subjects with well-controlled type 2 diabetes: a randomized controlled trial. Diabetes Care. 2016;39:2211–17.
    1. Diguet N, Trammell SAJ, Tannous C, Deloux R, Piquereau J, Mougenot N, Gouge A, Gressette M, Manoury B, Blanc J et al. . Nicotinamide riboside preserves cardiac function in a mouse model of dilated cardiomyopathy. Circulation. 2018;137(21):2256–73.
    1. Verdecchia P, Angeli F, Cavallini C. Ambulatory blood pressure for cardiovascular risk stratification. Circulation. 2007;115(16):2091–3.
    1. Hansen TW, Jeppesen J, Rasmussen S, Ibsen H, Torp-Pedersen C. Ambulatory blood pressure monitoring and risk of cardiovascular disease: a population based study. Am J Hypertens. 2006;19(3):243–50.
    1. Klepochová R, Valkovič L, Gajdošik M, Hochwartner T, Tschan H, Krebs M, Trattnig S, Krššák M. Detection and alterations of acetylcarnitine in human skeletal muscles by 1H MRS at 7 T. Invest Radiol. 2017;52(7):412–18.
    1. Watt MJ, Heigenhauser GJ, Stellingwerff T, Hargreaves M, Spriet LL. Carbohydrate ingestion reduces skeletal muscle acetylcarnitine availability but has no effect on substrate phosphorylation at the onset of exercise in man. J Physiol. 2002;544(3):949–56.
    1. Salic K, Gart E, Seidel F, Verschuren L, Caspers M, van Duyvenvoorde W, Wong KE, Keijer J, Bobeldijk-Pastorova I, Wielinga PY et al. . Combined treatment with l-carnitine and nicotinamide riboside improves hepatic metabolism and attenuates obesity and liver steatosis. Int J Mol Sci. 2019;20(18):4359.

Source: PubMed

스폰서 및 공동 작업자

건강 상태

약물 개입

3
구독하다