Acute L-Arginine supplementation does not increase nitric oxide production in healthy subjects

Thiago Silveira Alvares, Carlos Adam Conte-Junior, Joab Trajano Silva, Vânia Margaret Flosi Paschoalin, Thiago Silveira Alvares, Carlos Adam Conte-Junior, Joab Trajano Silva, Vânia Margaret Flosi Paschoalin

Abstract

Dietary supplements containing L-arginine have been marketed with the purpose of increasing vasodilatation, and thus, blood and oxygen supply to the exercising muscle. The present study evaluated the acute effect of L-arginine supplementation on indicators of NO production, nitrite (NO2-) + nitrate (NO3-) (NOx), in healthy subjects. Plasma concentrations of asymmetric dimethylarginine (ADMA) and symmetric dimethylarginine (SDMA) have also been addressed. Seventeen healthy males participated in a randomized, double-blind, placebo-controlled study. Blood samples were drawn from a left antecubital vein at baseline (T0). Afterwards, subjects were randomly submittedto 6 g of oral L-arginine supplementation (as L-arginine hydrochloride) or placebo (as corn starch); afterwards, the subjects remained at rest in supine position and blood samples were drawn again at 30 (T1), 60 (T2), 90 (T3) and 120 minutes (T4) after supplementation. To analyze NO production, NO3- was converted to NO2- by nitrate reductase, followed by the derivatization of NO2- with 2,3-diaminonaphthalene. NOx, ADMA and SDMA were analyzed using a high-performance liquid chromatography system and monitored with a fluorescence detector. Two-way ANOVA with repeated measures showed no significant changes in NOx concentrations on the L-arginine group as compared to placebo group at any of the fivetime points (T0: 17.6 ± 3.9 vs 14.6 ± 2.3 μmol/L; T1: 15.8 ± 2.4 vs 14.3 ± 1.7 μmol/L; T2: 16.8 ± 4.9 vs 13.7 ± 2.7 μmol/L; T3: 16.7 ± 3.9 vs 14.6 ± 2.1 μmol/L; T4: 15.1 ± 2.8 vs 13.5 ± 3.5 μmol/L). Furthermore, plasma levels of ADMA and SDMA were not statistically significant between the L-arginine and placebo groups at T0 (0.43 ± 0.19 vs 0.39 ± 0.15 μmol/L and 1.83 ± 1.13 vs 1.70 ± 0.62 μmol/L), respectively. In conclusion, acute L-arginine supplementation does not increase plasma concentration of NOx in healthy individuals with normal plasma concentrations of ADMA.

Figures

Figure 1
Figure 1
Summary of experimental design.
Figure 2
Figure 2
Plasma NOx concentrations (μmol/L). No significant change was observed between groups at any time point. L-Arg = L-arginine supplemented group; Pla = Placebo group.
Figure 3
Figure 3
Total integrated NOx area under curve during the 120 min post supplementation period. No significant change was observed between groups over the time. L-Arg = L-arginine supplemented group; Pla = Placebo group.
Figure 4
Figure 4
Plasma concentration of L-arginine. The symbols ***(P < 0.001), **(P < 0.01) and *(P < 0.05) denotes significantly different from placebo; a(P < 0.01) denotes significantly different from 0 minutes. L-Arg = L-arginine supplemented group; Pla = Placebo group.

References

    1. Kreider RB, Wilborn CD, Taylor L, Campbell B, Almada AL, Collins R, Cooke M, Earnest CP, Greenwood M, Kalman DS, Kerksick CM, Kleiner SM, Leutholtz B, Lopez H, Lowery LM, Mendel R, Smith A, Spano M, Wildman R, Willoughby DS, Ziegenfuss TN, Antonio J. ISSN Exercise & Sport Nutrition Review: Research & Recommendations. J Int Soc Sports Nutr. 2010;7:7. doi: 10.1186/1550-2783-7-7.
    1. Alvares TS, Meirelles CM, Bhambhani YN, Paschoalin VM, Gomes PS. L-arginine as a Potential Ergogenic Aid in Healthy Subjects. Sports Med. 2011;41(3):233–248. doi: 10.2165/11538590-000000000-00000.
    1. Appleton J. Arginine: Clinical potential of a semi-essential amino. Altern Med Rev. 2002;7(6):512–522.
    1. Bode-Böger SM, Scalera F, Ignarro LJ. The L-arginine paradox: importance of the L-arginine/asymmetrical dimethylarginine ratio. PharmacolTher. 2007;114(3):295–306.
    1. Creager M, Gallagher S, Girerd X, Coleman S, Dzau V, Cooke J. L-Arginine improves endothelium-dependent vasodilation in hypercholesterolemic humans. J Clin Invest. 1992;90(4):1248–1253. doi: 10.1172/JCI115987.
    1. Clarkson P, Adams M, Powe A, Donald A, McCredie R, Robinson J, McCarthy S, Keech A, Celermajer D, Deanfiel J. Oral L-Arginnine improves endothelium-dependent dilation in hypercholesterolemic young adults. J Clin Invest. 1996;97(8):1989–1994. doi: 10.1172/JCI118632.
    1. Pieper G, Siebeneich W, Dondlinger L. Short-term oral administration of L-arginine reverses defective endothelium-dependent relaxation and cGMP generation in diabetes. Eur J Pharmacol. 1996;317(2–3):317–320.
    1. Adams M, McCredie R, Jessup W, Robinson J, Sullivan D, Celermajer D. Oral L-Arginine improves endothelium-dependent dilatation and reduces monocyte adhesion to endothelial cells in young men with coronary artery disease. Atherosclerosis. 1997;129(2):261–269. doi: 10.1016/S0021-9150(96)06044-3.
    1. Lerman A, Burnett J, Higano S, McKinley L, Holmes D. Long-term L-Arginine supplementation improves small-vessel coronary endothelial function in humans. Circulation. 1998;97(21):2123–2128. doi: 10.1161/01.CIR.97.21.2123.
    1. Böger R. Asymmetric Dimethylarginine, an Endogenous Inhibitor of Nitric Oxide Synthase, Explains the “L-Arginine Paradox” and Acts as a Novel Cardiovascular Risk Factor. J Nutr. 2004;134(10):2842S–2847S.
    1. Bailey SJ, Winyard PG, Vanhatalo A, Blackwell JR, DiMenna FJ, Wilkerson DP, Jones AM. Acute L-arginine supplementation reduces the O2 cost of moderate-intensity exercise and enhances high-intensity exercise tolerance. J Appl Physiol. 2010;109(5):1394–1403. doi: 10.1152/japplphysiol.00503.2010.
    1. Stevens BR, Godfrey MD, Kaminski TW, Braith RW. High-intensity dynamic human muscle performance enhanced by a metabolic intervention. Med Sci Sports Exerc. 2000;32(12):2102–2108. doi: 10.1097/00005768-200012000-00021.
    1. Buford BN, Koch AJ. Glycine-arginine-alpha-ketoisocaproic acid improves performance of repeated cycling sprints. Med Sci Sports Exerc. 2004;36(4):583–587. doi: 10.1249/01.MSS.0000122075.14060.C4.
    1. Bode-Böger SM, Muke J, Surdacki A, Brabant G, Böger RH, Frölich JC. Oral L-arginine improves endothelial function in healthy individuals older than 70 years. Vasc Med. 2003;8(2):77–81. doi: 10.1191/1358863x03vm474oa.
    1. Forbes SC, Bell GJ. The acute effects of a low and high dose of oral L-arginine supplementation in young active males at rest. Appl Physiol Nutr Metab. 2011;36(3):405–411. doi: 10.1139/h11-035.
    1. Griesenbeck JS, Steck MD, Huber JC. et al.Development of estimates of dietary nitrates, nitrites, and nitrosamines for use with the short Willet food frequency questionnaire. Nutr J. 2009;6:8–16.
    1. Bode-Böger SM, Böger RH, Galland A, Tsikas D, Frölich JC. L-arginine-induced vasodilation in healthy humans: pharmacokinetic-pharmacodynamic relationship. Br J Clin Pharmacol. 1998;46(5):489–497.
    1. Li H, Meininger CJ, Wu G. Rapid determination of nitrite by reversed-phase high-performance liquid chromatography with fluorescence detection. J Chromatogr B Biomed Sci Appl. 2000;746(2):199–207. doi: 10.1016/S0378-4347(00)00328-5.
    1. Tsikas D. Methods of quantitative analysis of the nitric oxide metabolites nitrite and nitrate in human biological fluids. Free Radic Res. 2005;39(8):797–815. doi: 10.1080/10715760500053651.
    1. Ignarro LJ, Fukuto JM, Griscavage JM, Rogers NE, Byrns RE. Oxidation of nitric oxide in aqueous solution to nitrite but not nitrate: Comparison with enzymatically formed nitric oxide from L-arginine. Proc Natl Acad Sci USA. 1993;90(17):8103–8107. doi: 10.1073/pnas.90.17.8103.
    1. Wu G, Meininger CJ. Analysis of Citrulline, Arginine, and Methylarginines using High-Performance Liquid Chromatography. Methods Enzymol. 2008;440:177–189.
    1. Liu TH, Wu CL, Chiang CW, Lo YW, Tseng HF, Chang CK. No effect of short-term arginine supplementation on nitric oxide production, metabolism and performance in intermittent exercise in athletes. J NutrBiochem. 2009;20(6):462–468.
    1. Koppo K, Taes YE, Pottier A, Boone J, Bouckaert J, Derave W. Dietary arginine supplementation speeds pulmonary VO2 kinetics during cycle exercise. Med Sci Sports Exerc. 2009;41(8):1626–1632. doi: 10.1249/MSS.0b013e31819d81b6.
    1. Tang JE, Lysecki PJ, Manolakos JJ, MacDonald MJ, Tarnopolsky MA, Phillips SM. Bolus arginine supplementation affects neither muscle blood flow nor muscle protein synthesis in young men at rest or after resistance exercise. J Nutr. 2011;141(2):195–200. doi: 10.3945/jn.110.130138.
    1. Hickner RC, Fisher JS, Ehsani AA, Kohrt WM. Role of nitric oxide in skeletal muscle blood flow at rest and during dynamic exercise in humans. Am J Physiol. 1997;273(1 Pt 2):H405–H410.
    1. Blum A, Hathaway L, Mincemoyer R, Schenke WH, Kirby M, Csako G, Waclawiw MA, Panza JA, Cannon RO. Effects of oral L-arginine on endothelium-dependent vasodilation and markers of inflammation in healthy postmenopausal women. J Am CollCardiol. 2000;35(2):271–276. doi: 10.1016/S0735-1097(99)00553-7.
    1. Evans RW, Fernstrom JD, Thompson J, Morris SM, Kuller LH. Biochemical responses of healthy subjects during dietary supplementation with L-arginine. J Nutr Biochem. 2004;15(9):534–539. doi: 10.1016/j.jnutbio.2004.03.005.
    1. Schwedhelm E, Maas R, Freese R, Jung D, Lukacs Z, Jambrecina A, Spickler W, Schulze F, Böger RH. Pharmacokinetic and pharmacodynamic properties of oral L-citrulline and L-arginine: impact on nitric oxide metabolism. Br J Clin Pharmacol. 2008;65(1):51–59. doi: 10.1111/j.1365-2125.2007.02990.x.
    1. Sureda A, Cordova A, Ferrer MD. et al.Effects of L-citrulline oral supplementation on polymorphonuclear neutrophils oxidative burst and nitric oxide production after exercise. Free Radic Res. 2009;43(9):828–835. doi: 10.1080/10715760903071664.
    1. Koifman B, Wollman Y, Bogomolny N, Chernichowsky T, Finkelstein A, Peer G, Scherez J, Blum M, Laniado S, Iaina A, Keren G. Improvement of cardiac performance by intravenous infusion of L-arginine in patients with moderate congestive heart failure. J Am Coll Cardiol. 1995;26(5):1251–1256. doi: 10.1016/0735-1097(95)00318-5.
    1. Böger RH, Bode-Böger SM, Thiele W, Creutzig A, Alexander K, Frölich JC. Restoring vascular nitric oxide formation by L-arginine improves the symptoms of intermittent claudication in patients with peripheral arterial occlusive disease. J Am Coll Cardiol. 1998;32(5):1336–1344. doi: 10.1016/S0735-1097(98)00375-1.
    1. Piatti P, Fragasso G, Monti LD, Setola E, Lucotti P, Fermo I, Paroni R, Galluccio E, Pozza G, Chierchia S, Margonato A. Acute intravenous L-arginine infusion decreases endothelin-1 levels and improves endothelial function in patients with angina pectoris and normal coronary arteriograms: correlation with asymmetric dimethylarginine levels. Circulation. 2003;107(3):429–436. doi: 10.1161/01.CIR.0000046489.24563.79.
    1. Lucotti P, Setola E, Monti LD, Galluccio E, Costa S, Sandoli EP, Fermo I, Rabaiotti G, Gatti R, Piatti P. Beneficial effects of a long-term oral L-arginine treatment added to a hypocaloric diet and exercise training program in obese, insulin-resistant type 2 diabetic patients. Am J Physiol Endocrinol Metab. 2006;291(5):E906–E912. doi: 10.1152/ajpendo.00002.2006.
    1. Böger RH, Bode-Böger SM, Szuba A, Tsao PS, Chan JR, Tangphao O, Blaschke TF, Cooke JP. Asymmetric dimethylarginine (ADMA): a novel risk factor for endothelial dysfunction: its role in hypercholesterolemia. Circulation. 1998;98(18):1842–1847. doi: 10.1161/01.CIR.98.18.1842.
    1. Tsikas D, Böger RH, Sandmann J, Bode-Böger SM, Frölich JC. Endogenous nitric oxide synthase inhibitors are responsible for the L-arginine paradox. FEBS Lett. 2000;478(1–2):1–3.
    1. Hov GG, Sagen E, Bigonah A, Asberg A. Health-associated reference values for arginine, asymmetric dimethylarginine (ADMA) and symmetric dimethylarginine (SDMA) measured with high-performance liquid chromatography. Scand J Clin Lab Invest. 2007;67(8):868–876. doi: 10.1080/00365510701429836.
    1. Teerlink T. Measurement of asymmetric dimethylarginine in plasma: methodological considerations and clinical relevance. Clin Chem Lab Med. 2005;43(10):1130–1138.
    1. Horowitz JD, Heresztyn T. An overview of plasma concentrations of asymmetric dimethylarginine (ADMA) in health and disease and in clinical studies: methodological considerations. J Chromatogr B Analyt Technol Biomed Life Sci. 2007;851(1–2):42–50.

Source: PubMed

Sponsorer och medarbetare

Medicinska tillstånd

Läkemedelsinterventioner

3
Prenumerera