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Dietary supplementation with l-lysine affects body weight and blood hematological and biochemical parameters in rats

  • Chao-Wu Xiao
  • Carla Wood
  • Jesse Bertinato
Original Article
  • 47 Downloads

Abstract

l-Lysine (Lys) is a popular additive in foods, but the physiological effects of excess Lys supplementation are poorly understood and upper limits of safe intake have not been established. The objectives of this study were to examine the effects of dietary supplementation with increasing amounts of Lys on body weight (BW), food intake, and various blood hematological and biochemical parameters in rats. Male Sprague–Dawley rats at 10 weeks of age were assigned to ten diet groups (eight rats/group) and fed diets containing either 7% or 20% casein and supplemented with either 0% (Control), 1.5%, 3%, 6% Lys, or 6% Lys + 3% arginine for 1 week. Rats fed 7% casein with ≥ 1.5% Lys supplementation had lower serum albumin and leptin and higher LDL cholesterol (LDLC), ratios of total cholesterol (TC):HDL cholesterol (HDLC) and LDLC:HDLC than those fed 7% casein Control diet (P < 0.05). Rats fed 7% casein diet supplemented with 3% Lys diet had lower BW gain, food intake, serum alkaline phosphatase activity, and increased mean corpuscular hemoglobin concentration, blood urea nitrogen and serum pancreatic polypeptide compared to rats fed the Control diet (P < 0.05). Addition of 6% Lys in 7% casein caused significant BW loss (P < 0.001) and altered additional parameters. Addition of 6% Lys in a 20% casein diet reduced BW gain and food intake and altered numerous parameters. Arg supplementation normalized many of the endpoints changed by Lys. Collectively, these results show that Lys supplementation affects BW, food intake and a number of hematological and biochemical parameters. These effects of Lys supplementation were confined primarily in diets with lower levels of dietary protein. In the context of a low protein diet (7% casein), levels of Lys supplementation ≥ 1.5% may exert adverse health effects in rats.

Keywords

l-Lysine l-Arginine Supplementation Biomarkers Biochemical parameters Rats 

Abbreviations

AA

Amino acid

Arg

l-Arginine

ALB

Albumin

ALP

Alkaline phosphatase

ALT

Alanine aminotransferase

AMY

Amylase

AST

Aspartate aminotransferase

BUN

Blood urea nitrogen

BW

Body weight

HDLC

High density lipoprotein cholesterol

LDLC

Low density lipoprotein cholesterol

Lys

l-Lysine

MCHC

Mean corpuscular hemoglobin concentration

NO

Nitric oxide

PP

Pancreatic polypeptide

Notes

Acknowledgements

This research was funded by Health Canada. We thank the technicians in the Scientific Service Division (SSD), Food Directorate, Health Canada for their outstanding assistance during the animal experimentation phase. We thank Mr. Dominique Patry (SSD) for measuring the blood hematological and biochemical parameters, and Mr. Philip Griffin for his technical assistance at necropsies.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Informed consent

All authors in this paper have read the final manuscript and approved for publication.

Research involving animals

The animal experimental protocol (ACC#2014-018) was approved by the Health Canada-Ottawa Animal Care Committee, and all animal handling and care followed the guidelines of the Canadian Council for Animal Care.

References

  1. 1.
    Le DT, Chu HD, Le NQ (2016) Improving nutritional quality of plant proteins through genetic engineering. Curr Genom 17:220–229CrossRefGoogle Scholar
  2. 2.
    Hussain T, Abbas S, Khan MA, Scrimshaw NS (2004) Lysine fortification of wheat flour improves selected indices of the nutritional status of predominantly cereal-eating families in Pakistan. Food Nutr Bull 25:114–122CrossRefGoogle Scholar
  3. 3.
    Griffith RS, Norins AL, Kagan C (1978) A multicentered study of lysine therapy in Herpes simplex infection. Dermatologica 156:257–267CrossRefGoogle Scholar
  4. 4.
    Griffith RS, Walsh DE, Myrmel KH, Thompson RW, Behforooz A (1987) Success of L-lysine therapy in frequently recurrent herpes simplex infection. Treat Prophyl Dermatol 175:183–190Google Scholar
  5. 5.
    Mahmood ZA (2015) Microbial amino acids production. In: Harzevili FD, Chen H (eds) Microbial biotechnology: progress and trends. Taylor & Francis Group, Boca Raton, pp 194–199Google Scholar
  6. 6.
    Roomi MW, Ivanov V, Kalinovsky T, Niedzwiecki A, Rath M (2006) Anticancer effect of lysine, proline, arginine, ascorbic acid and green tea extract on human renal adenocarcinoma line 786-0. Oncol Rep 16:943–947PubMedGoogle Scholar
  7. 7.
    Unni US, Raj T, Sambashivaiah S, Kuriyan R, Uthappa S, Vaz M, Regan MM, Kurpad AV (2012) The effect of a controlled 8-week metabolic ward based lysine supplementation on muscle function, insulin sensitivity and leucine kinetics in young men. Clin Nutr 31:903–910CrossRefGoogle Scholar
  8. 8.
    Longo N, Frigeni M, Pasquali M (2016) Carnitine transport and fatty acid oxidation. Biochim Biophys Acta 1863:2422–2435CrossRefGoogle Scholar
  9. 9.
    Jafarnejad A, Bathaie SZ, Nakhjavani M, Hassan MZ, Banasadegh S (2008) The improvement effect of L-Lys as a chemical chaperone on STZ-induced diabetic rats, protein structure and function. Diabetes Metab Res Rev 24:64–73CrossRefGoogle Scholar
  10. 10.
    Fischer M, Hirche F, Kluge H, Eder K (2009) A moderate excess of dietary lysine lowers plasma and tissue carnitine concentrations in pigs. Br J Nutr 101:190–196CrossRefGoogle Scholar
  11. 11.
    Civitelli R, Villareal DT, Agnusdei D, Nardi P, Avioli LV, Gennari C (1992) Dietary L-lysine and calcium metabolism in humans. Nutrition 8:400–405PubMedGoogle Scholar
  12. 12.
    Furst P (1993) Dietary L-lysine supplementation: a promising nutritional tool in the prophylaxis and treatment of osteoporosis. Nutrition 9:71–72PubMedGoogle Scholar
  13. 13.
    Bertinato J, Lavergne C, Vu NA, Plouffe LJ, Wood C, Griffin P, Xiao CW (2016) L-Lysine supplementation does not affect the bioavailability of copper or iron in rats. J Trace Elem Med Biol 38:194–200CrossRefGoogle Scholar
  14. 14.
    Fernandez J, Lopez AB, Wang C, Mishra R, Zhou L, Yaman I, Snider MD, Hatzoglou M (2003) Transcriptional control of the arginine/lysine transporter, cat-1, by physiological stress. J Biol Chem 278:50000–50009CrossRefGoogle Scholar
  15. 15.
    Moncada S, Higgs A (1993) The L-arginine-nitric oxide pathway. N Engl J Med 329:2002–2012CrossRefGoogle Scholar
  16. 16.
    Rosselli M, Keller PJ, Dubey RK (1998) Role of nitric oxide in the biology, physiology and pathophysiology of reproduction. Hum Reprod Update 4:3–24CrossRefGoogle Scholar
  17. 17.
    Cittadini D, Pietropaolo C, Decristofaro D, D’Ayjello CM (1964) In vivo effect of L-lysine on rat liver arginase. Nature 203:643–644CrossRefGoogle Scholar
  18. 18.
    Hevia P, Visek WJ (1980) Liver and serum lipids and lipoproteins of rats fed 5% L-lysine. Lipids 15:95–99CrossRefGoogle Scholar
  19. 19.
    Hevia P, Kari FW, Ulman EA, Visek WJ (1980) Serum and liver lipids in growing rats fed casein with L-lysine. J Nutr 110:1224–1230CrossRefGoogle Scholar
  20. 20.
    Butorov EV (2015) Influence of L-lysine amino acid on the HIV-1 RNA replication in vitro. Antivir Chem Chemother 24:39–46CrossRefGoogle Scholar
  21. 21.
    Zyzak DV, Sanders RA, Stojanovic M, Tallmadge DH, Eberhart BL, Ewald DK, Gruber DC, Morsch TR, Strothers MA, Rizzi GP, Villagran MD (2003) Acrylamide formation mechanism in heated foods. J Agric Food Chem 51:4782–4787CrossRefGoogle Scholar
  22. 22.
    Robert F, Vuataz G, Pollien P, Saucy F, Alonso MI, Bauwens I, Blank I (2004) Acrylamide formation from asparagine under low-moisture Maillard reaction conditions. 1. Physical and chemical aspects in crystalline model systems. J Agric Food Chem 52:6837–6842CrossRefGoogle Scholar
  23. 23.
    Yu M, Ou S, Deng L, H C, Zhang J (2013) Effect of ten amino acids on elimination of acrylamide in a model reaction system. Afri J Food Sci 7:329–333CrossRefGoogle Scholar
  24. 24.
    Kobayashi A, Gomikawa S, amazaki A, ato S, onishi T (2014) Elimination of acrylamide by moderate heat treatment below 120 °C with lysine and cysteine. Food Sci Technol Res 20:979–985CrossRefGoogle Scholar
  25. 25.
    Claeys WL, De VK, Hendrickx ME (2005) Effect of amino acids on acrylamide formation and elimination kinetics. Biotechnol Prog 21:1525–1530CrossRefGoogle Scholar
  26. 26.
    Wang B, Ishihara M, Egashira Y, Ohta T, Sanada H (1999) Effects of various kinds of dietary amino acids on the hepatotoxic action of D-galactosamine in rats. Biosci Biotechnol Biochem 63:319–322CrossRefGoogle Scholar
  27. 27.
    Fico ME, Hassan AS, Milner JA (1982) The influence of excess lysine on urea cycle operation and pyrimidine biosynthesis. J Nutr 112:1854–1861CrossRefGoogle Scholar
  28. 28.
    Lin HY, Chen CC, Chen YJ, Lin YY, Mersmann HJ, Ding ST (2014) Enhanced amelioration of high-fat diet-induced fatty liver by docosahexaenoic acid and lysine supplementations. Biomed Res Int 2014:310981PubMedPubMedCentralGoogle Scholar
  29. 29.
    Lewis AJ, Peo ER, Moser BD, Crenshaw TD (1980) Lysine requirement of pigs weighing 5 to 15 kg fed practical diets with and without added Fat. J Anim Sci 51:361–366CrossRefGoogle Scholar
  30. 30.
    Aherne FX, Nielsen HE (1983) Lysine requirement of pigs weighing 7 to 19 kg liveweight. Can J Anim Sci 63:221–224CrossRefGoogle Scholar
  31. 31.
    Edmonds MS, Baker DH (1987) Failure of excess dietary lysine to antagonize arginine in young pigs. J Nutr 117:1396–1401CrossRefGoogle Scholar
  32. 32.
    Yin J, Han H, Li Y, Liu Z, Zhao Y, Fang R, Huang X, Zheng J, Ren W, Wu F, Liu G, Wu X, Wang K, Sun L, Li C, Li T, Yin Y (2017) Lysine restriction affects feed intake and amino acid metabolism via gut microbiome in piglets. Cell Physiol Biochem 44:1749–1761CrossRefGoogle Scholar
  33. 33.
    Yin J, Li Y, Han H, Liu Z, Zeng X, Li T, Yin Y (2018) Long-term effects of lysine concentration on growth performance, intestinal microbiome, and metabolic profiles in a pig model. Food Funct 9:4153–4163CrossRefGoogle Scholar
  34. 34.
    Jones JD (1964) Lysine–arginine antagonism in the chick. J Nutr 84:313–321CrossRefGoogle Scholar
  35. 35.
    Austic RE, Scott RL (1975) Involvement of food intake in the lysine–arginine antagonism in chicks. J Nutr 105:1122–1131CrossRefGoogle Scholar
  36. 36.
    Mogensen CE, Solling (1977) Studies on renal tubular protein reabsorption: partial and near complete inhibition by certain amino acids. Scand J Clin Lab Invest 37:477–486CrossRefGoogle Scholar
  37. 37.
    Mogensen CE, Vittinghus E, Solling K (1979) Abnormal albumin excretion after two provocative renal tests in diabetes: physical exercise and lysine injection. Kidney Int 16:385–393CrossRefGoogle Scholar
  38. 38.
    Winterborn MH, Bradwell AR, Chesner IM, Jones GT (1987) The origin of proteinuria at high altitude. Postgrad Med J 63:179–181CrossRefGoogle Scholar
  39. 39.
    Racusen LC, Whelton A, Solez K (1985) Effects of lysine and other amino acids on kidney structure and function in the rat. Am J Pathol 120:436–442PubMedPubMedCentralGoogle Scholar
  40. 40.
    Wynn JL, Scumpia PO, Winfield RD, Delano MJ, Kelly-Scumpia K, Barker T, Ungaro R, Levy O, Moldawer LL (2008) Defective innate immunity predisposes murine neonates to poor sepsis outcome but is reversed by TLR agonists. Blood 112:1750–1758CrossRefGoogle Scholar
  41. 41.
    Deshmukh HS, Liu Y, Menkiti OR, Mei J, Dai N, O’Leary CE, Oliver PM, Kolls JK, Weiser JN, Worthen GS (2014) The microbiota regulates neutrophil homeostasis and host resistance to Escherichia coli K1 sepsis in neonatal mice. Nat Med 20:524–530CrossRefGoogle Scholar
  42. 42.
    Munder M (2009) Arginase: an emerging key player in the mammalian immune system. Br J Pharmacol 158:638–651CrossRefGoogle Scholar
  43. 43.
    Munder M, Schneider H, Luckner C, Giese T, Langhans CD, Fuentes JM, Kropf P, Mueller I, Kolb A, Modolell M, Ho AD (2006) Suppression of T-cell functions by human granulocyte arginase. Blood 108:1627–1634CrossRefGoogle Scholar
  44. 44.
    Azzara A, Carulli G, Sbrana S, Rizzuti-Gullaci A, Minnucci S, Natale M, Ambrogi F (1995) Effects of lysine-arginine association on immune functions in patients with recurrent infections. Drugs Exp Clin Res 21:71–78PubMedGoogle Scholar
  45. 45.
    Martins MJ, Negrao MR, Hipolito-Reis C, Azevedo I (2001) Arginine and a polyarginine peptide inhibit alkaline phosphatase activity: possible consequences for cellular transport systems. Clin Biochem 34:435–437CrossRefGoogle Scholar
  46. 46.
    Pekarthy JM, Short J, Lansing AI, Lieberman I (1972) Function and control of liver alkaline phosphatase. J Biol Chem 247:1767–1774PubMedGoogle Scholar
  47. 47.
    Fishman W, Sie HG (1970) L-homoarginine; an inhibitor of serum “bone and liver” alkaline phosphatase. Clin Chim Acta 29:339–341CrossRefGoogle Scholar
  48. 48.
    Ryan WL, Wells IC (1964) Homocitrulline and homoarginine synthesis from lysine. Science 144:1122–1127CrossRefGoogle Scholar
  49. 49.
    Ryan WL, Johnson RJ, Dimari S (1969) Homoarginine synthesis by rat kidney. Arch Biochem Biophys 131:521–526CrossRefGoogle Scholar
  50. 50.
    Torricelli P, Fini M, Giavaresi G, Giardino R, Gnudi S, Nicolini A, Carpi A (2002) L-Arginine and L-lysine stimulation on cultured human osteoblasts. Biomed Pharmacother 56:492–497CrossRefGoogle Scholar
  51. 51.
    Yu GC, Lee DB (1987) Clinical disorders of phosphorus metabolism. West J Med 147:569–576PubMedPubMedCentralGoogle Scholar
  52. 52.
    Liao SF, Wang T, Regmi N (2015) Lysine nutrition in swine and the related monogastric animals: muscle protein biosynthesis and beyond. Springerplus 4:147CrossRefGoogle Scholar
  53. 53.
    Chromiak JA, Antonio J (2002) Use of amino acids as growth hormone-releasing agents by athletes. Nutrition 18:657–661CrossRefGoogle Scholar
  54. 54.
    Isidori A, Lo MA, Cappa M (1981) A study of growth hormone release in man after oral administration of amino acids. Curr Med Res Opin 7:475–481CrossRefGoogle Scholar
  55. 55.
    Corpas E, Blackman MR, Roberson R, Scholfield D, Harman SM (1993) Oral arginine-lysine does not increase growth hormone or insulin-like growth factor-I in old men. J Gerontol 48:M128–M133CrossRefGoogle Scholar
  56. 56.
    Floyd JC Jr, Fajans SS, Conn JW, Knopf RF, Rull J (1966) Insulin secretion in response to protein ingestion. J Clin Investig 45:1479–1486CrossRefGoogle Scholar
  57. 57.
    Kalogeropoulou D, LaFave L, Schweim K, Gannon MC, Nuttall FQ (2009) Lysine ingestion markedly attenuates the glucose response to ingested glucose without a change in insulin response. Am J Clin Nutr 90:314–320CrossRefGoogle Scholar
  58. 58.
    Hutton JC, Sener A, Malaisse WJ (1980) Interaction of branched chain amino acids and keto acids upon pancreatic islet metabolism and insulin secretion. J Biol Chem 255:7340–7346PubMedGoogle Scholar
  59. 59.
    Amitani M, Asakawa A, Amitani H, Inui A (2013) The role of leptin in the control of insulin-glucose axis. Front Neurosci 7:51CrossRefGoogle Scholar
  60. 60.
    Kuhara T, Ikeda S, Ohneda A, Sasaki Y (1991) Effects of intravenous infusion of 17 amino acids on the secretion of GH, glucagon, and insulin in sheep. Am J Physiol 260:E21–E26PubMedGoogle Scholar
  61. 61.
    Rocha DM, Faloona GR, Unger RH (1972) Glucagon-stimulating activity of 20 amino acids in dogs. J Clin Investig 51:2346–2351CrossRefGoogle Scholar
  62. 62.
    Gannon MC, Nuttall FQ (2010) Amino acid ingestion and glucose metabolism–a review. IUBMB Life 62:660–668CrossRefGoogle Scholar
  63. 63.
    Lemieux I, Lamarche B, Couillard C, Pascot A, Cantin B, Bergeron J, Dagenais GR, Despres JP (2001) Total cholesterol/HDL cholesterol ratio vs LDL cholesterol/HDL cholesterol ratio as indices of ischemic heart disease risk in men: the Quebec Cardiovascular Study. Arch Intern Med 161:2685–2692CrossRefGoogle Scholar
  64. 64.
    Fonnebo V, Dahl LB, Moe PJ, Ingebretsen OC (1991) Does VLDL-LDL-cholesterol in cord serum predict future level of lipoproteins? Acta Paediatr Scand 80:780–785CrossRefGoogle Scholar
  65. 65.
    Schmeisser DD, Kummerow FA, Baker DH (1983) Effect of excess dietary lysine on plasma lipids of the chick. J Nutr 113:1777–1783CrossRefGoogle Scholar
  66. 66.
    Sanchez A, Rubano DA, Shavlik GW, Hubbard R, Horning MC (1988) Cholesterolemic effects of the lysine/arginine ratio in rabbits after initial early growth. Arch Latinoam Nutr 38:229–238PubMedGoogle Scholar
  67. 67.
    Lonovics J, Devitt P, Watson LC, Rayford PL, Thompson JC (1981) Pancreatic polypeptide. A review. Arch Surg 116:1256–1264CrossRefGoogle Scholar
  68. 68.
    Bard JA, Walker MW, Branchek TA, Weinshank RL (1995) Cloning and functional expression of a human Y4 subtype receptor for pancreatic polypeptide, neuropeptide Y, and peptide YY. J Biol Chem 270:26762–26765CrossRefGoogle Scholar
  69. 69.
    Batterham RL, Le Roux CW, Cohen MA, Park AJ, Ellis SM, Patterson M, Frost GS, Ghatei MA, Bloom SR (2003) Pancreatic polypeptide reduces appetite and food intake in humans. J Clin Endocrinol Metab 88:3989–3992CrossRefGoogle Scholar
  70. 70.
    Yin J, Li Y, Han H, Zheng J, Wang L, Ren W, Chen S, Wu F, Fang R, Huang X, Li C, Tan B, Xiong X, Zhang Y, Liu G, Yao J, Li T, Yin Y (2017) Effects of Lysine deficiency and Lys-Lys dipeptide on cellular apoptosis and amino acids metabolism. Mol Nutr Food Res 61:1600754CrossRefGoogle Scholar
  71. 71.
    Imamura W, Yoshimura R, Takai M, Yamamura J, Kanamoto R, Kato H (2013) Adverse effects of excessive leucine intake depend on dietary protein intake: a transcriptomic analysis to identify useful biomarkers. J Nutr Sci Vitaminol (Tokyo) 59:45–55CrossRefGoogle Scholar

Copyright information

© Her Majesty the Queen in Right of Canada, represented by the Minister of Health 2018

Authors and Affiliations

  1. 1.Nutrition Research Division, Bureau of Nutritional Sciences, Food Directorate, Health Products and Food BranchHealth CanadaOttawaCanada
  2. 2.Food and Nutrition Science Program, Department of ChemistryCarleton UniversityOttawaCanada

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