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Analytical and Bioanalytical Chemistry

, Volume 396, Issue 3, pp 1167–1176 | Cite as

1H NMR-based metabolomics approach for exploring urinary metabolome modifications after acute and chronic physical exercise

  • C. Enea
  • F. Seguin
  • J. Petitpas-Mulliez
  • N. Boildieu
  • N. Boisseau
  • N. Delpech
  • V. Diaz
  • M. Eugène
  • B. DuguéEmail author
Original Paper

Abstract

Metabolomics is a comprehensive method for metabolite assessment that involves measuring the overall metabolic signature of biological samples. We used this approach to investigate biochemical changes due to acute and chronic physical exercise. Twenty-two women using identical oral contraceptives were segregated into an untrained (n = 10) or trained (n = 12) group depending on their physical training background. The subjects performed two exercises in a randomized order: a prolonged exercise test (75% of their \( \mathop V\limits^\cdot {{\text{O}}_{2\,\;\max }} \) until exhaustion) and a short-term, intensive exercise test (short-term, intensive exercise anaerobic test). Urine specimens were collected before and 30 min after each test. The samples were analyzed by 1H NMR spectroscopy, and multivariate statistical techniques were utilized to process the data. Distinguishing characteristics were observed only in the urine profiles of specimens collected before vs. 30 min after the short-term, intensive exercise test. The metabolites responsible for such changes were creatinine, lactate, pyruvate, alanine, β-hydroxybutyrate, acetate, and hypoxanthine. In both groups, the excretion of lactate, pyruvate, alanine, β-hydroxybutyrate, and hypoxanthine increased similarly after the completion of the short-term, intensive exercise test (p < 0.03). However, acetate excretion increased to a lesser extent in trained than in untrained subjects (p < 0.05). In conclusion, metabolomics is a promising tool in order to gain insight into physiological status and to clarify the changes induced by short-term, intense physical exercise.

Keywords

Biological variation Metabolomics Physical exercise Pre-analytical factor Urine analysis Young women 

Notes

Acknowledgment

This research was supported by the World Anti-Doping Agency.

References

  1. 1.
    Dugué B, Leppanen E, Grasbeck R (1996) Preanalytical factors and the measurement of cytokines in human subjects. Int J Clin Lab Res 26:99–105CrossRefGoogle Scholar
  2. 2.
    Enea C, Boisseau N, Diaz V, Dugué B (2008) Biological factors and the determination of androgens in female subjects. Steroids 73:1203–1216CrossRefGoogle Scholar
  3. 3.
    Nicholson JK, Lindon JC, Holmes E (1999) ‘Metabonomics’: understanding tne metabolic responses of living systems to pathophysiological stimuli via multivariate statistical analysis of biological NMR spectroscopic data. Xenobiotica 29:1181–1189CrossRefGoogle Scholar
  4. 4.
    Shockcor JP, Holmes E (2002) Metabonomic applications in toxicity screening and disease diagnosis. Current Topics in Medicinal Chemistry 2:35–51CrossRefGoogle Scholar
  5. 5.
    Vandewalle H, Pérès G, Heller J, Monod H (1985) All out anaerobic capacity tests on cycle ergometers, A comparative study on men and women. Eur J Appl Physiol Occup Physiol 54:222–229CrossRefGoogle Scholar
  6. 6.
    Wishart DS et al (2007) A knowledge for the human metabolome. Nucleic Acids Res 35:521–526 http://www.ncbi.nlm.nih.gov/pubmed/17202168 CrossRefGoogle Scholar
  7. 7.
    Lundberg P, Vogel T, Malusek A, Lundquist PO, Cohen L, Dahlqvist O (2005) The magnetic resonance metabolomics database. ESMRMB 2005, Basel, Switzerland. http://www.liu.se/hu/mdl/main/
  8. 8.
    Zuppi C, Messana I, Forni F, Rossi C, Pennacchietti L, Ferrari F, Giardina B (1997) 1H NMR spectra of normal urines: reference ranges of the major metabolites. Clin Chim Acta 265:85–97CrossRefGoogle Scholar
  9. 9.
    Dugué B, Leppänen E, Gräsbeck G (1998) Are the preanalytical factors underestimated in clinical studies? Clin Chem Lab Med 36:811CrossRefGoogle Scholar
  10. 10.
    Dugué B, Leppänen E, Gräsbeck G (1999) Preanalytical factors (biological variation) and the measurement of serum soluble intercellular adhesion molecule-1 in humans: influence of the time of day, food intake, and physical and psychological stress. Clin Chem 45:1543–1547Google Scholar
  11. 11.
    Smith J, Hill D (1991) Contribution of energy systems during a Wingate power test. Br J Sports Med 25:196–199CrossRefGoogle Scholar
  12. 12.
    Le Moyec L, Pruna A, Eugene M, Bedrossian J, Idatte JM, Huneau JF, Tome D (1993) Proton nuclear magnetic resonance spectroscopy of urine and plasma in renal transplantation follow-up. Nephron 65:433–439CrossRefGoogle Scholar
  13. 13.
    Granier P, Mercier B, Mercier J, Anselme F, Préfaut C (1995) Aerobic and anaerobic contribution to Wingate test performance in sprint and middle-distance runners. Eur J Appl Physiol Occup Physiol 70:58–65CrossRefGoogle Scholar
  14. 14.
    Poortmans J, Vancalck B (1978) Renal glomerular and tubular impairment during strenuous exercise in young women. Eur J Clin Invest 8:175–178CrossRefGoogle Scholar
  15. 15.
    Mayersohn M, Conrad K, Achari R (1983) The influence of a cooked meat meal on creatinine plasma concentration and creatinine clearance. Br J Clin Pharmacol 15:227–230Google Scholar
  16. 16.
    Lukaski H (1996) Estimation of muscle mass. Human Kinetics, Champaign, pp 109–128Google Scholar
  17. 17.
    Stathis C, Febbraio M, Carey M, Snow R (1994) Influence of sprint training on human skeletal muscle purine nucleotide metabolism. J Appl Physiol 76:1802–1809Google Scholar
  18. 18.
    Tullson P, Bangsbo J, Hellsten Y, Richter E (1995) IMP metabolism in human skeletal muscle after exhaustive exercise. J Appl Physiol 78:146–152Google Scholar
  19. 19.
    Zhao S, Snow R, Stathis C, Febbraio M, Carey M (2000) Muscle adenine nucleotide metabolism during and in recovery from maximal exercise in humans. J Appl Physiol 88:1513–1519Google Scholar
  20. 20.
    Ihara H, Shino Y, Morita Y, Kawaguchi E, Hashizume N, Yoshida M (2001) Is skeletal muscle damaged by the oxidative stress following anaerobic exercise? J Clin Lab Anal 15:239–243CrossRefGoogle Scholar
  21. 21.
    Feet B, Yu X, Rootwelt T, Oyasaeter S, Saugstad O (1997) Effects of hypoxemia and reoxygenation with 21% or 100% oxygen in newborn piglets: extracellular hypoxanthine in cerebral cortex and femoral muscle. Crit Care Med 25:1384–1391CrossRefGoogle Scholar
  22. 22.
    Hellsten-Westing Y, Norman B, Sjödin B (1993) The effect of high-intensity training on purine metabolism in man. Acta Physiol Scand 149:405–412CrossRefGoogle Scholar
  23. 23.
    Stathis C, Carey M, Hayes A, Garnham A, Snow R (2006) Sprint training reduces urinary purine loss following intense exercise in humans. Appl Physiol Nutr Metab 31:702–708CrossRefGoogle Scholar
  24. 24.
    Thomas C, Perrey S, Lambert K, Hugon G, Mornet D, Mercier J (2005) Monocarboxylate transporters, blood lactate removal after supramaximal exercise, and fatigue indexes in humans. J Appl Physiol 98:804–809CrossRefGoogle Scholar
  25. 25.
    Thomas C, Sirvent P, Perrey S, Raynaud E, Mercier J (2004) Relationships between maximal muscle oxidative capacity and blood lactate removal after supramaximal exercise and fatigue indexes in humans. J Appl Physiol 97:2132–2138CrossRefGoogle Scholar
  26. 26.
    Freikman I, Amer J, Cohen J, Ringel I, Fibach E (2008) Oxidative stress causes membrane phospholipid rearrangement and shedding from RBC membranes—an NMR study. Biochim Biophys Acta 1778:2388–2394CrossRefGoogle Scholar
  27. 27.
    Bassenge E, Sommer O, Schwemmer M, Bünger R (2000) Antioxidant pyruvate inhibits cardiac formation of reactive oxygen species through changes in redox state. Am J Physiol Heart Circ Physiol 279:2431–2438Google Scholar
  28. 28.
    Hauet T, Eugene M (2008) A new approach in organ preservation: potential role of new polymers. Kidney Int 74:998–1003CrossRefGoogle Scholar
  29. 29.
    Groussard C, Rannou-Bekono F, Machefer G, Chevanne M, Vincent S, Sergent O (2003) Changes in blood lipid peroxidation markers and antioxidants after a single sprint anaerobic exercise. Eur J Appl Physiol 89:14–20CrossRefGoogle Scholar
  30. 30.
    Bloomer R, Fisher-Wellman K (2008) Blood oxidative stress biomarkers: influence of sex, exercise training status, and dietary intake. Gend Med 5:218–228CrossRefGoogle Scholar
  31. 31.
    Finaud J, Lac G, Filaire E (2006) Oxidative stress: relationship with exercise and training. Sports Med 36:327–358CrossRefGoogle Scholar
  32. 32.
    Dohm G, Tapscott E, Kasperek G (1987) Protein degradation during endurance exercise and recovery. Med Sci Sports Exerc 19:166–171Google Scholar
  33. 33.
    Williams B, Chinkes D, Wolfe R (1998) Alanine and glutamine kinetics at rest and during exercise in humans. Med Sci Sports Exerc 30:1053–1058CrossRefGoogle Scholar
  34. 34.
    Felig P, Wahren J (1971) Amino acid metabolism in exercising man. J Clin Invest 50:2703–2714CrossRefGoogle Scholar
  35. 35.
    Yan B, Wang G, Lu H, Huang X, Liu Y, Zha W, Hao H, Zhang Y, Liu L, Gu S, Huang Q, Zheng Y, Sun J (2009) Metabolomic investigation into variation of endogenous metabolites in professional athletes subject to strength–endurance training. J Appl Physiol 106:531–538CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  • C. Enea
    • 1
  • F. Seguin
    • 2
  • J. Petitpas-Mulliez
    • 1
    • 3
  • N. Boildieu
    • 2
  • N. Boisseau
    • 1
  • N. Delpech
    • 1
  • V. Diaz
    • 1
    • 3
  • M. Eugène
    • 2
    • 3
  • B. Dugué
    • 1
    Email author
  1. 1.Laboratoire des Adaptations Physiologiques aux Activités Physiques (EA 3813), Faculté des Sciences du SportUniversité de PoitiersPoitiersFrance
  2. 2.INSERM, U927, Faculté de Médecine et de PharmacieUniversité de PoitiersPoitiersFrance
  3. 3.Service de PhysiologieCHU PoitiersPoitiersFrance

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