Advertisement

European Journal of Applied Physiology

, Volume 96, Issue 4, pp 363–369 | Cite as

Regular endurance training reduces the exercise induced HIF-1α and HIF-2α mRNA expression in human skeletal muscle in normoxic conditions

  • Carsten LundbyEmail author
  • Max Gassmann
  • Henriette Pilegaard
Original Article

Abstract

Regular exercise induces a variety of adaptive responses that enhance the oxidative and metabolic capacity of human skeletal muscle. Although the physiological adjustments of regular exercise have been known for decades, the underlying mechanisms are still unclear. The hypoxia inducible factors 1 and 2 (HIFs) are clearly related heterodimeric transcription factors that consist of an oxygen-depended α-subunit and a constitutive β-subunit. With hypoxic exposure, HIF-1α and HIF-2α protein are stabilized. Upon heterodimerization, HIFs induce the transcription of a variety of genes including erythropoietin (EPO), transferrin and its receptor, as well as vascular endothelial growth factor (VEGF) and its receptor. Considering that several of these genes are also induced with exercise, we tested the hypothesis that the mRNA level of HIF-1α and HIF-2α subunits increases with a single exercise bout, and that this response is blunted with training. We obtained muscle biopsies from a trained (5 days/week during 4 weeks) and untrained leg from the same human subject before, immediately after, and during the recovery from a 3 h two-legged knee extensor exercise bout, where the two legs exercised at the same absolute workload. In the untrained leg, the exercise bout induced an increase (P<0.05) in HIF-1α fold and HIF-2α fold mRNA at 6 h of recovery. In contrast, HIF-1α and HIF-2α mRNA levels were not altered at any time point in the trained leg. Obviously, HIF-1α and HIF-2α mRNA levels are transiently increased in untrained human skeletal muscle in response to an acute exercise bout, but this response is blunted after exercise training. We propose that HIFs expression is upregulated with exercise and that it may be an important transcription factor that regulates adaptive gene responses to exercise.

Keywords

Vascular Endothelial Growth Factor Human Skeletal Muscle Exercise Bout Vascular Endothelial Growth Factor Gene Prior Optimization 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Notes

Acknowledgments

Lara Ogunshola, Stephan Keller, and Bengt Saltin are thanked for their fruitful discussions throughout the study and while preparing the manuscript. Max Gassmann is founded by the Swiss National Science Foundation. Henriette Pilegaard and Carsten Lundby is founded by The Danish Medical Research Council and The Danish Natural Science Research Council.

References

  1. Ameln H, Gustafsson T, Sundberg CJ, Okamoto K, Jansson E, Poellinger L, Makino Y (2005) Physiological activation of hypoxia inducible factor-1 in human skeletal muscle. FASEB J 19:1009–1011PubMedGoogle Scholar
  2. Andersen P, Saltin B (1985) Maximal perfusion of skeletal muscle in man. J Physiol 366:233–249PubMedGoogle Scholar
  3. Bergstrøm J (1962) Muscle electrolytes in man. Scan J Clin Lab Invest 68:1–110Google Scholar
  4. Chomczynski P, Sacchi N (1987) Single-step method of RNA isolation by acid guanidinium thiocyanate–phenol–chloroform extraction. Anal Biochem 162:156–159PubMedCrossRefGoogle Scholar
  5. Connett RJ, Gayeski TE, Honig CR (1984) Lactate accumulation in fully aerobic, working, dog gracilis muscle. Am J Physiol 246:H120–H128PubMedGoogle Scholar
  6. Ehrnborg C, Lange KH, Dall R, Christiansen JS, Lundberg PA, Baxter RC, Boroujerdi MA, Bengtsson BA, Healey ML, Pentecost C, Longobardi S, Napoli R, Rosen T (2003) The growth hormone/insulin-like growth factor-I axis hormones and bone markers in elite athletes in response to a maximum exercise test. J Clin Endocrinol Metab 88:394–401PubMedCrossRefGoogle Scholar
  7. Feldser D, Agani F, Iyer NV, Pak B, Ferreira G, Semenza GL (1999) Reciprocal positive regulation of hypoxia-inducible factor 1alpha and insulin-like growth factor 2. Cancer Res 59:3915–3918PubMedGoogle Scholar
  8. Gladden L (1996) Lactate transport and exchange during exercise. In: Handbook of physiology. Exercise: regulation and integration of multiple systems. Am Physiol Soc, Bethesda, pp 614–649Google Scholar
  9. Gorlach A, Diebold I, Schini-Kerth VB, Berchner-Pfannschmidt U, Roth U, Brandes RP, Kietzmann T, Busse R (2001) Thrombin activates the hypoxia-inducible factor-1 signaling pathway in vascular smooth muscle cells: role of the p22(phox)-containing NADPH oxidase. Circ Res 89:47–54PubMedCrossRefGoogle Scholar
  10. Gustafsson T, Puntschart A, Kaijser L, Jansson E, Sundberg CJ (1999) Exercise-induced expression of angiogenesis-related transcription and growth factors in human skeletal muscle. Am J Physiol 276:H679–H685PubMedGoogle Scholar
  11. Hellwig-Burgel T, Rutkowski K, Metzen E, Fandrey J, Jelkmann W (1999) Interleukin-1beta and tumor necrosis factor-alpha stimulate DNA binding of hypoxia-inducible factor-1. Blood 94:1561–1567PubMedGoogle Scholar
  12. Honig CR, Gayeski TE, Clark A Jr, Clark PA (1991) Arteriovenous oxygen diffusion shunt is negligible in resting and working gracilis muscles. Am J Physiol 261:H2031–H2043PubMedGoogle Scholar
  13. Hopfl G, Ogunshola O, Gassmann M (2003) Hypoxia and high altitude. The molecular response. Adv Exp Med Biol 543:89–115PubMedGoogle Scholar
  14. Hopfl G, Ogunshola O, Gassmann M (2004) HIFs and tumors—causes and consequences. Am J Physiol Regul Integr Comp Physiol 286:R608–R623PubMedGoogle Scholar
  15. Jewell UR, Kvietikova I, Scheid A, Bauer C, Wenger RH, Gassmann M (2001) Induction of HIF-1alpha in response to hypoxia is instantaneous. FASEB J 15:1312–1314PubMedGoogle Scholar
  16. Joyner MJ, Tschakovsky ME (2003) Nitric oxide and physiologic vasodilation in human limbs: where do we go from here? Can J Appl Physiol 28:475–490PubMedGoogle Scholar
  17. Koval JA, DeFronzo RA, O’Doherty RM, Printz R, Ardehali H, Granner DK, Mandarino LJ (1998) Regulation of hexokinase II activity and expression in human muscle by moderate exercise. Am J Physiol 274:E304–E308PubMedGoogle Scholar
  18. Laughner E, Taghavi P, Chiles K, Mahon PC, Semenza GL (2001) HER2 (neu) signaling increases the rate of hypoxia-inducible factor 1alpha (HIF-1alpha) synthesis: novel mechanism for HIF-1-mediated vascular endothelial growth factor expression. Mol Cell Biol 21:3995–4004PubMedCrossRefGoogle Scholar
  19. Lundby C, Nordsborg N, Kusuhara K, Kristensen KM, Neufer PD, Pilegaard H (2005) Gene expression in human skeletal muscle: alternative normalization method and effect of repeated biopsies. Eur J Appl Physiol 1–10Google Scholar
  20. Mole PA, Chung Y, Tran TK, Sailasuta N, Hurd R, Jue T (1999) Myoglobin desaturation with exercise intensity in human gastrocnemius muscle. Am J Physiol 277:R173–R180PubMedGoogle Scholar
  21. Mourtzakis M, Gonzalez-Alonso J, Graham TE, Saltin B (2004) Hemodynamics and O2 uptake during maximal knee extensor exercise in untrained and trained human quadriceps muscle: effects of hyperoxia. J Appl Physiol 97:1796–1802PubMedCrossRefGoogle Scholar
  22. Ookawara T, Suzuk K, Haga S, Ha S, Chung KS, Toshinai K, Hamaoka T, Katsumura T, Takemasa T, Mizuno M, Hitomi Y, Kizaki T, Suzuki K, Ohno H (2002) Transcription regulation of gene expression in human skeletal muscle in response to endurance training. Res Commun Mol Pathol Pharmacol 111:41–54PubMedGoogle Scholar
  23. Ostrowski K, Rohde T, Asp S, Schjerling P, Pedersen BK (1999) Pro- and anti-inflammatory cytokine balance in strenuous exercise in humans. J Physiol 515(Pt 1):287–291PubMedCrossRefGoogle Scholar
  24. Pilegaard H, Ordway GA, Saltin B, Neufer PD (2000) Transcriptional regulation of gene expression in human skeletal muscle during recovery from exercise. Am J Physiol Endocrinol Metab 279:E806–E814PubMedGoogle Scholar
  25. Pilegaard H, Saltin B, Neufer PD (2003) Exercise induces transient transcriptional activation of the PGC-1alpha gene in human skeletal muscle. J Physiol 546:851–858PubMedCrossRefGoogle Scholar
  26. Rennie MJ, Park DM, Sulaiman WR (1976) Uptake and release of hormones and metabolites by tissues of exercising leg in man. Am J Physiol 231:967–973PubMedGoogle Scholar
  27. Richardson RS, Noyszewski EA, Kendrick KF, Leigh JS, Wagner PD (1995) Myoglobin O2 desaturation during exercise. Evidence of limited O2 transport. J Clin Invest 96:1916–1926PubMedCrossRefGoogle Scholar
  28. Richardson RS, Leigh JS, Wagner PD, Noyszewski EA (1999a) Cellular PO2 as a determinant of maximal mitochondrial O(2) consumption in trained human skeletal muscle. J Appl Physiol 87:325–331Google Scholar
  29. Richardson RS, Wagner H, Mudaliar SR, Henry R, Noyszewski EA, Wagner PD (1999b) Human VEGF gene expression in skeletal muscle: effect of acute normoxic and hypoxic exercise. Am J Physiol 277:H2247–H2252Google Scholar
  30. Richard DE, Berra E, Pouyssegur J (2000) Nonhypoxic pathway mediates the induction of hypoxia-inducible factor 1alpha in vascular smooth muscle cells. J Biol Chem 275:26765–26771PubMedCrossRefGoogle Scholar
  31. Richardson RS, Newcomer SC, Noyszewski EA (2001) Skeletal muscle intracellular PO(2) assessed by myoglobin desaturation: response to graded exercise. J Appl Physiol 91:2679–2685PubMedGoogle Scholar
  32. Saltin B, Gollnick PD (1983) Skeletal muscle adaptability: significance for metabolism and performance. In: Handbook of physiology. Exercise: regulation and integration of multiple systems, Am Physiol Soc, Bethesda, pp 555–631Google Scholar
  33. Stroka DM, Burkhardt T, Desbaillets I, Wenger RH, Neil DA, Bauer C, Gassmann M, Candinas D (2001) HIF-1 is expressed in normoxic tissue and displays an organ-specific regulation under systemic hypoxia. FASEB J 15:2445–2453PubMedGoogle Scholar
  34. Vogt M, Puntschart A, Geiser J, Zuleger C, Billeter R, Hoppeler H (2001) Molecular adaptations in human skeletal muscle to endurance training under simulated hypoxic conditions. J Appl Physiol 91:173–182PubMedGoogle Scholar
  35. Wiener CM, Booth G, Semenza GL (1996) In vivo expression of mRNAs encoding hypoxia-inducible factor 1. Biochem Biophys Res Commun 225:485–488PubMedCrossRefGoogle Scholar
  36. Wiesener MS, Jürgensen JS, Rosenberger C, Scholze CK, Horstrup JH, Warnecke C, Mandriota S, Bechmann I, Frei UA, Pugh CW, Ratcliffe PJ, Bachmann S, Maxwell PH, Eckardt KU (2003) Widespread hypoxia-inducible expression of HIF-2alpha in distinct cell populations of different organs. FASEB J 17:271–273PubMedGoogle Scholar
  37. Willimas RS, Neufer PD (1996) Regulation of gene expression in skeletal muscle by contractile activity. In: Rowell LSJ (ed) The handbook of physiology. Exercise: regulation, integration of multiple systems. Oxford University Press, New York, pp 1124–1150Google Scholar
  38. Zelzer E, Levy Y, Kahana C, Shilo BZ, Rubinstein M, Cohen B (1998) Insulin induces transcription of target genes through the hypoxia-inducible factor HIF-1alpha/ARNT. EMBO J 17:5085–5094PubMedCrossRefGoogle Scholar
  39. Zhong H, Chiles K, Feldser D, Laughner E, Hanrahan C, Georgescu MM, Simons JW, Semenza GL (2000) Modulation of hypoxia-inducible factor 1alpha expression by the epidermal growth factor/phosphatidylinositol 3-kinase/PTEN/AKT/FRAP pathway in human prostate cancer cells: implications for tumor angiogenesis and therapeutics. Cancer Res 60:1541–1545PubMedGoogle Scholar

Copyright information

© Springer-Verlag 2005

Authors and Affiliations

  • Carsten Lundby
    • 1
    • 2
    • 3
    Email author
  • Max Gassmann
    • 3
  • Henriette Pilegaard
    • 1
    • 4
  1. 1.Copenhagen Muscle Research CentreCopenhagenDenmark
  2. 2.RigshospitaletCopenhagenDenmark
  3. 3.Vetsuisse Faculty and Zürich Center for Integrative Human Physiology (ZIPH)University of ZürichZürichSwitzerland
  4. 4.Institute of Molecular Biology and Physiology, The August Krogh Institute University of CopenhagenCopenhagenDenmark

Personalised recommendations