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Resistance exercise increases intramuscular NF-κb signaling in untrained males

European Journal of Applied Physiology volume 116, pages 2103–2111 (2016)Cite this article

Abstract

Purpose

The NF-κB signaling pathway regulates multiple cellular processes following exercise stress. This study aims to examine the effects of an acute lower-body resistance exercise protocol and subsequent recovery on intramuscular NF-κB signaling.

Methods

Twenty-eight untrained males were assigned to either a control (CON; n = 11) or exercise group (EX; n = 17) and completed a lower-body resistance exercise protocol consisting of the back squat, leg press, and leg extension exercises. Skeletal muscle microbiopsies were obtained from the vastus lateralis pre-exercise (PRE), 1-hour (1H), 5-hours (5H), and 48-hours (48H) post-resistance exercise. Multiplex signaling assay kits (EMD Millipore, Billerica, MA, USA) were used to quantify the total protein (TNFR1, c-Myc) or phosphorylation status of proteins belonging to the NF-κB signaling pathway (IKKa/b, IkBα, NF-κB) using multiplex protein assay. Repeated measures ANOVA analysis was used to determine the effects of the exercise bout on intramuscular signaling at each time point. Additionally, change scores were analyzed by magnitude based inferences to determine a mechanistic interpretation.

Results

Repeated measures ANOVA indicated a trend for a two-way interaction between the EX and CON Group (p = 0.064) for c-Myc post resistance exercise. Magnitude based inference analysis suggest a “Very Likely” increase in total c-Myc from PRE-5H and a “Likely” increase in IkBα phosphorylation from PRE-5H post-resistance exercise.

Conclusion

Results indicated that c-Myc transcription factor is elevated following acute intense resistance exercise in untrained males. Future studies should examine the role that post-resistance exercise NF-κβ signaling plays in c-Myc induction, ribosome biogenesis and skeletal muscle regeneration.

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Abbreviations

ANOVA:

Analysis of variance

CK:

Creatine kinase

ELISA:

Enzyme-linked immunosorbent assay

ERK1/2:

Extracellular-signal-regulated kinase 1 and 2

LDH:

Lactate dehydrogenase

mRNA:

Messenger ribonucleic acid

MAPK:

Mitogen activated protein kinase

mTOR:

Mammalian target of rapamycin

NF-κB:

Nuclear factor kappa beta

ROS:

Reactive oxygen species

TNF-α:

Tumor necrosis factor alpha

References

  • Arabi A, Wu S, Ridderstråle K et al (2005) c-Myc associates with ribosomal DNA and activate RNA polymerase I transcription. Nat Cell Biol 7:303–310

    Article  CAS  PubMed  Google Scholar 

  • Bakkar N, Guttridge DC (2010) NF-kappaB signaling: a tale of two pathways in skeletal myogenesis. Physiol Rev 90:495–511. doi:10.1152/physrev.00040.2009

    Article  CAS  PubMed  Google Scholar 

  • Batterham AM, Hopkins WG (2006) Making meaningful inferences about magnitudes. Int J Sports Physiol Perform 1(1):50–57

    Article  PubMed  Google Scholar 

  • Beck TW (2013) The importance of a priori sample size estimation in strength and conditioning research. J Strength Cond Res 27(8):2323–2337

    Article  PubMed  Google Scholar 

  • Chaillou T, Kirby TJ, McCarthy JJ (2014) Ribosome biogenesis: emerging evidence for a central role in the regulation of skeletal muscle mass. J Cell Physiol 229:1584–1594

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Coffey VG, Hawley JA (2007) The molecular bases of training adaptation. Sports Med 37(9):737–763

    Article  PubMed  Google Scholar 

  • Coffey VG, Zhong Z, Shield A et al (2006) Early signaling responses to divergent exercise stimuli in skeletal muscle from well-trained humans. FASEB J 20(1):190–192

    CAS  PubMed  Google Scholar 

  • Constantin-Teodosiu D, Casey A, Short A, Hultman E, Greenhaff P (1996) The effect of repeated muscle biopsy sampling on ATP and glycogen resynthesis following exercise in man. Eur J Appl Physiol Occup Physiol 73:186–190

    Article  CAS  PubMed  Google Scholar 

  • Dahlman JM, Bakkar N, He W, Guttridge DC (2010) NF-kappaB functions in stromal fibroblasts to regulate early postnatal muscle development. J Biol Chem 285:5479–5487. doi:10.1074/jbc.M109.075606

    Article  CAS  PubMed  Google Scholar 

  • Dang CV (1999) c-Myc target genes involved in cell growth, apoptosis, and metabolism. Mol Cell Biol 19:1–11

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Durham WJ, Li YP, Gerken E et al (2004) Fatiguing exercise reduces DNA binding activity of NF-kappaB in skeletal muscle nuclei. J Appl Physiol (1985) 97:1740–1745. doi:10.1152/japplphysiol.00088.2004

    Article  CAS  Google Scholar 

  • Gomez-Roman N, Grandori C, Eisenman RN, White RJ (2003) Direct activation of RNA polymerase III transcription by c-Myc. Nature 421:290–294

    Article  CAS  PubMed  Google Scholar 

  • Gonzalez AM, Hoffman JR, Townsend JR et al (2015) Association between myosin heavy chain protein isoforms and intramuscular anabolic signaling following resistance exercise in trained men. Physiol Rep 3(1):e12268

    Article  PubMed  PubMed Central  Google Scholar 

  • Grandori C, Gomez-Roman N, Felton-Edkins ZA et al (2005) c-Myc binds to human ribosomal DNA and stimulates transcription of rRNA genes by RNA polymerase I. Nat Cell Biol 7:311–318

    Article  CAS  PubMed  Google Scholar 

  • Green S, Salkind N, Akey T (2000) Methods for controlling type I error across multiple hypothesis tests. Using SPSS Windows Anal Underst Data 2:395–396

    Google Scholar 

  • Hacker H, Karin M (2006) Regulation and function of IKK and IKK-related kinases. Sci Stke 357:12–14

    Google Scholar 

  • Hayden MS, Ghosh S (2004) Signaling to NF-kappaB. Genes Dev 18:2195–2224. doi:10.1101/gad.1228704

    Article  CAS  PubMed  Google Scholar 

  • Ho RC, Hirshman MF, Li Y et al (2005) Regulation of IkappaB kinase and NF-kappaB in contracting adult rat skeletal muscle. Am J Physiol Cell Physiol 289:C794–C801. doi:10.1152/ajpcell.00632.2004

  • Hoffman J (2006) Norms for fitness, performance, and health. Human Kinetics, Champaign

    Google Scholar 

  • Hollander J, Fiebig R, Gore M, Ookawara T, Ohno H, Ji L (2001) Superoxide dismutase gene expression is activated by a single bout of exercise in rat skeletal muscle. Pfluegers Arch 442:426–434

    Article  CAS  Google Scholar 

  • Hopkins WG (2002) Probabilities of clinical or practical significance. Sportscience 6:201

    Google Scholar 

  • Hopkins W (2007) A spreadsheet for deriving a confidence interval, mechanistic inference and clinical inference from ap value. Sportscience 11:16–20

    Google Scholar 

  • Hughes K, Antonsson Å, Grundstrøm T (1998) Calmodulin dependence of NFκB activation. FEBS Lett 441:132–136

    Article  CAS  PubMed  Google Scholar 

  • Hughes K, Edin S, Antonsson A, Grundstrom T (2001) Calmodulin-dependent kinase II mediates T cell receptor/CD3- and phorbol ester-induced activation of IkappaB kinase. J Biol Chem 276:36008–36013. doi:10.1074/jbc.M106125200

    Article  CAS  PubMed  Google Scholar 

  • Hunter RB, Stevenson E, Koncarevic A, Mitchell-Felton H, Essig DA, Kandarian SC (2002) Activation of an alternative NF-kappaB pathway in skeletal muscle during disuse atrophy. FASEB J 16:529–538

    Article  CAS  PubMed  Google Scholar 

  • Hyldahl RD, Xin L, Hubal MJ, Moeckel-Cole S, Chipkin S, Clarkson PM (2011) Activation of nuclear factor-kappaB following muscle eccentric contractions in humans is localized primarily to skeletal muscle-residing pericytes. FASEB J 25:2956–2966. doi:10.1096/fj.10-177105

    Article  CAS  PubMed  Google Scholar 

  • Ji LL, Gomez-Cabrera MC, Steinhafel N, Vina J (2004) Acute exercise activates nuclear factor (NF)-kappaB signaling pathway in rat skeletal muscle. FASEB J 18:1499–1506. doi:10.1096/fj.04-1846com

  • Kiuchi N, Nakajima K, Ichiba M et al (1999) STAT3 is required for the gp130-mediated full activation of the c-myc gene. J Exp Med 189:63–73

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Kramer HF, Goodyear LJ (2007) Exercise, MAPK, and NF-kappaB signaling in skeletal muscle. J Appl Physiol (1985) 103:388–395. doi:10.1152/japplphysiol.00085.2007

  • Lee J, Burckart GJ (1998) Nuclear factor kappa B: important transcription factor and therapeutic target. J Clin Pharmacol 38:981–993

    Article  CAS  PubMed  Google Scholar 

  • Li H, Lin X (2008) Positive and negative signaling components involved in TNFα-induced NF-κB activation. Cytokine 41:1–8

    Article  PubMed  Google Scholar 

  • Li YP, Reid MB (2000) NF-kappaB mediates the protein loss induced by TNF-alpha in differentiated skeletal muscle myotubes. Am J Physiol Regul Integr Comp Physiol 279:R1165–R1170

    CAS  PubMed  Google Scholar 

  • Mahoney DJ, Safdar A, Parise G et al (2008) Gene expression profiling in human skeletal muscle during recovery from eccentric exercise. Am J Physiol Regul Integr Comp Physiol 294:R1901–R1910. doi:10.1152/ajpregu.00847.2007

    Article  CAS  PubMed  Google Scholar 

  • Malm C, Nyberg P, Engström M et al (2000) Immunological changes in human skeletal muscle and blood after eccentric exercise and multiple biopsies. J Physiol (Lond) 529:243–262

    Article  CAS  Google Scholar 

  • Malm C, Sjödin B, Sjöberg B et al (2004) Leukocytes, cytokines, growth factors and hormones in human skeletal muscle and blood after uphill or downhill running. J Physiol (Lond) 556:983–1000

    Article  CAS  Google Scholar 

  • Masuda M, Suzui M, Yasumatu R et al (2002) Constitutive activation of signal transducers and activators of transcription 3 correlates with cyclin D1 overexpression and may provide a novel prognostic marker in head and neck squamous cell carcinoma. Cancer Res 62:3351–3355

    CAS  PubMed  Google Scholar 

  • McKay BR, De Lisio M, Johnston AP et al (2009) Association of interleukin-6 signalling with the muscle stem cell response following muscle-lengthening contractions in humans. PLoS One 4:e6027

    Article  PubMed  PubMed Central  Google Scholar 

  • Mobley CB, Fox CD, Thompson RM et al (2016) Comparative effects of whey protein versus l-leucine on skeletal muscle protein synthesis and markers of ribosome biogenesis following resistance exercise. Amino Acids 48:733–750. doi:10.1007/s00726-015-2121-z

  • Møller AB, Vendelbo MH, Rahbek SK et al (2013) Resistance exercise, but not endurance exercise, induces IKKβ phosphorylation in human skeletal muscle of training-accustomed individuals. Pflügers Arch Eur J Physiol 465:1785–1795

    Article  Google Scholar 

  • Mourkioti F, Rosenthal N (2008) NF-κB signaling in skeletal muscle: prospects for intervention in muscle diseases. J Mol Med 86:747–759

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Nader GA, von Walden F, Liu C et al (2014) Resistance exercise training modulates acute gene expression during human skeletal muscle hypertrophy. J Appl Physiol (1985) 116:693–702. doi:10.1152/japplphysiol.01366.2013

    Article  CAS  Google Scholar 

  • Peake JM, Suzuki K, Wilson G et al (2005) Exercise-induced muscle damage, plasma cytokines, and markers of neutrophil activation. Med Sci Sports Exerc 37:737–745

    Article  CAS  PubMed  Google Scholar 

  • Pedersen BK, Toft AD (2000) Effects of exercise on lymphocytes and cytokines. Br J Sports Med 34:246–251

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Powers SK, Jackson MJ (2008) Exercise-induced oxidative stress: cellular mechanisms and impact on muscle force production. Physiol Rev 88:1243–1276. doi:10.1152/physrev.00031.2007

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Reid MB, Durham WJ (2002) Generation of reactive oxygen and nitrogen species in contracting skeletal muscle. Ann N Y Acad Sci 959:108–116

    Article  CAS  PubMed  Google Scholar 

  • Schmidt EV (1999) The role of c-myc in cellular growth control. Oncogene 18:2988–2896. doi:10.1038/sj.onc.1202751

  • Schmitz ML, Baeuerle PA (1991) The p65 subunit is responsible for the strong transcription activating potential of NF-kappa B. EMBO J 10:3805–3817

    CAS  PubMed  PubMed Central  Google Scholar 

  • Toft AD, Jensen LB, Bruunsgaard H et al (2002) Cytokine response to eccentric exercise in young and elderly humans. Am J Physiol Cell Physiol 283:C289–C295

    Article  CAS  PubMed  Google Scholar 

  • Townsend JR, Hoffman JR, Fragala MS et al (2016) A microbiopsy method for immunohistological and morphological analysis: a pilot study. Med Sci Sports Exerc 48:331–335. doi:10.1249/MSS.0000000000000772

    Article  CAS  PubMed  Google Scholar 

  • Van Riggelen J, Yetil A, Felsher DW (2010) MYC as a regulator of ribosome biogenesis and protein synthesis. Nat Rev Cancer 10:301–309

    Article  PubMed  Google Scholar 

  • Vella L, Caldow MK, Larsen AE et al (2012) Resistance exercise increases NF-kappaB activity in human skeletal muscle. Am J Physiol Regul Integr Comp Physiol 302:R667–R673. doi:10.1152/ajpregu.00336.2011

    Article  CAS  PubMed  Google Scholar 

  • Vissing K, McGee S, Farup J, Kjølhede T, Vendelbo M, Jessen N (2013) Differentiated mTOR but not AMPK signaling after strength vs endurance exercise in training-accustomed individuals. Scand J Med Sci Sports 23:355–366

    Article  CAS  PubMed  Google Scholar 

  • Wei Q, Paterson BM (2001) Regulation of MyoD function in the dividing myoblast. FEBS Lett 490:171–178

    Article  CAS  PubMed  Google Scholar 

  • Wellen KE, Hotamisligil GS (2005) Inflammation, stress, and diabetes. J Clin Invest 115:1111–1119. doi:10.1172/JCI25102

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • West DW, Baehr LM, Marcotte GR et al (2016) Acute resistance exercise activates rapamycin-sensitive and-insensitive mechanisms that control translational activity and capacity in skeletal muscle. J Physiol (Lond) 594:453–468

    Article  CAS  Google Scholar 

  • Whitelaw PF, Hesketh JE (1992) Expression of c-myc and c-fos in rat skeletal muscle. Evidence for increased levels of c-myc mRNA during hypertrophy. Biochem J 281(Pt 1):143–147

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Wilson A, Murphy MJ, Oskarsson T et al (2004) c-Myc controls the balance between hematopoietic stem cell self-renewal and differentiation. Genes Dev 18:2747–2763

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Xin L, Hyldahl RD, Chipkin SR, Clarkson PM (2014) A contralateral repeated bout effect attenuates induction of NF-kappaB DNA binding following eccentric exercise. J Appl Physiol (1985) 116:1473–1480. doi:10.1152/japplphysiol.00133.2013

    Article  CAS  Google Scholar 

  • Yamaoka S, Courtois G, Bessia C et al (1998) Complementation cloning of NEMO, a component of the IκB kinase complex essential for NF-κB activation. Cell 93:1231–1240

    Article  CAS  PubMed  Google Scholar 

  • Yaron A, Hatzubai A, Davis M et al (1998) Identification of the receptor component of the IκBα–ubiquitin ligase. Nature 396:590–594

    Article  CAS  PubMed  Google Scholar 

  • Yu BP (1994) Cellular defenses against damage from reactive oxygen species. Physiol Rev 74:139–162

    CAS  PubMed  Google Scholar 

  • Zaldivar F, Wang-Rodriguez J, Nemet D et al (2006) Constitutive pro-and anti-inflammatory cytokine and growth factor response to exercise in leukocytes. J Appl Physiol 100:1124–1133

    Article  CAS  PubMed  Google Scholar 

  • Zandi E, Rothwarf DM, Delhase M, Hayakawa M, Karin M (1997) The IκB kinase complex (IKK) contains two kinase subunits, IKKα and IKKβ, necessary for IκB phosphorylation and NF-κB activation. Cell 91:243–252

    Article  CAS  PubMed  Google Scholar 

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Authors and Affiliations

  1. Exercise and Nutrition Science Graduate Program, Lipscomb University, Nashville, TN, 37215, USA

    Jeremy R. Townsend

  2. Institute of Exercise Physiology and Wellness, University of Central Florida, Orlando, FL, 32816, USA

    Jeffrey R. Stout, Adam R. Jajtner, David D. Church, Kyle S. Beyer, Leonardo P. Oliveira, Michael B. La Monica, Joshua J. Riffe, Tyler W. D. Muddle, Kayla M. Baker, David H. Fukuda & Jay R. Hoffman

  3. Molecular and Applied Sciences Laboratory, School of Kinesiology, Auburn University, Auburn, AL, 36849, USA

    Michael D. Roberts

Authors
  1. Jeremy R. Townsend
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  2. Jeffrey R. Stout
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Correspondence to Jeffrey R. Stout.

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This study was not supported by extramural funding. We certify that this work has not previously been published/presented in any form.

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Communicated by William J. Kraemer.

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Townsend, J.R., Stout, J.R., Jajtner, A.R. et al. Resistance exercise increases intramuscular NF-κb signaling in untrained males. Eur J Appl Physiol 116, 2103–2111 (2016). https://doi.org/10.1007/s00421-016-3463-2

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  • DOI: https://doi.org/10.1007/s00421-016-3463-2

Keywords

  • Ribosome biogenesis
  • Immune function
  • Recovery
  • Muscle damage