IL-6 Induced STAT3 Signalling Is Associated with the Proliferation of Human Muscle Satellite Cells Following Acute Muscle Damage

Background Although the satellite cell (SC) is a key regulator of muscle growth during development and muscle adaptation following exercise, the regulation of human muscle SC function remains largely unexplored. STAT3 signalling mediated via interleukin-6 (IL-6) has recently come to the forefront as a potential regulator of SC proliferation. The early response of the SC population in human muscle to muscle-lengthening contractions (MLC) as mediated by STAT3 has not been studied. Methodology/Principal Findings Twelve male subjects (21±2 y; 83±12 kg) performed 300 maximal MLC of the quadriceps femoris at 180°•s−1 over a 55° range of motion with muscle samples (vastus lateralis) and blood samples (antecubital vein) taken prior to exercise (PRE), 1 hour (T1), 3 hours (T3) and 24 hours (T24) post-exercise. Cytoplasmic and nuclear fractions of muscle biopsies were purified and analyzed for total and phosphorylated STAT3 (p-STAT3) by western blot. p-STAT3 was detected in cytoplasmic fractions across the time course peaking at T24 (p<0.01 vs. PRE). Nuclear total and p-STAT3 were not detected at appreciable levels. However, immunohistochemical analysis revealed a progressive increase in the proportion of SCs expressing p-STAT3 with ∼60% of all SCs positive for p-STAT3 at T24 (p<0.001 vs. PRE). Additionally, cMyc, a STAT3 downstream gene, was significantly up-regulated in SCs at T24 versus PRE (p<0.05). Whole muscle mRNA analysis revealed induction of the STAT3 target genes IL-6, SOCS3, cMyc (peaking at T3, p<0.05), IL-6Rα and GP130 (peaking at T24, p<0.05). In addition, Myf5 mRNA was up-regulated at T24 (p<0.05) with no appreciable change in MRF4 mRNA. Conclusions/Significant Findings We demonstrate that IL-6 induction of STAT3 signaling occurred exclusively in the nuclei of SCs in response to MLC. An increase in the number of cMyc+ SCs indicated that human SCs were induced to proliferate under the control of STAT3 signaling.

[1]  Volker Seifert,et al.  Inhibition of the JAK-2/STAT3 signaling pathway impedes the migratory and invasive potential of human glioblastoma cells , 2011, Journal of Neuro-Oncology.

[2]  D. Frank,et al.  Microtubule-Targeted Chemotherapeutic Agents Inhibit Signal Transducer and Activator of Transcription 3 (STAT3) Signaling , 2010, Molecular Pharmacology.

[3]  M. Tarnopolsky,et al.  Satellite cell number and cell cycle kinetics in response to acute myotrauma in humans: immunohistochemistry versus flow cytometry , 2010, The Journal of physiology.

[4]  J. Ponce-González,et al.  Muscle hypertrophy and increased expression of leptin receptors in the musculus triceps brachii of the dominant arm in professional tennis players , 2010, European Journal of Applied Physiology.

[5]  D. Schaffer,et al.  Relative roles of TGF-β1 and Wnt in the systemic regulation and aging of satellite cell responses , 2009, Aging cell.

[6]  Per Aagaard,et al.  Molecular aging and rejuvenation of human muscle stem cells , 2009, EMBO molecular medicine.

[7]  N. Reich.  STAT3 Revs Up the Powerhouse , 2009, Science Signaling.

[8]  M. Tarnopolsky,et al.  Association of Interleukin-6 Signalling with the Muscle Stem Cell Response Following Muscle-Lengthening Contractions in Humans , 2009, PloS one.

[9]  M. Tarnopolsky,et al.  Co‐expression of IGF‐1 family members with myogenic regulatory factors following acute damaging muscle‐lengthening contractions in humans , 2008, The Journal of physiology.

[10]  M. Febbraio,et al.  Muscle as an endocrine organ: focus on muscle-derived interleukin-6. , 2008, Physiological reviews.

[11]  M. Tarnopolsky,et al.  Hepatocyte growth factor (HGF) and the satellite cell response following muscle lengthening contractions in humans , 2008, Muscle & nerve.

[12]  J. Petrella,et al.  Potent myofiber hypertrophy during resistance training in humans is associated with satellite cell-mediated myonuclear addition: a cluster analysis. , 2008, Journal of applied physiology.

[13]  Thomas D. Schmittgen,et al.  Analyzing real-time PCR data by the comparative CT method , 2008, Nature Protocols.

[14]  D. Frank,et al.  Microtubule-targeted chemotherapeutic agents inhibit STAT3 signaling in breast cancer , 2008 .

[15]  H. Langberg,et al.  Myofibre damage in human skeletal muscle: effects of electrical stimulation versus voluntary contraction , 2007, The Journal of physiology.

[16]  G. Bassez,et al.  Muscle satellite cells and endothelial cells: close neighbors and privileged partners. , 2007, Molecular biology of the cell.

[17]  D. Cameron-Smith,et al.  STAT3 signaling is activated in human skeletal muscle following acute resistance exercise. , 2007, Journal of applied physiology.

[18]  David J. Kosek,et al.  Efficacy of myonuclear addition may explain differential myofiber growth among resistance-trained young and older men and women. , 2006, American journal of physiology. Endocrinology and metabolism.

[19]  J. Bromberg,et al.  Touched and Moved by STAT3 , 2006, Science's STKE.

[20]  M. Kjaer,et al.  Enhanced satellite cell proliferation with resistance training in elderly men and women , 2006, Scandinavian journal of medicine & science in sports.

[21]  R. Wiswell,et al.  Satellite cell numbers in young and older men 24 hours after eccentric exercise , 2006, Muscle & nerve.

[22]  F. Walsh,et al.  Myostatin: a modulator of skeletal-muscle stem cells. , 2005, Biochemical Society transactions.

[23]  A. Steensberg,et al.  Interleukin‐6 receptor expression in contracting human skeletal muscle: regulating role of IL‐6 , 2005, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[24]  M. Tarnopolsky,et al.  Pathology of skeletal muscle in mitochondrial disorders. , 2004, Mitochondrion.

[25]  L. Andersen,et al.  The effects of heavy resistance training and detraining on satellite cells in human skeletal muscles , 2004, The Journal of physiology.

[26]  Alla V. Ivanova,et al.  STRA13 interacts with STAT3 and modulates transcription of STAT3-dependent targets. , 2004, Journal of molecular biology.

[27]  H. Langberg,et al.  Changes in satellite cells in human skeletal muscle after a single bout of high intensity exercise , 2004, The Journal of physiology.

[28]  M. Febbraio,et al.  Skeletal myocytes are a source of interleukin‐6 mRNA expression and protein release during contraction: evidence of fiber type specificity , 2004, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[29]  F. Booth,et al.  Insulin-like growth factor 1 and muscle growth: implication for satellite cell proliferation , 2004, The Proceedings of the Nutrition Society.

[30]  D. Harrison,et al.  The JAK/STAT signaling pathway , 2004, Journal of Cell Science.

[31]  A. Steensberg,et al.  Interleukin-6 production by contracting human skeletal muscle: autocrine regulation by IL-6. , 2003, Biochemical and biophysical research communications.

[32]  M. Rudnicki,et al.  Looking back to the embryo: defining transcriptional networks in adult myogenesis , 2003, Nature Reviews Genetics.

[33]  C. Denis,et al.  Effects of endurance training on satellite cell frequency in skeletal muscle of old men , 2003, Muscle & nerve.

[34]  E. Senba,et al.  In Vivo Activation of STAT3 Signaling in Satellite Cells and Myofibers in Regenerating Rat Skeletal Muscles , 2002, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[35]  V. Edgerton,et al.  New Fiber Formation in the Interstitial Spaces of Rat Skeletal Muscle During Postnatal Growth , 2002, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[36]  F. Booth,et al.  Multiple signaling pathways mediate LIF-induced skeletal muscle satellite cell proliferation. , 2002, American journal of physiology. Cell physiology.

[37]  M. Masuda,et al.  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. , 2002, Cancer research.

[38]  M. Tarnopolsky,et al.  Contraction-induced muscle damage is unaffected by vitamin E supplementation. , 2002, Medicine and science in sports and exercise.

[39]  D. Levy,et al.  What does Stat3 do? , 2002, The Journal of clinical investigation.

[40]  F. López‐Soriano,et al.  Tumor necrosis factor-α exerts interleukin-6-dependent and -independent effects on cultured skeletal muscle cells , 2002 .

[41]  J. Fridén,et al.  Eccentric exercise-induced injuries to contractile and cytoskeletal muscle fibre components. , 2001, Acta physiologica Scandinavica.

[42]  B. Saltin,et al.  Production of interleukin‐6 in contracting human skeletal muscles can account for the exercise‐induced increase in plasma interleukin‐6 , 2000, The Journal of physiology.

[43]  K. Yarasheski,et al.  Myofibrillar disruption following acute concentric and eccentric resistance exercise in strength-trained men. , 2000, Canadian journal of physiology and pharmacology.

[44]  T. Naka,et al.  Negative regulation of cytokine signaling: STAT-induced STAT inhibitor. , 1999, Trends in biochemical sciences.

[45]  T. Hirano,et al.  STAT3 Is Required for the gp130-mediated Full Activation of the c-myc Gene , 1999, The Journal of experimental medicine.

[46]  Chi V. Dang,et al.  c-Myc Target Genes Involved in Cell Growth, Apoptosis, and Metabolism , 1999, Molecular and Cellular Biology.

[47]  T. Hirano,et al.  STAT3 orchestrates contradictory signals in cytokine‐induced G1 to S cell‐cycle transition , 1998, The EMBO journal.

[48]  J. Hennessey,et al.  Increase in percutaneous muscle biopsy yield with a suction-enhancement technique. , 1997, Journal of applied physiology.

[49]  E. Korzus,et al.  The Mitogen-activated Protein Kinase and JAK-STAT Signaling Pathways Are Required for an Oncostatin M-responsive Element-mediated Activation of Matrix Metalloproteinase 1 Gene Expression* , 1997, The Journal of Biological Chemistry.

[50]  M. Tarnopolsky,et al.  Changes in human skeletal muscle ultrastructure and force production after acute resistance exercise. , 1995, Journal of applied physiology.

[51]  D. J. Parry,et al.  Satellite cell activity is required for hypertrophy of overloaded adult rat muscle , 1994, Muscle & nerve.

[52]  E. Schultz,et al.  Satellite cell behavior during skeletal muscle growth and regeneration. , 1989, Medicine and science in sports and exercise.

[53]  D. Wessel,et al.  A method for the quantitative recovery of protein in dilute solution in the presence of detergents and lipids. , 1984, Analytical biochemistry.

[54]  A. Mauro.  SATELLITE CELL OF SKELETAL MUSCLE FIBERS , 1961, The Journal of biophysical and biochemical cytology.

[55]  S. Karnik,et al.  Role of nuclear unphosphorylated STAT3 in angiotensin II type 1 receptor-induced cardiac hypertrophy. , 2010, Cardiovascular research.

[56]  N. Mukaida,et al.  Cardiovascular , Pulmonary and Renal Pathology The Absence of Interleukin-6 Enhanced Arsenite-Induced Renal Injury by Promoting Autophagy of Tubular Epithelial Cells with Aberrant Extracellular Signal-Regulated Kinase Activation , 2010 .

[57]  P. Muñoz-Cánoves,et al.  Interleukin-6 is an essential regulator of satellite cell-mediated skeletal muscle hypertrophy. , 2008, Cell metabolism.

[58]  C. Fischer.  Interleukin-6 in acute exercise and training: what is the biological relevance? , 2006, Exercise immunology review.

[59]  F. López‐Soriano,et al.  Tumor necrosis factor-alpha exerts interleukin-6-dependent and -independent effects on cultured skeletal muscle cells. , 2002, Biochimica et biophysica acta.

[60]  T. Ogihara,et al.  Interleukin-6 stimulates c-myc expression and proliferation of cultured vascular smooth muscle cells. , 1990, Biochemistry international.

[61]  J. Bergstrom.  Percutaneous needle biopsy of skeletal muscle in physiological and clinical research. , 1975, Scandinavian journal of clinical and laboratory investigation.