Journal of Physiology and Biochemistry

, Volume 74, Issue 1, pp 35–45 | Cite as

Metabolic effects of physiological levels of caffeine in myotubes

  • Jamie K. Schnuck
  • Lacey M. Gould
  • Hailey A. Parry
  • Michele A. Johnson
  • Nicholas P. Gannon
  • Kyle L. Sunderland
  • Roger A. VaughanEmail author
Original Article


Caffeine has been shown to stimulate multiple major regulators of cell energetics including AMP-activated protein kinase (AMPK) and Ca2+/calmodulin-dependent protein kinase II (CaMKII). Additionally, caffeine induces peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1α) and mitochondrial biogenesis. While caffeine enhances oxidative metabolism, experimental concentrations often exceed physiologically attainable concentrations through diet. This work measured the effects of low-level caffeine on cellular metabolism and gene expression in myotubes, as well as the dependence of caffeine’s effects on the nuclear receptor peroxisome proliferator-activated receptor beta/delta (PPARβ/δ). C2C12 myotubes were treated with various doses of caffeine for up to 24 h. Gene and protein expression were measured via qRT-PCR and Western blot, respectively. Cellular metabolism was determined via oxygen consumption and extracellular acidification rate. Caffeine significantly induced regulators of mitochondrial biogenesis and oxidative metabolism. Mitochondrial staining was suppressed in PPARβ/δ-inhibited cells which was rescued by concurrent caffeine treatment. Caffeine-treated cells also displayed elevated peak oxidative metabolism which was partially abolished following PPARβ/δ inhibition. Similar to past observations, glucose uptake and GLUT4 content were elevated in caffeine-treated cells, however, glycolytic metabolism was unaltered following caffeine treatment. Physiological levels of caffeine appear to enhance cell metabolism through mechanisms partially dependent on PPARβ/δ.


PGC-1α PPARβ/δ Skeletal muscle Mitochondrial biogenesis GLUT4 



Fatty acyl-coA dehydrogenase


Acetyl-coA carboxylase


AMP-activated protein kinase


Ca2+/calmodulin-dependent protein kinases I/II


Carbohydrate-responsive element-binding protein


cAMP response element-binding protein


Citrate synthase


Estrogen-related receptor alpha


Fatty acid synthase


Forkhead box protein O1


Glucose transporter 4


Lactate dehydrogenase a


Lactate dehydrogenase b


Myocyte enhancer factor 2


Nuclear respiratory factor 1


Peroxisome proliferator-activated receptor gamma coactivator 1-alpha


Peroxisome proliferator-activated receptor alpha


Peroxisome proliferator-activated receptor beta/delta


Peroxisome proliferator-activated receptor gamma


Stearoyl-CoA desaturase


Sterol receptor element-binding protein-1c


NAD+-dependent deacetylase sirtuin 3


Mitochondrial transcription factor A


Uncoupling protein



We would like to thank the Department of Physical Therapy (Congdon School of Health Sciences) and the School of Arts and Sciences for the use of shared lab space and equipment.

Author contribution

All authors read and approved the final manuscript. JKS, LMG, HAP, MAJ, and NPG assisted with experiments and manuscript preparation. KLS assisted with experimental design and manuscript preparation. RAV performed and oversaw all experiments, statistical analyses, manuscript preparation, and designed/conceived the experiments.

Funding information

Support for this work was provided by the Department of Exercise Science within the Congdon School of Health Sciences.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

13105_2017_601_MOESM1_ESM.jpeg (40 kb)
Figure S1 Cell viability. WST-1 assay time trial of cells treated with or without caffeine at 100 μM for 24 hours (left) and corresponding area-under-the-curve (right). Notes: Viability was performed using 10 replicates per treatment and analyzed using two-way ANOVA with Bonferroni’s correction for multiple comparisons. (JPEG 40 kb)


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Copyright information

© University of Navarra 2017

Authors and Affiliations

  • Jamie K. Schnuck
    • 1
    • 2
  • Lacey M. Gould
    • 1
  • Hailey A. Parry
    • 1
    • 3
  • Michele A. Johnson
    • 1
  • Nicholas P. Gannon
    • 2
  • Kyle L. Sunderland
    • 1
  • Roger A. Vaughan
    • 1
    Email author
  1. 1.Department of Exercise ScienceHigh Point UniversityHigh PointUSA
  2. 2.School of MedicineMedical College of WisconsinMilwaukeeUSA
  3. 3.School of KinesiologyAuburn UniversityAuburnUSA

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