Metabolic effects of physiological levels of caffeine in myotubes

J Physiol Biochem. 2018 Feb;74(1):35-45. doi: 10.1007/s13105-017-0601-1. Epub 2017 Dec 3.


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.

Keywords: GLUT4; Mitochondrial biogenesis; PGC-1α; PPARβ/δ; Skeletal muscle.

MeSH terms

  • Animals
  • Benzamides / pharmacology
  • Biological Assay
  • Caffeine / metabolism*
  • Cell Line
  • Coculture Techniques
  • Gene Expression Regulation* / drug effects
  • Hydrogen-Ion Concentration
  • Lipid Metabolism / drug effects
  • Mice
  • Mitochondria, Muscle / drug effects
  • Mitochondria, Muscle / enzymology
  • Mitochondria, Muscle / metabolism*
  • Mitochondrial Dynamics / drug effects
  • Muscle Fibers, Skeletal / drug effects
  • Muscle Fibers, Skeletal / metabolism*
  • Muscle Proteins / agonists
  • Muscle Proteins / antagonists & inhibitors
  • Muscle Proteins / genetics
  • Muscle Proteins / metabolism*
  • Organelle Biogenesis
  • Osmolar Concentration
  • Oxidative Phosphorylation / drug effects
  • Oxygen Consumption / drug effects
  • PPAR delta / agonists*
  • PPAR delta / antagonists & inhibitors
  • PPAR delta / metabolism
  • PPAR-beta / agonists*
  • PPAR-beta / antagonists & inhibitors
  • PPAR-beta / metabolism
  • Smegmamorpha
  • Spheroids, Cellular / drug effects
  • Spheroids, Cellular / metabolism
  • Sulfones / pharmacology


  • 4-chloro-N-(2-((5-trifluoromethyl-2-pyridyl)sulfonyl)ethyl)benzamide
  • Benzamides
  • Muscle Proteins
  • PPAR delta
  • PPAR-beta
  • Sulfones
  • Caffeine