Effects of long-term treatment with dietary theobromine on rat skeletal muscles

Mol Biol Rep. 2022 May;49(5):3965-3973. doi: 10.1007/s11033-022-07248-w. Epub 2022 Feb 28.


Background: Plastic changes of skeletal muscles, such as hypertrophy and atrophy, are dependent on physiological activities and regulated by a variety of signaling pathways, including cyclic adenosine monophosphate (cAMP) pathway. The cAMP inducing agents, such as the β2-adrenergic agonist clenbuterol, are known to induce muscle hypertrophy, and has been reported to induce slow-to-fast transitions in rat soleus muscle. Theobromine, one of the active components of cacao, functions as an inhibitor of phosphodiesterase and increases cAMP. This study hypothesized that theobromine, like clenbuterol, can induce muscle hypertrophy and influence contractile properties.

Methods and results: Male Wistar rats were fed a normal diet or a diet containing 0.05% theobromine for 20 weeks. Using biochemical, anatomical, and physiological techniques, effects of dietary theobromine on skeletal muscles (soleus, extensor digitorum longus, plantaris, and gastrocnemius) were examined. There were no significant differences in body weight, serum levels of proteins and lipids, muscle weights, dry/wet ratio of muscle weights, mitochondrial oxidation enzyme activity of muscles, isometric contractile properties of muscles, and muscle fatigue between control and theobromine-fed rats. Quantitative analysis of mRNA, however, revealed upregulation of myosin heavy chain 2x and myogenic differentiation 1, as previously reported in clenbuterol-treated muscles.

Conclusion: The long-term theobromine (0.05%) diet in rats had no effect in inducing muscle hypertrophy and in changing contractile properties, although it had some similar effects of clenbuterol on muscle gene expression.

Keywords: Contractile property; Hypertrophy; Myogenic differentiation 1; Myosin heavy chain 2x; Rat skeletal muscle; Theobromine.

MeSH terms

  • Adrenergic beta-Agonists / metabolism
  • Animals
  • Clenbuterol* / analysis
  • Clenbuterol* / metabolism
  • Clenbuterol* / pharmacology
  • Diet
  • Hypertrophy
  • Male
  • Muscle, Skeletal / metabolism
  • Rats
  • Rats, Wistar
  • Theobromine / analysis
  • Theobromine / metabolism
  • Theobromine / pharmacology


  • Adrenergic beta-Agonists
  • Theobromine
  • Clenbuterol