Effect of caffeine, caffeic acid and their various combinations on enzymes of cholinergic, monoaminergic and purinergic systems critical to neurodegeneration in rat brain-In vitro

Neurotoxicology. 2017 Sep;62:6-13. doi: 10.1016/j.neuro.2017.04.008. Epub 2017 Apr 29.


Caffeine and caffeic acid are two bioactive compounds that are present in plant foods and are major constituent of coffee, cocoa, tea, cola drinks and chocolate. Although not structurally related, caffeine and caffeic acid has been reported to elicit neuroprotective properties. However, their different proportional distribution in food sources and possible effect of such interactions are not often taken into consideration. Therefore, in this study, we investigated the effect of caffeine, caffeic acid and their various combinations on activities of some enzymes [acetylcholinesterase (AChE), monoamine oxidase (MAO) ecto-nucleoside triphosphate diphosphohydrolase (E-NTPase), ecto-51-nucleotidase (E-NTDase) and Na+/K+ ATPase relevant to neurodegeneration in vitro in rat brain. The stock concentration of caffeine and caffiec acid and their various proportional combinations were prepared and their interactions with the activities of these enzymes were assessed (in vitro) in different brain structures. The Fe2+ and Cu2+ chelating abilities of the samples were also investigated. The results revealed that caffeine, caffeic acid and their various combinations exhibited inhibitory effect on activities of AChE, MAO, E-NTPase and E-NTDase, but stimulatory effect on Na+/K+ ATPase activity. The combinations also exhibited Fe2+ and Cu2+ chelating abilities. Considering the various combinations, a higher caffeine to caffeic acid ratio produced significantly highest enzyme modulatory effects; these were significantly lower to the effect of caffeine alone but significantly higher than the effect of caffeic acid alone. These findings may provide new insight into the effect of proportional combination of these bioactive compounds as obtained in many foods especially with respect to their neuroprotective effects.

Keywords: Acetylcholinesterase; Caffeic acid; Caffeine; Monoamine oxidase; Neuromodulation; Purinergic signalling.

MeSH terms

  • Acetylcholinesterase / metabolism*
  • Adenosine Triphosphatases / metabolism*
  • Animals
  • Brain / drug effects*
  • Caffeic Acids / pharmacology*
  • Caffeine / pharmacology*
  • Central Nervous System Stimulants / pharmacokinetics*
  • Copper / metabolism
  • Dose-Response Relationship, Drug
  • Drug Combinations
  • Iron / metabolism
  • Monoamine Oxidase / metabolism*
  • Rats
  • Rats, Wistar
  • Spectrophotometry


  • Caffeic Acids
  • Central Nervous System Stimulants
  • Drug Combinations
  • Caffeine
  • Copper
  • Iron
  • Monoamine Oxidase
  • Acetylcholinesterase
  • Adenosine Triphosphatases
  • caffeic acid