Effects of WY-14,643 on the phosphorylation and activation of AMP-dependent protein kinase

Arch Biochem Biophys. 2009 May 1;485(1):10-5. doi: 10.1016/j.abb.2009.02.006. Epub 2009 Feb 21.


Background: AMP-dependent protein kinase (AMPK) and peroxisome proliferator-activated receptor (PPAR) alpha facilitate fatty acid oxidation. We have shown that treatment of hepatoma cells with ethanol or feeding ethanol-containing diets to mice inhibited both PPARalpha and AMPK activity. Importantly, WY-14,643 reversed the development of fatty liver in alcohol-fed mice. Whether WY-14,643, a PPARalpha agonist, has any effects on AMPK is not known. The aim of this study was to investigate the effect of WY-14,643 on AMPK activity.

Methods: The effect of WY-14,643 on AMPK phosphorylation and activity were examined in rat hepatoma cells (H4IIEC3). The effect of WY-14,643 on upstream kinases of AMPK, PKC-zeta/LKB1, intracellular AMP:ATP ratio, oxidative stress, and AMPK gene expression were studied.

Results: Treatment of the H4IIEC3 cells with WY-14,643 for 24h led to 60% increase in the phosphorylation of AMPK. The effect of WY-14,643 on AMPK phosphorylation is PKC-zeta/LKB1 independent. WY-14,643 did not alter the levels of intracellular AMP:ATP ratio and it did not increase the levels of reactive oxygen species at 24-h of treatment. WY-14,643-induced AMPK alpha subunit expression by 2- to 2.5-fold, but there was no change in AMPKalpha subunit protein at 24h. The effect of WY-14,643 on AMPK phosphorylation did not altered by the presence of an NADPH oxidase inhibitor.

Conclusions: WY-14,643 induced AMPKalpha subunit phosphorylation and the activity of the enzyme. This was associated with induction of AMPKalpha1 and alpha2 mRNA, but the mechanism for this activation is uncertain.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • AMP-Activated Protein Kinases / genetics
  • AMP-Activated Protein Kinases / metabolism*
  • Adenosine Monophosphate / metabolism
  • Adenosine Triphosphate / metabolism
  • Cell Line
  • Energy Metabolism / drug effects
  • Enzyme Activation / drug effects
  • Gene Expression Regulation, Enzymologic / drug effects
  • PPAR alpha / metabolism
  • Phosphorylation / drug effects
  • Protein Kinase C / metabolism
  • Pyrimidines / pharmacology*
  • Reactive Oxygen Species / metabolism


  • PPAR alpha
  • Pyrimidines
  • Reactive Oxygen Species
  • Adenosine Monophosphate
  • pirinixic acid
  • Adenosine Triphosphate
  • protein kinase C zeta
  • Protein Kinase C
  • AMP-Activated Protein Kinases