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Related Experiment Videos

Decrease of intracellular ATP content downregulated UCP2 expression in mouse hepatocytes.

Gang Cheng1, Carmen C Polito, Julia K Haines

  • 1Division of Transplant Surgery, Department of Surgery, Medical University of South Carolina, Charleston 29425, USA.

Biochemical and Biophysical Research Communications
|August 14, 2003
PubMed
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Decreased cellular ATP levels, induced by FCCP and 2-deoxy-D-glucose, reduce mitochondrial uncoupling protein 2 (UCP2) mRNA expression in liver cells. ATP and UCP2 levels recovered upon removal of the inhibitors.

Area of Science:

  • Cellular biology
  • Mitochondrial function
  • Energy metabolism

Background:

  • Mitochondrial uncoupling protein 2 (UCP2) regulates energy metabolism and is upregulated in steatotic livers, increasing vulnerability to ischemic damage.
  • Previous research indicated increased UCP2 expression in steatotic livers, leading to diminished hepatic ATP and susceptibility to ischemic injury.

Purpose of the Study:

  • To investigate the effect of decreased intracellular ATP levels on UCP2 expression in a murine hepatocyte cell line (HEP6-16).
  • To mimic an ischemic liver state in vitro using ATP production inhibitors.

Main Methods:

  • Utilized Carbonyl cyanide p-trifluoromethoxyphenylhydrazone (FCCP), an oxidative phosphorylation uncoupler, to decrease intracellular ATP.
  • Combined FCCP with 2-deoxy-D-glucose, a glycolysis inhibitor, to potentiate ATP reduction.

Related Experiment Videos

  • Monitored UCP2 mRNA expression and intracellular ATP levels in HEP6-16 cells.
  • Assessed mitochondrial membrane potential changes in response to FCCP treatment.

    • Main Results:

    • FCCP dose- and time-dependently decreased intracellular ATP levels in HEP6-16 cells.
    • 2-deoxy-D-glucose significantly potentiated FCCP's inhibitory effect on ATP.
    • Decreased ATP levels correlated with reduced UCP2 mRNA expression.
    • ATP and UCP2 mRNA levels normalized within hours after inhibitor removal.
    • Mitochondrial membrane potential was only dissipated at high FCCP concentrations (80 microM), suggesting UCP2 downregulation was independent of this parameter.

    Conclusions:

    • In vitro reduction of intracellular ATP stores by FCCP and 2-deoxy-D-glucose leads to decreased UCP2 mRNA expression in hepatocytes.
    • The observed downregulation of UCP2 is not directly related to changes in mitochondrial membrane potential.
    • These findings in vitro support in vivo observations related to ischemia/reperfusion injury in the liver.