AMP-activated protein kinase-independent inhibition of hepatic mitochondrial oxidative phosphorylation by AICA riboside
Résumé
5-Aminoimidazole-4-carboxamide-1-β-D-ribofuranoside (AICA riboside) has been extensively used in cells to activate the AMP-activated protein kinase (AMPK), a metabolic sensor involved in cell energy homeostasis. In this work, we studied its effects on mitochondrial oxidative phosphorylation. AICA riboside was found to dose-dependently inhibit the oligomycin-sensitive oxygen consumption rate (JO 2}) of isolated rat hepatocytes. A decrease in inorganic phosphate (Pi), ATP, AMP and total adenine nucleotide contents was also observed with [AICA riboside] > 0.1 mM. Interestingly, in hepatocytes from mice lacking both α1 and α2 AMPK catalytic subunits, basal JO 2} and expression of several mitochondrial proteins were significantly reduced compared to wild-type mice, suggesting that mitochondrial biogenesis was perturbed. However, inhibition of JO 2} by AICA riboside was still present in the mutant mice and thus was clearly not mediated by AMPK. In permeabilized hepatocytes, this inhibition was no longer evidenced, suggesting that it could be due to intracellular accumulation of Z nucleotides and/or loss of adenine nucleotides and Pi. ZMP did indeed inhibit respiration in isolated rat mitochondria through a direct effect on the respiratory-chain complex 1. In addition, inhibition of JO 2} by AICA riboside was also potentiated in cells incubated with fructose to deplete adenine nucleotides and Pi. We conclude that AICA riboside inhibits cellular respiration by an AMPK-independent mechanism that likely results from the combined intracellular Pi depletion and ZMP accumulation. Our data also demonstrate that the cellular effects of AICA riboside are not necessarily caused by AMPK activation and thus that their interpretation should be taken with caution.
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