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Diabetologia
Published in: Diabetologia 12/2011

Open Access 01-12-2011 | Article

Metformin activates AMP-activated protein kinase in primary human hepatocytes by decreasing cellular energy status

Authors: X. Stephenne, M. Foretz, N. Taleux, G. C. van der Zon, E. Sokal, L. Hue, B. Viollet, B. Guigas

Published in: Diabetologia | Issue 12/2011

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Abstract

Aim/hypothesis

The glucose-lowering drug metformin has been shown to activate hepatic AMP-activated protein kinase (AMPK), a master kinase regulating cellular energy homeostasis. However, the underlying mechanisms remain controversial and have never been investigated in primary human hepatocytes.

Methods

Hepatocytes isolated from rat, mouse and human livers were treated with various concentrations of metformin. Isoform-specific AMPKα abundance and activity, as well as intracellular adenine nucleotide levels and mitochondrial oxygen consumption rates were determined at different time points.

Results

Metformin dose- and time-dependently increased AMPK activity in rat and human hepatocytes, an effect associated with a significant rise in cellular AMP:ATP ratio. Surprisingly, we found that AMPKα2 activity was undetectable in human compared with rat hepatocytes, while AMPKα1 activities were comparable. Accordingly, metformin only increased AMPKα1 activity in human hepatocytes, although both AMPKα isoforms were activated in rat hepatocytes. Analysis of mRNA expression and protein levels confirmed that only AMPKα1 is present in human hepatocytes; it also showed that the distribution of β and γ regulatory subunits differed between species. Finally, we demonstrated that the increase in AMP:ATP ratio in hepatocytes from liver-specific Ampkα1/2 (also known as Prkaa1/2) knockout mice and humans is due to a similar and specific inhibition of the mitochondrial respiratory-chain complex 1 by metformin.

Conclusions/interpretation

Activation of hepatic AMPK by metformin results from a decrease in cellular energy status owing to metformin’s AMPK-independent inhibition of the mitochondrial respiratory-chain complex 1. The unique profile of AMPK subunits found in human hepatocytes should be considered when developing new pharmacological agents to target the kinase.
Appendix
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Metadata
Title
Metformin activates AMP-activated protein kinase in primary human hepatocytes by decreasing cellular energy status
Authors
X. Stephenne
M. Foretz
N. Taleux
G. C. van der Zon
E. Sokal
L. Hue
B. Viollet
B. Guigas
Publication date
01-12-2011
Publisher
Springer-Verlag
Published in
Diabetologia / Issue 12/2011
Print ISSN: 0012-186X
Electronic ISSN: 1432-0428
DOI
https://doi.org/10.1007/s00125-011-2311-5

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