Cited by 287Metabolism, Diabetes, and CancerBiochemical and Molecular ResearchPeroxisome Proliferator-Activated Receptors

Metformin Activates AMP-Activated Protein Kinase in Primary Human Hepatocytes by Decreasing Cellular Energy Status

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Stephenne, X., Foretz, M., Taleux, N., van der Zon, G. C., Sokal, E., Hue, L., Viollet, B., Guigas, B.·2011·Diabetologia

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.

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Authors
Stephenne, X., Foretz, M., Taleux, N., van der Zon, G. C., Sokal, E., Hue, L., Viollet, B., & Guigas, B.
Affiliations
  • Centre National de la Recherche Scientifique (FR)
  • Inserm (FR)
  • Cliniques Universitaires Saint-Luc (BE)
  • de Duve Institute (BE)
  • Leiden University Medical Center (NL)
Corresponding author
Guigas, B.
Venue
Diabetologia
Published
2011-09-23
Volume
54
Issue
12
Pages
3101-3110
ISSN
  • 0012-186X (ISSN-L)
  • 1432-0428
OpenAlex
W2042871909
Cited by
287
References
52
Citations by year
2026: 14 · 2025: 12 · 2024: 20 · 2023: 10 · 2022: 22 · 2021: 14 · 2020: 24 · 2019: 30 · 2018: 25 · 2017: 26 · 2016: 19 · 2015: 16 · 2014: 22 · 2013: 19 · 2012: 12
FWCI
6.97
Type
journal article
Language
en
Open access
Licence
cc-by-nc
Locations
  • Diabetologia (publishedVersion, cc-by-nc)
  • Europe PMC (PubMed Central) (publishedVersion, cc-by-nc)
  • PubMed Central (publishedVersion, cc-by-nc)
  • PubMed (publishedVersion)
  • Data Archiving and Networked Services (DANS) (submittedVersion)
  • Leiden Repository (Leiden University) (submittedVersion)
  • DIAL (Catholic University of Leuven) (submittedVersion)
Funders
  • European Commission (LSHMCT-2004-005272)
  • Agence Nationale de la Recherche (ANR-10-BLAN-1123, ANR-10)
  • Fonds De La Recherche Scientifique - FNRS
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Metabolism, Diabetes, and Cancer
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Biochemical and Molecular Research
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Peroxisome Proliferator-Activated Receptors
Keywords
metformin, AMPK, AMPKα1, AMPKα2, human hepatocytes, cellular energy status, AMP/ATP ratio, mitochondrial complex I inhibition
MeSH terms
Adenine Nucleotides, Animals, Cells, Cultured, Humans, Hypoglycemic Agents, Isoenzymes, Male, Metformin, Mitochondria, Liver, Oxygen Consumption, Mice, Knockout, Hepatocytes, Mice, Rats, AMP-Activated Protein Kinases

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