Metformin reduces hepatic expression of SIRT3, the mitochondrial deacetylase controlling energy metabolism

• Published in: PloS one 2012-11, Vol.7 (11), p.e49863
• Main Authors: Buler, Marcin, Aatsinki, Sanna-Mari, Izzi, Valerio, Hakkola, Jukka
• Format: Article
• Language: English
• Subjects: Acetylation; Adenosine Triphosphate - metabolism; AMP-activated protein kinase; Analysis of Variance; Animals; Antidiabetics; ATP; Biology; Biosynthesis; Cell cycle; Cellular biology; Cyclic adenosine monophosphate; Cyclic AMP; Deoxyribonucleic acid; Diabetes; Diabetes therapy; DNA; DNA Primers - genetics; DNA, Mitochondrial - metabolism; Electron transport; Electron transport chain; Energy; Energy metabolism; Energy Metabolism - drug effects; ERRalpha Estrogen-Related Receptor; Estrogens; Fatty acids; Female; Flow Cytometry; Gene expression; Gene Expression Regulation - drug effects; Glucagon; Glucose; Hep G2 Cells; Hepatocytes; Hepatocytes - metabolism; Homeostasis; Humans; Immunoblotting; Kinases; Laboratory animals; Liver; Medicine; Metabolism; Metformin; Metformin - pharmacology; Mice; Microscopy, Fluorescence; Mitochondria; Mitochondrial DNA; Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1-alpha; Pharmacology; Phosphorylation; Protein kinases; Proteins; Real-Time Polymerase Chain Reaction; Receptors, Estrogen - metabolism; Regulation; RNA; RNA, Small Interfering - genetics; Rodents; siRNA; Sirtuin 3 - metabolism; Toxicology; Trans-Activators - metabolism; Transcription Factors; Type 2 diabetes
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0049863

Metformin inhibits ATP production in mitochondria and this may be involved in the anti-hyperglycemic effects of the drug. Sirtuin 3 (SIRT3) is a mitochondrial protein deacetylase that regulates the function of the electron transport chain and maintains basal ATP yield. We hypothesized that metformin treatment could diminish mitochondrial ATP production through downregulation of SIRT3 expression. Glucagon and cAMP induced SIRT3 mRNA in mouse primary hepatocytes. Metformin prevented SIRT3 induction by glucagon. Moreover, metformin downregulated constitutive expression of SIRT3 in primary hepatocytes and in the liver in vivo. Estrogen related receptor alpha (ERRα) mediates regulation of Sirt3 gene by peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1α). ERRα mRNA expression was regulated in a similar manner as SIRT3 mRNA by glucagon, cAMP and metformin. However, a higher metformin concentration was required for downregulation of ERRα than SIRT3. ERRα siRNA attenuated PGC-1α mediated induction of SIRT3, but did not affect constitutive expression. Overexpression of the constitutively active form of AMP-activated protein kinase (AMPK) induced SIRT3 mRNA, indicating that the SIRT3 downregulation by metformin is not mediated by AMPK. Metformin reduced the hepatocyte ATP level. This effect was partially counteracted by SIRT3 overexpression. Furthermore, metformin decreased mitochondrial SIRT3 protein levels and this was associated with enhanced acetylation of several mitochondrial proteins. However, metformin increased mitochondrial mass in hepatocytes. Altogether, our results indicate that metformin attenuates mitochondrial expression of SIRT3 and suggest that this mechanism is involved in regulation of energy metabolism by metformin in the liver and may contribute to the therapeutic action of metformin.