Resveratrol Ameliorates Aging-Related Metabolic Phenotypes by Inhibiting cAMP Phosphodiesterases

Resveratrol, a polyphenol in red wine, has been reported as a calorie restriction mimetic with potential antiaging and antidiabetogenic properties. It is widely consumed as a nutritional supplement, but its mechanism of action remains a mystery. Here, we report that the metabolic effects of resveratrol result from competitive inhibition of cAMP-degrading phosphodiesterases, leading to elevated cAMP levels. The resulting activation of Epac1, a cAMP effector protein, increases intracellular Ca(2+) levels and activates the CamKKβ-AMPK pathway via phospholipase C and the ryanodine receptor Ca(2+)-release channel. As a consequence, resveratrol increases NAD(+) and the activity of Sirt1. Inhibiting PDE4 with rolipram reproduces all of the metabolic benefits of resveratrol, including prevention of diet-induced obesity and an increase in mitochondrial function, physical stamina, and glucose tolerance in mice. Therefore, administration of PDE4 inhibitors may also protect against and ameliorate the symptoms of metabolic diseases associated with aging.

[1]  H. Rehmann.  Characterization of the activation of the Rap-specific exchange factor Epac by cyclic nucleotides. , 2006, Methods in enzymology.

[2]  D. Schomburg,et al.  Ca 2+ /calmodulin-dependent protein kinase , 1997 .

[3]  P. Defossez,et al.  Requirement of NAD and SIR2 for life-span extension by calorie restriction in Saccharomyces cerevisiae. , 2000, Science.

[4]  M. vanTriest,et al.  Measurement of GTP-bound Ras-like GTPases by activation-specific probes. , 2001 .

[5]  M. Beal,et al.  Attenuation of MPTP neurotoxicity by rolipram, a specific inhibitor of phosphodiesterase IV , 2008, Experimental Neurology.

[6]  Timothy Hagen,et al.  Design of phosphodiesterase 4D (PDE4D) allosteric modulators for enhancing cognition with improved safety , 2010, Nature Biotechnology.

[7]  P. Pfluger,et al.  Sirt1 protects against high-fat diet-induced metabolic damage , 2008, Proceedings of the National Academy of Sciences.

[8]  Phuong Chung,et al.  Small molecule activators of sirtuins extend Saccharomyces cerevisiae lifespan , 2003, Nature.

[9]  Stephan E. Lehnart,et al.  Ca2+/Calmodulin-Dependent Protein Kinase II Phosphorylation Regulates the Cardiac Ryanodine Receptor , 2004, Circulation research.

[10]  P. Puigserver,et al.  AMPK regulates energy expenditure by modulating NAD+ metabolism and SIRT1 activity , 2009, Nature.

[11]  P. Puigserver,et al.  Resveratrol improves health and survival of mice on a high-calorie diet , 2006, Nature.

[12]  B. Kemp.  Faculty Opinions recommendation of Complexes between the LKB1 tumor suppressor, STRAD alpha/beta and MO25 alpha/beta are upstream kinases in the AMP-activated protein kinase cascade. , 2003 .

[13]  Jiandie D. Lin,et al.  Transcriptional co-activator PGC-1α drives the formation of slow-twitch muscle fibres , 2002, Nature.

[14]  J. Bos,et al.  Measurement of GTP-bound Ras-like GTPases by activation-specific probes. , 2001, Methods in enzymology.

[15]  F. Ross,et al.  Use of Cells Expressing γ Subunit Variants to Identify Diverse Mechanisms of AMPK Activation , 2010, Cell metabolism.

[16]  F. Mulero,et al.  Sirt1 improves healthy ageing and protects from metabolic syndrome-associated cancer. , 2010, Nature communications.

[17]  P. Puigserver,et al.  Metabolic control of muscle mitochondrial function and fatty acid oxidation through SIRT1/PGC‐1α , 2007, The EMBO journal.

[18]  J. Auwerx,et al.  Specific SIRT1 activation mimics low energy levels and protects against diet-induced metabolic disorders by enhancing fat oxidation. , 2008, Cell metabolism.

[19]  J. Chayvialle,et al.  Co-requirement of Cyclic AMP- and Calcium-dependent Protein Kinases for Transcriptional Activation of Cholecystokinin Gene by Protein Hydrolysates* , 2002, The Journal of Biological Chemistry.

[20]  B. Blaxall,et al.  Epac and Phospholipase Cϵ Regulate Ca2+ Release in the Heart by Activation of Protein Kinase Cϵ and Calcium-Calmodulin Kinase II* , 2009, Journal of Biological Chemistry.

[21]  Wilhelm Haas,et al.  Nutrient control of glucose homeostasis through a complex of PGC-1α and SIRT1 , 2005, Nature.

[22]  M. Tota,et al.  Crystal structure of human phosphodiesterase 3B: atomic basis for substrate and inhibitor specificity. , 2004, Biochemistry.

[23]  A. Wittinghofer,et al.  Epac is a Rap1 guanine-nucleotide-exchange factor directly activated by cyclic AMP , 1998, Nature.

[24]  P. Strålfors,et al.  Differential regulation of adipocyte PDE3B in distinct membrane compartments by insulin and the beta3-adrenergic receptor agonist CL316243: effects of caveolin-1 knockdown on formation/maintenance of macromolecular signalling complexes. , 2009, The Biochemical journal.

[25]  Alexander S Banks,et al.  SirT1 gain of function increases energy efficiency and prevents diabetes in mice. , 2008, Cell metabolism.

[26]  M. Smigel.  Purification of the catalyst of adenylate cyclase. , 1986, The Journal of biological chemistry.

[27]  H. Ke,et al.  Crystal structures of phosphodiesterases and implications on substrate specificity and inhibitor selectivity. , 2007, Current topics in medicinal chemistry.

[28]  Brian C. Smith,et al.  Mechanism of Human SIRT1 Activation by Resveratrol* , 2005, Journal of Biological Chemistry.

[29]  Amy V. Lynch,et al.  Small molecule activators of SIRT1 as therapeutics for the treatment of type 2 diabetes , 2007, Nature.

[30]  A M Graybiel,et al.  A family of cAMP-binding proteins that directly activate Rap1. , 1998, Science.

[31]  C. Rondinone,et al.  Phosphorylation of PDE3B by Phosphatidylinositol 3-Kinase Associated with the Insulin Receptor* , 2000, The Journal of Biological Chemistry.

[32]  K. Jakobs,et al.  A new phospholipase-C–calcium signalling pathway mediated by cyclic AMP and a Rap GTPase , 2001, Nature Cell Biology.

[33]  J. Simon,et al.  AMP-activated Protein Kinase Signaling Activation by Resveratrol Modulates Amyloid-β Peptide Metabolism* , 2010, The Journal of Biological Chemistry.

[34]  B. Sümegi,et al.  Resveratrol improves insulin sensitivity, reduces oxidative stress and activates the Akt pathway in type 2 diabetic patients. , 2011, The British journal of nutrition.

[35]  F. Lezoualc’h,et al.  The cAMP binding protein Epac modulates Ca2+ sparks by a Ca2+/calmodulin kinase signalling pathway in rat cardiac myocytes , 2007, The Journal of physiology.

[36]  Jiandie D. Lin,et al.  Bioenergetic analysis of peroxisome proliferator-activated receptor gamma coactivators 1alpha and 1beta (PGC-1alpha and PGC-1beta) in muscle cells. , 2003, The Journal of biological chemistry.

[37]  A. Edelman,et al.  Calmodulin-dependent protein kinase kinase-beta is an alternative upstream kinase for AMP-activated protein kinase. , 2005, Cell metabolism.

[38]  S. Fields,et al.  Substrate-specific Activation of Sirtuins by Resveratrol* , 2005, Journal of Biological Chemistry.

[39]  O. Arancio,et al.  Reversal of long-term dendritic spine alterations in Alzheimer disease models , 2009, Proceedings of the National Academy of Sciences.

[40]  C M McCay,et al.  The effect of retarded growth upon the length of life span and upon the ultimate body size. 1935. , 1935, Nutrition.

[41]  L. Guarente.  Sirtuins as potential targets for metabolic syndrome , 2006, Nature.

[42]  R. Ghidoni,et al.  Resveratrol as an anticancer nutrient: molecular basis, open questions and promises. , 2005, The Journal of nutritional biochemistry.

[43]  J. Wood,et al.  Sirtuin activators mimic caloric restriction and delay ageing in metazoans , 2004, Nature.

[44]  David Carling,et al.  A common bicyclic protein kinase cascade inactivates the regulatory enzymes of fatty acid and cholesterol biosynthesis , 1987, FEBS letters.

[45]  P. Puigserver,et al.  Resveratrol Improves Mitochondrial Function and Protects against Metabolic Disease by Activating SIRT1 and PGC-1α , 2006, Cell.

[46]  P. Puigserver,et al.  Concurrent regulation of AMP-activated protein kinase and SIRT1 in mammalian cells. , 2009, Biochemical and biophysical research communications.

[47]  David Carling,et al.  Supplemental Data LKB 1 Is the Upstream Kinase in the AMP-Activated Protein Kinase Cascade , 2003 .

[48]  R. Heath,et al.  Ca2+/calmodulin-dependent protein kinase kinase-beta acts upstream of AMP-activated protein kinase in mammalian cells. , 2005, Cell metabolism.

[49]  S. Bastianetto,et al.  Faculty Opinions recommendation of SIRT1 suppresses beta-amyloid production by activating the alpha-secretase gene ADAM10. , 2010 .

[50]  F. Orallo,et al.  Effects of trans- and cis-resveratrol on Ca2+ handling in A7r5 vascular myocytes. , 2007, European journal of pharmacology.

[51]  Jiandie D. Lin,et al.  Bioenergetic Analysis of Peroxisome Proliferator-activated Receptor γ Coactivators 1α and 1β (PGC-1α and PGC-1β) in Muscle Cells* , 2003, Journal of Biological Chemistry.

[52]  A. Means,et al.  The Ca2+/Calmodulin-dependent Protein Kinase Kinases Are AMP-activated Protein Kinase Kinases* , 2005, Journal of Biological Chemistry.

[53]  B. Halliwell,et al.  Evidence for a Trade‐Off between Survival and Fitness Caused by Resveratrol Treatment of Caenorhabditis elegans , 2007, Annals of the New York Academy of Sciences.

[54]  Shu-Chen Lu,et al.  Resveratrol is Not a Direct Activator of SIRT1 Enzyme Activity , 2009, Chemical biology & drug design.

[55]  Leonid Peshkin,et al.  Resveratrol delays age-related deterioration and mimics transcriptional aspects of dietary restriction without extending life span. , 2008, Cell metabolism.

[56]  Joseph A. Baur,et al.  Therapeutic potential of resveratrol: the in vivo evidence , 2006, Nature Reviews Drug Discovery.

[57]  F. Ross,et al.  Use of Cells Expressing gamma Subunit Variants to Identify Diverse Mechanisms of AMPK Activation , 2010 .

[58]  Venkataraman Thanabal,et al.  SRT1720, SRT2183, SRT1460, and Resveratrol Are Not Direct Activators of SIRT1♦ , 2010, The Journal of Biological Chemistry.

[59]  A. El-Mowafy,et al.  Resveratrol activates adenylyl-cyclase in human breast cancer cells: a novel, estrogen receptor-independent cytostatic mechanism. , 2003, Carcinogenesis.

[60]  J. Ha,et al.  Resveratrol stimulates glucose transport in C2C12 myotubes by activating AMP-activated protein kinase , 2007, Experimental & Molecular Medicine.

[61]  L. Guarente,et al.  RETRACTED: SIRT1 Suppresses β-Amyloid Production by Activating the α-Secretase Gene ADAM10 , 2010, Cell.

[62]  P. Puigserver,et al.  A Cold-Inducible Coactivator of Nuclear Receptors Linked to Adaptive Thermogenesis , 1998, Cell.

[63]  David S. Goodsell,et al.  A semiempirical free energy force field with charge‐based desolvation , 2007, J. Comput. Chem..

[64]  R. Taussig,et al.  Expression and purification of recombinant adenylyl cyclases in Sf9 cells. , 1994, Methods in enzymology.

[65]  E. Hoffman,et al.  Glucose restriction inhibits skeletal myoblast differentiation by activating SIRT1 through AMPK-mediated regulation of Nampt. , 2008, Developmental cell.

[66]  Jérôme Boudeau,et al.  Complexes between the LKB1 tumor suppressor, STRADα/β and MO25α/β are upstream kinases in the AMP-activated protein kinase cascade , 2003, Journal of biology.

[67]  B. Viollet,et al.  AMP-Activated Protein Kinase–Deficient Mice Are Resistant to the Metabolic Effects of Resveratrol , 2009, Diabetes.

[68]  J. Auwerx,et al.  Interdependence of AMPK and SIRT1 for metabolic adaptation to fasting and exercise in skeletal muscle. , 2010, Cell metabolism.

[69]  F. Méchali,et al.  RAP1A GTP/GDP cycles determine the intracellular location of the late endocytic compartments and contribute to myogenic differentiation. , 1999, Experimental cell research.

[70]  David Carling,et al.  Structure of Mammalian AMPK and its regulation by ADP , 2011, Nature.

[71]  J. Milbrandt,et al.  Resveratrol stimulates AMP kinase activity in neurons , 2007, Proceedings of the National Academy of Sciences.

[72]  Jiandie D. Lin,et al.  Transcriptional co-activator PGC-1 alpha drives the formation of slow-twitch muscle fibres. , 2002, Nature.

[73]  Andrew D. Steele,et al.  SIRT1 transgenic mice show phenotypes resembling calorie restriction , 2007, Aging cell.

[74]  Matt Kaeberlein,et al.  Sir2-Independent Life Span Extension by Calorie Restriction in Yeast , 2004, PLoS biology.

[75]  Huan Wang,et al.  Epac-mediated Activation of Phospholipase Cϵ Plays a Critical Role in β-Adrenergic Receptor-dependent Enhancement of Ca2+ Mobilization in Cardiac Myocytes* , 2007, Journal of Biological Chemistry.

[76]  J. Crowell,et al.  Resveratrol-associated renal toxicity. , 2004, Toxicological sciences : an official journal of the Society of Toxicology.

[77]  L. Recant,et al.  The effect of starvation on tissue adenosine 3'-5' monophosphate levels. , 1973, Biochemical and biophysical research communications.

[78]  V. Manganiello,et al.  An effect of insulin on cyclic adenosine 3':5'-monophosphate phosphodiesterase activity in fat cells. , 1973, Journal of Biological Chemistry.

[79]  Stuart K. Kim,et al.  A role for SIR-2.1 regulation of ER stress response genes in determining C. elegans life span. , 2005, Developmental cell.

[80]  D. Albert,et al.  Mitochondria, Calcium, and Calpain are Key Mediators of Resveratrol-Induced Apoptosis in Breast Cancer , 2007, Molecular Pharmacology.

[81]  Huan Wang,et al.  Epac-mediated activation of phospholipase C(epsilon) plays a critical role in beta-adrenergic receptor-dependent enhancement of Ca2+ mobilization in cardiac myocytes. , 2007, The Journal of biological chemistry.