Arrabal, SergioLucena, Miguel AngelCanduela, Miren JosuneRamos-Uriarte, AlmudenaRivera, PatriciaSerrano, AntoniaPavón, Francisco JavierDecara, JuanVargas, AntonioBaixeras, ElenaMartín-Rufián, MercedesMárquez, JavierFernández-Llébrez, PedroDe Roos, BaukjeGrandes, PedroRodríguez de Fonseca, FernandoSuárez, Juan2016-08-092016-08-092015-12-15Arrabal S, Lucena MA, Canduela MJ, Ramos-Uriarte A, Rivera P, Serrano A, et al. Pharmacological Blockade of Cannabinoid CB1 Receptors in Diet-Induced Obesity Regulates Mitochondrial Dihydrolipoamide Dehydrogenase in Muscle. PLoS ONE. 2015; 10(12):e0145244http://hdl.handle.net/10668/2333Journal Article; Research Support, Non-U.S. Gov't;Cannabinoid CB1 receptors peripherally modulate energy metabolism. Here, we investigated the role of CB1 receptors in the expression of glucose/pyruvate/tricarboxylic acid (TCA) metabolism in rat abdominal muscle. Dihydrolipoamide dehydrogenase (DLD), a flavoprotein component (E3) of α-ketoacid dehydrogenase complexes with diaphorase activity in mitochondria, was specifically analyzed. After assessing the effectiveness of the CB1 receptor antagonist AM251 (3 mg kg(-1), 14 days) on food intake and body weight, we could identified seven key enzymes from either glycolytic pathway or TCA cycle--regulated by both diet and CB1 receptor activity--through comprehensive proteomic approaches involving two-dimensional electrophoresis and MALDI-TOF/LC-ESI trap mass spectrometry. These enzymes were glucose 6-phosphate isomerase (GPI), triosephosphate isomerase (TPI), enolase (Eno3), lactate dehydrogenase (LDHa), glyoxalase-1 (Glo1) and the mitochondrial DLD, whose expressions were modified by AM251 in hypercaloric diet-induced obesity. Specifically, AM251 blocked high-carbohydrate diet (HCD)-induced expression of GPI, TPI, Eno3 and LDHa, suggesting a down-regulation of glucose/pyruvate/lactate pathways under glucose availability. AM251 reversed the HCD-inhibited expression of Glo1 and DLD in the muscle, and the DLD and CB1 receptor expression in the mitochondrial fraction. Interestingly, we identified the presence of CB1 receptors at the membrane of striate muscle mitochondria. DLD over-expression was confirmed in muscle of CB1-/- mice. AM251 increased the pyruvate dehydrogenase and glutathione reductase activity in C2C12 myotubes, and the diaphorase/oxidative activity in the mitochondria fraction. These results indicated an up-regulation of methylglyoxal and TCA cycle activity. Findings suggest that CB1 receptors in muscle modulate glucose/pyruvate/lactate pathways and mitochondrial oxidative activity by targeting DLD.enMúsculos abdominalesPeso corporalCannabinoidesMetabolismo energéticoGlucosaRatasObesidadMedical Subject Headings::Chemicals and Drugs::Enzymes and Coenzymes::Enzymes::Oxidoreductases::Oxidoreductases Acting on Aldehyde or Oxo Group Donors::Ketone Oxidoreductases::3-Methyl-2-Oxobutanoate Dehydrogenase (Lipoamide)Medical Subject Headings::Anatomy::Musculoskeletal System::Muscles::Muscle, Skeletal::Abdominal MusclesMedical Subject Headings::Organisms::Eukaryota::AnimalsMedical Subject Headings::Analytical, Diagnostic and Therapeutic Techniques and Equipment::Investigative Techniques::Anthropometry::Body Weights and Measures::Body Size::Body WeightMedical Subject Headings::Chemicals and Drugs::Organic Chemicals::Hydrocarbons::Terpenes::CannabinoidsMedical Subject Headings::Phenomena and Processes::Physiological Phenomena::Nutritional Physiological Phenomena::DietMedical Subject Headings::Chemicals and Drugs::Amino Acids, Peptides, and Proteins::Proteins::Flavoproteins::Dihydrolipoamide DehydrogenaseMedical Subject Headings::Phenomena and Processes::Chemical Phenomena::Biochemical Phenomena::Biochemical Processes::Down-RegulationMedical Subject Headings::Analytical, Diagnostic and Therapeutic Techniques and Equipment::Investigative Techniques::Electrochemical Techniques::ElectrophoresisMedical Subject Headings::Phenomena and Processes::Metabolic Phenomena::Metabolism::Energy MetabolismMedical Subject Headings::Chemicals and Drugs::Amino Acids, Peptides, and Proteins::Proteins::FlavoproteinsMedical Subject Headings::Chemicals and Drugs::Carbohydrates::Monosaccharides::Hexoses::GlucoseMedical Subject Headings::Chemicals and Drugs::Enzymes and Coenzymes::Enzymes::Oxidoreductases::Oxidoreductases Acting on Sulfur Group Donors::Glutathione ReductaseMedical Subject Headings::Chemicals and Drugs::Enzymes and Coenzymes::Enzymes::Oxidoreductases::Alcohol Oxidoreductases::Lactate Dehydrogenases::L-Lactate DehydrogenaseMedical Subject Headings::Chemicals and Drugs::Organic Chemicals::Carboxylic Acids::Hydroxy Acids::LactatesMedical Subject Headings::Organisms::Eukaryota::Animals::Chordata::Vertebrates::Mammals::Rodentia::Muridae::Murinae::MiceMedical Subject Headings::Anatomy::Cells::Cellular Structures::Intracellular Space::Cytoplasm::Cytoplasmic Structures::Organelles::MitochondriaMedical Subject Headings::Anatomy::Cells::Cellular Structures::Intracellular Space::Cytoplasm::Cytoplasmic Structures::Organelles::Mitochondria::Mitochondria, MuscleMedical Subject Headings::Anatomy::Tissues::Muscles::Muscle, Striated::Muscle, Skeletal::Muscle Fibers, SkeletalMedical Subject Headings::Analytical, Diagnostic and Therapeutic Techniques and Equipment::Diagnosis::Diagnostic Techniques and Procedures::Physical Examination::Body Constitution::Body Weights and Measures::Body Size::Body Weight::Overweight::ObesityMedical Subject Headings::Chemicals and Drugs::Inorganic Chemicals::Phosphorus Compounds::Phosphorus Acids::Phosphoric Acids::PhosphatesMedical Subject Headings::Chemicals and Drugs::Enzymes and Coenzymes::Enzymes::Lyases::Carbon-Oxygen Lyases::Hydro-Lyases::Phosphopyruvate HydrataseMedical Subject Headings::Chemicals and Drugs::Heterocyclic Compounds::Heterocyclic Compounds, 1-Ring::PiperidinesMedical Subject Headings::Disciplines and Occupations::Natural Science Disciplines::Chemistry::Biochemistry::ProteomicsMedical Subject Headings::Chemicals and Drugs::Heterocyclic Compounds::Heterocyclic Compounds, 1-Ring::Azoles::PyrazolesMedical Subject Headings::Chemicals and Drugs::Organic Chemicals::Aldehydes::Glyoxal::PyruvaldehydeMedical Subject Headings::Chemicals and Drugs::Amino Acids, Peptides, and Proteins::Proteins::Membrane Proteins::Receptors, Cell Surface::Receptors, G-Protein-Coupled::Receptors, Cannabinoid::Receptor, Cannabinoid, CB1Medical Subject Headings::Analytical, Diagnostic and Therapeutic Techniques and Equipment::Investigative Techniques::Chemistry Techniques, Analytical::Mass Spectrometry::Spectrometry, Mass, Matrix-Assisted Laser Desorption-IonizationMedical Subject Headings::Chemicals and Drugs::Organic Chemicals::Carboxylic Acids::Acids, Acyclic::Tricarboxylic AcidsMedical Subject Headings::Chemicals and Drugs::Enzymes and Coenzymes::Enzymes::Isomerases::Intramolecular Oxidoreductases::Aldose-Ketose Isomerases::Triose-Phosphate IsomeraseMedical Subject Headings::Phenomena and Processes::Chemical Phenomena::Chemical Processes::Biochemical Processes::Up-Regulationresearch article26671069open access10.1371/journal.pone.01452441932-6203PMC4682857