Casanova 2014 Biochim Biophys Acta

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Casanova E, Baselga-Escudero L, Ribas-Latre A, Arola-Arnal A, Bladé C, Arola L, Salvadó MJ (2014) Epigallocatechin gallate counteracts oxidative stress in docosahexaenoxic acid-treated myocytes. Biochim Biophys Acta 1837:783-91.

» PMID:24486445

Casanova E, Baselga-Escudero L, Ribas-Latre A, Arola-Arnal A, Blade C, Arola L, Salvado MJ (2014) Biochim Biophys Acta

Abstract: Skeletal muscle is a key organ of mammalian energy metabolism, and its mitochondria are multifunction organelles that are targets of dietary bioactive compounds. The goal of this work was to examine the regulation of mitochondrial dynamics, functionality and cell energy parameters using docosahexaenoic acid (DHA), epigallocatechin gallate (EGCG) and a combination of both in L6 myocytes. Compounds (at 25μM) were incubated for 4 hours. Cells cultured with DHA displayed a less oxygen consumption with a higher ADP/ATP ratio levels concomitant with downregulation of Cox and Ant1 genes expression. The disruption of energetic homeostasis by DHA, increase intracellular reactive oxygen species (ROS) levels and decrease mitochondrial membrane potential. The defence mechanism to counteract the excess of ROS production was by the upregulation of Ucp2, Ucp3 and Mnsod genes expression. Moreover myocytes cultured with DHA had a higher mitochondrial mass with a higher proportion of large and elongated mitochondria, whereas the fission genes Drp1 and Fiss1 and the fusion gene Mfn2 were downregulated. In myocytes co-incubated with DHA and EGCG; ROS levels and the ADP/ATP ratio were similar to untreated myocytes and the decrease of oxygen consumption, higher mitochondrial mass and the overexpression of Ucp2 and Ucp3 genes were similar to the DHA-treated cells with also higher amount of mitochondrial DNA, and reduced Drp1, Fiss1 gene expression levels. In conclusion the addition of EGCG to DHA returned the cells to the control conditions in terms of mitochondrial morphology, energy and redox status, which were unbalanced in the DHA-treated myocytes.

Keywords: 2’, 7’-dichlorofluorescin diacetate, AFU, Ant 1, Antioxidants, Cox, DCFH-DA, DHA, DMEM, Docosahexaenoic acid, Drp1, Dulbecco’s Modified Eagle’s Medium, ETC, ET-pathway, Electorn transport capapcity, Electron transport chain, Epigallocatechin gallate, FBS, FCCP, Fiss1, Gapdh, MMP, Mfn2, Mitochondria, Mnsod, Opa1, PUFAs, ROS, ROX, Reactive oxygen species, Rhd123, Rhodamine 123, Ucp2, Ucp3, adenine nucleotide translocase 1, arbitrary fluorescence units, carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone, cytochrome C oxidase subunit V;Cs, citrate synthase, docosahexaenoic acid, dynamin-related protein 1;EGCG, Epigallocatechin-3-gallate, fetal bovine serum, glyceraldehydes-3-phosphate dehydrogenase;LDH, Lactate dehydrogenase, manganese superoxide dismutase, mitochondrial DNA, mitochondrial fission 1 protein;Fluo-3AM, Fluo-3 acetoxymethyl ester, mitochondrial membrane potential, mitofusin 2, mtDNA, nDNA, nuclear DNA;Nd3, NADH-dehydrogenase subunit 3;OXPHOS, oxidative phosphorylation, optic atrophy 1, polyunsaturated fatty acids, reactive oxygen species, residual oxygen consumption, skeletal muscle, uncoupling protein 2, uncoupling protein 3

O2k-Network Lab: ES Tarragona Arola L


Labels: MiParea: Respiration, Pharmacology;toxicology 

Stress:Cell death  Organism: Rat  Tissue;cell: Skeletal muscle, Other cell lines  Preparation: Intact cells 

Regulation: mt-Membrane potential  Coupling state: LEAK, ROUTINE, ET  Pathway:HRR: Oxygraph-2k