Epigallocatechin gallate counteracts oxidative stress in docosahexaenoxic acid-treated myocytes.
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 4h. Cells cultured with DHA displayed less oxygen consumption with higher ADP/ATP ratio levels concomitant with downregulation of Cox and Ant1 gene expression. The disruption of energetic homeostasis by DHA, increases intracellular reactive oxygen species (ROS) levels and decreases mitochondrial membrane potential. The defence mechanism to counteract the excess of ROS production was by the upregulation of Ucp2, Ucp3 and MnSod gene 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 adenosine diphosphate (ADP)/adenosine triphosphate (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 a higher amount of mitochondrial deoxyribonucleic acid (DNA), and reduced Drp1 and 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.
Grup de Nutrigenòmica, Departament de Bioquimica i Biotecnologia, Universitat Rovira i Virgili, Campus Sescel·lades, 43007 Tarragona, Spain.
Ester
E
Casanova
Grup de Nutrigenòmica, Departament de Bioquimica i Biotecnologia, Universitat Rovira i Virgili, Campus Sescel·lades, 43007 Tarragona, Spain.
Laura
L
Baselga-Escudero
Grup de Nutrigenòmica, Departament de Bioquimica i Biotecnologia, Universitat Rovira i Virgili, Campus Sescel·lades, 43007 Tarragona, Spain.
Aleix
A
Ribas-Latre
Grup de Nutrigenòmica, Departament de Bioquimica i Biotecnologia, Universitat Rovira i Virgili, Campus Sescel·lades, 43007 Tarragona, Spain.
Anna
A
Arola-Arnal
Grup de Nutrigenòmica, Departament de Bioquimica i Biotecnologia, Universitat Rovira i Virgili, Campus Sescel·lades, 43007 Tarragona, Spain.
Cinta
C
Bladé
Grup de Nutrigenòmica, Departament de Bioquimica i Biotecnologia, Universitat Rovira i Virgili, Campus Sescel·lades, 43007 Tarragona, Spain.
Lluís
L
Arola
Grup de Nutrigenòmica, Departament de Bioquimica i Biotecnologia, Universitat Rovira i Virgili, Campus Sescel·lades, 43007 Tarragona, Spain. Electronic address: mariajosepa.salvado@urv.cat.
M Josepa
MJ
Salvadó
D000818:Animals; D002118:Calcium / Q000378:metabolism; D002392:Catechin / Q000031:analogs & derivatives* / Q000494:pharmacology; D002478:Cells, Cultured; D004281:Docosahexaenoic Acids / Q000494:pharmacology*; D008931:Mitochondria, Muscle / Q000187:drug effects / Q000378:metabolism; D018482:Muscle, Skeletal / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D010085:Oxidative Phosphorylation; D018384:Oxidative Stress / Q000187:drug effects*; D017382:Reactive Oxygen Species / Q000378:metabolism
D017382:Reactive Oxygen Species; D004281:Docosahexaenoic Acids; D002392:Catechin; C045651:epigallocatechin gallate; D002118:Calcium
Antioxidant; Docosahexaenoic acid; Epigallocatechin gallate; Mitochondria; Reactive oxygen species; Skeletal muscle