title: Novel targets of sulforaphane in primary cardiomyocytes identified by proteomic analysis.
abstract: Cardiovascular diseases represent the main cause of mortality in the industrialized world and the identification of effective preventive strategies is of fundamental importance. Sulforaphane, an isothiocyanate from cruciferous vegetables, has been shown to up-regulate phase II enzymes in cardiomyocytes and counteract oxidative stress-induced apoptosis. Aim of the present study was the identification and characterization of novel sulforaphane targets in cardiomyocytes applying a proteomic approach. Two-dimensional gel electrophoresis and mass spectrometry were used to generate protein profiles of primary neonatal rat cardiomyocytes treated and untreated with 5 µM sulforaphane for 1-48 h. According to image analysis, 64 protein spots were found as differentially expressed and their functional correlations were investigated using the MetaCore program. We mainly focused on 3 proteins: macrophage migration inhibitory factor (MIF), CLP36 or Elfin, and glyoxalase 1, due to their possible involvement in cardioprotection. Validation of the time-dependent differential expression of these proteins was performed by western blotting. In particular, to gain insight into the cardioprotective role of the modulation of glyoxalase 1 by sulforaphane, further experiments were performed using methylglyoxal to mimic glycative stress. Sulforaphane was able to counteract methylglyoxal-induced apoptosis, ROS production, and glycative stress, likely through glyoxalase 1 up-regulation. In this study, we reported for the first time new molecular targets of sulforaphane, such as MIF, CLP36 and glyoxalase 1. In particular, we gave new insights into the anti-glycative role of sulforaphane in cardiomyocytes, confirming its pleiotropic behavior in counteracting cardiovascular diseases.
authors.affiliation: Department for Life Quality Studies, Alma Mater Studiorum - University of Bologna, Bologna, Italy.
authors.forename: Cristina
authors.initials: C
authors.lastname: Angeloni
authors.affiliation: Department of Pharmacy and Biotechnology, Alma Mater Studiorum - University of Bologna, Bologna, Italy.
authors.forename: Silvia
authors.initials: S
authors.lastname: Turroni
authors.affiliation: Functional Proteomics Laboratory, Department of Life Sciences, University of Siena, Siena, Italy.
authors.forename: Laura
authors.initials: L
authors.lastname: Bianchi
authors.affiliation: Department for Life Quality Studies, Alma Mater Studiorum - University of Bologna, Bologna, Italy.
authors.forename: Daniele
authors.initials: D
authors.lastname: Fabbri
authors.affiliation: Department for Life Quality Studies, Alma Mater Studiorum - University of Bologna, Bologna, Italy.
authors.forename: Elisa
authors.initials: E
authors.lastname: Motori
authors.affiliation: Department for Life Quality Studies, Alma Mater Studiorum - University of Bologna, Bologna, Italy.
authors.forename: Marco
authors.initials: M
authors.lastname: Malaguti
authors.affiliation: Department for Life Quality Studies, Alma Mater Studiorum - University of Bologna, Bologna, Italy.
authors.forename: Emanuela
authors.initials: E
authors.lastname: Leoncini
authors.affiliation: Department of Surgical, Medical, Dental and Morphological Sciences with interest in Transplant, Oncology and Regenerative Medicine, University of Modena and Reggio Emilia, Modena, Italy.
authors.forename: Tullia
authors.initials: T
authors.lastname: Maraldi
authors.affiliation: Functional Proteomics Laboratory, Department of Life Sciences, University of Siena, Siena, Italy.
authors.forename: Luca
authors.initials: L
authors.lastname: Bini
authors.affiliation: Department of Pharmacy and Biotechnology, Alma Mater Studiorum - University of Bologna, Bologna, Italy.
authors.forename: Patrizia
authors.initials: P
authors.lastname: Brigidi
authors.affiliation: Department for Life Quality Studies, Alma Mater Studiorum - University of Bologna, Bologna, Italy.
authors.forename: Silvana
authors.initials: S
authors.lastname: Hrelia
mesh_terms: D000818:Animals; D016588:Anticarcinogenic Agents / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects; D002316:Cardiotonic Agents / Q000494:pharmacology*; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D017879:Isothiocyanates / Q000494:pharmacology*; D009124:Muscle Proteins / Q000096:biosynthesis*; D032383:Myocytes, Cardiac / Q000166:cytology / Q000378:metabolism*; D040901:Proteomics; D011765:Pyruvaldehyde / Q000633:toxicity; D051381:Rats; D017208:Rats, Wistar; D017382:Reactive Oxygen Species / Q000378:metabolism; D013454:Sulfoxides
chemical_list: D016588:Anticarcinogenic Agents; D002316:Cardiotonic Agents; D017879:Isothiocyanates; D009124:Muscle Proteins; D017382:Reactive Oxygen Species; D013454:Sulfoxides; D011765:Pyruvaldehyde; C016766:sulforaphane