Novel Targets of Sulforaphane in Primary Cardiomyocytes Identified by Proteomic Analysis
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{"title"=>"Novel targets of sulforaphane in primary cardiomyocytes identified by proteomic analysis", "type"=>"journal", "authors"=>[{"first_name"=>"Cristina", "last_name"=>"Angeloni", "scopus_author_id"=>"6602199774"}, {"first_name"=>"Silvia", "last_name"=>"Turroni", "scopus_author_id"=>"16319673700"}, {"first_name"=>"Laura", "last_name"=>"Bianchi", "scopus_author_id"=>"57201067549"}, {"first_name"=>"Daniele", "last_name"=>"Fabbri", "scopus_author_id"=>"57201193823"}, {"first_name"=>"Elisa", "last_name"=>"Motori", "scopus_author_id"=>"39262401200"}, {"first_name"=>"Marco", "last_name"=>"Malaguti", "scopus_author_id"=>"23009163900"}, {"first_name"=>"Emanuela", "last_name"=>"Leoncini", "scopus_author_id"=>"6507905782"}, {"first_name"=>"Tullia", "last_name"=>"Maraldi", "scopus_author_id"=>"6507881128"}, {"first_name"=>"Luca", "last_name"=>"Bini", "scopus_author_id"=>"7005544275"}, {"first_name"=>"Patrizia", "last_name"=>"Brigidi", "scopus_author_id"=>"7003712308"}, {"first_name"=>"Silvana", "last_name"=>"Hrelia", "scopus_author_id"=>"7006925536"}], "year"=>2013, "source"=>"PLoS ONE", "identifiers"=>{"sgr"=>"84892609715", "doi"=>"10.1371/journal.pone.0083283", "pui"=>"372133172", "issn"=>"19326203", "pmid"=>"24349480", "isbn"=>"1932-6203 (Electronic)\\r1932-6203 (Linking)", "scopus"=>"2-s2.0-84892609715"}, "id"=>"26c79f0f-e43f-3544-aaf9-1355b024d07b", "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.", "link"=>"http://www.mendeley.com/research/novel-targets-sulforaphane-primary-cardiomyocytes-identified-proteomic-analysis", "reader_count"=>26, "reader_count_by_academic_status"=>{"Unspecified"=>2, "Researcher"=>6, "Student > Ph. D. Student"=>6, "Student > Postgraduate"=>2, "Student > Master"=>4, "Other"=>2, "Student > Bachelor"=>2, "Lecturer"=>1, "Professor"=>1}, "reader_count_by_user_role"=>{"Unspecified"=>2, "Researcher"=>6, "Student > Ph. D. Student"=>6, "Student > Postgraduate"=>2, "Student > Master"=>4, "Other"=>2, "Student > Bachelor"=>2, "Lecturer"=>1, "Professor"=>1}, "reader_count_by_subject_area"=>{"Unspecified"=>2, "Biochemistry, Genetics and Molecular Biology"=>5, "Agricultural and Biological Sciences"=>7, "Medicine and Dentistry"=>10, "Chemistry"=>2}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>10}, "Chemistry"=>{"Chemistry"=>2}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>7}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>5}, "Unspecified"=>{"Unspecified"=>2}}, "reader_count_by_country"=>{"United States"=>1}, "group_count"=>0}

Scopus | Further Information

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/1311088"], "description"=>"<p>The top score transcription regulation network initiated through activation of SP1 is shown. Network proteins are visualized by proper symbols, which specify the functional nature of the protein (network caption). Green and gray arrows indicate positive and unspecified effects, respectively. Red and blue circles indicate up- and down-regulated proteins following SF treatment, respectively. ‘Checkerboard’ color indicates mixed expression between files.</p>", "links"=>[], "tags"=>["differentially"], "article_id"=>874633, "categories"=>["Biological Sciences"], "users"=>["Cristina Angeloni", "Silvia Turroni", "Laura Bianchi", "Daniele Fabbri", "Elisa Motori", "Marco Malaguti", "Emanuela Leoncini", "Tullia Maraldi", "Luca Bini", "Patrizia Brigidi", "Silvana Hrelia"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0083283.g003", "stats"=>{"downloads"=>1, "page_views"=>11, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Network_analysis_of_differentially_expressed_proteins_/874633", "title"=>"Network analysis of differentially expressed proteins.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-12-11 03:17:41"}
  • {"files"=>["https://ndownloader.figshare.com/files/1311085"], "description"=>"<p>(<b>A</b>) PCA plot of the protein spot maps of cardiomyocytes untreated (DMSO vehicle control; circles) and after exposure to SF for 1 (triangles), 6 (squares), 24 (diamonds) and 48 h (crosses). Log-transformed data were used. Each symbol represents a 2-DE gel from each treatment series. First and second ordination axes are plotted explaining 36.8 and 17.8% of the overall variance in the dataset, respectively. (<b>B</b>) Two-way hierarchical clustering of the 64 differentially expressed protein spots between SF-treated cardiomyocytes and DMSO vehicle control. Pearson's dissimilarity as distance measure and Ward's method for linkage analysis were used. Log<sub>2</sub> ratios are colour coded as indicated. Names of the identified protein spots are shown on the right (see <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0083283#pone.0083283.s006\" target=\"_blank\">Table S1</a>). A larger version of this image with gene names and spot IDs in brackets is available as <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0083283#pone.0083283.s001\" target=\"_blank\">Figure S1</a>. (<b>C</b>-<b>E</b>) Functional categories by Gene Ontology (GO) analysis of the identified proteins. Classification was performed according to keyword categories [(<b>C</b>) biological process, (<b>D</b>) cellular component, (<b>E</b>) molecular function]. When proteins were associated with more than one functional category, one GO term was chosen arbitrarily. NA, not available. ER, endoplasmic reticulum.</p>", "links"=>[], "tags"=>["proteomic"], "article_id"=>874630, "categories"=>["Biological Sciences"], "users"=>["Cristina Angeloni", "Silvia Turroni", "Laura Bianchi", "Daniele Fabbri", "Elisa Motori", "Marco Malaguti", "Emanuela Leoncini", "Tullia Maraldi", "Luca Bini", "Patrizia Brigidi", "Silvana Hrelia"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0083283.g002", "stats"=>{"downloads"=>0, "page_views"=>6, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Multivariate_analysis_of_proteomic_data_/874630", "title"=>"Multivariate analysis of proteomic data.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-12-11 03:17:41"}
  • {"files"=>["https://ndownloader.figshare.com/files/1311099", "https://ndownloader.figshare.com/files/1311100", "https://ndownloader.figshare.com/files/1311101", "https://ndownloader.figshare.com/files/1311102", "https://ndownloader.figshare.com/files/1311103", "https://ndownloader.figshare.com/files/1311104", "https://ndownloader.figshare.com/files/1311105", "https://ndownloader.figshare.com/files/1311106", "https://ndownloader.figshare.com/files/1311107"], "description"=>"<div><p>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.</p> </div>", "links"=>[], "tags"=>["targets", "sulforaphane", "cardiomyocytes", "proteomic"], "article_id"=>874644, "categories"=>["Biological Sciences"], "users"=>["Cristina Angeloni", "Silvia Turroni", "Laura Bianchi", "Daniele Fabbri", "Elisa Motori", "Marco Malaguti", "Emanuela Leoncini", "Tullia Maraldi", "Luca Bini", "Patrizia Brigidi", "Silvana Hrelia"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0083283.s001", "https://dx.doi.org/10.1371/journal.pone.0083283.s002", "https://dx.doi.org/10.1371/journal.pone.0083283.s003", "https://dx.doi.org/10.1371/journal.pone.0083283.s004", "https://dx.doi.org/10.1371/journal.pone.0083283.s005", "https://dx.doi.org/10.1371/journal.pone.0083283.s006", "https://dx.doi.org/10.1371/journal.pone.0083283.s007", "https://dx.doi.org/10.1371/journal.pone.0083283.s008", "https://dx.doi.org/10.1371/journal.pone.0083283.s009"], "stats"=>{"downloads"=>35, "page_views"=>10, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Novel_Targets_of_Sulforaphane_in_Primary_Cardiomyocytes_Identified_by_Proteomic_Analysis_/874644", "title"=>"Novel Targets of Sulforaphane in Primary Cardiomyocytes Identified by Proteomic Analysis", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2013-12-11 03:17:41"}
  • {"files"=>["https://ndownloader.figshare.com/files/1311096"], "description"=>"<p>Cardiomyocytes were treated with 5 µM SF for 24 h before addition of 1 mM MG. Cell lysates were submitted to the ABTS radical cation decolorization assay and the antioxidant activity of the cells were expressed as mean ± SD of µmol of trolox antioxidant activity per mg of protein. p<0.05 vs Control; ° p<0.05 vs MG.</p>", "links"=>[], "tags"=>["cardiomyocytes", "antioxidant"], "article_id"=>874641, "categories"=>["Biological Sciences"], "users"=>["Cristina Angeloni", "Silvia Turroni", "Laura Bianchi", "Daniele Fabbri", "Elisa Motori", "Marco Malaguti", "Emanuela Leoncini", "Tullia Maraldi", "Luca Bini", "Patrizia Brigidi", "Silvana Hrelia"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0083283.g008", "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_SF_effects_on_cardiomyocytes_total_antioxidant_activity_/874641", "title"=>"SF effects on cardiomyocytes total antioxidant activity.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-12-11 03:17:41"}
  • {"files"=>["https://ndownloader.figshare.com/files/1311094"], "description"=>"<p>Cardiomyocytes were treated with 5 µM SF for 24 h before addition of 1 mM MG. (<b>A</b>) Intracellular ROS levels were determined with the peroxide-sensitive probe DCFH-DA. (<b>B</b>) Cytosolic ROS levels were determined with DHE. (<b>C</b>) Mitochondrial ROS levels were determined with MitoSox. Data are expressed as a percentage compared to control and are presented as mean ± SD, n = 4 in each group,* p<0.05 vs Control; ° p<0.05 vs MG.</p>", "links"=>[], "tags"=>["assays", "sf", "mg-induced", "ros"], "article_id"=>874639, "categories"=>["Biological Sciences"], "users"=>["Cristina Angeloni", "Silvia Turroni", "Laura Bianchi", "Daniele Fabbri", "Elisa Motori", "Marco Malaguti", "Emanuela Leoncini", "Tullia Maraldi", "Luca Bini", "Patrizia Brigidi", "Silvana Hrelia"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0083283.g007", "stats"=>{"downloads"=>1, "page_views"=>9, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Fluorimetric_assays_of_SF_effect_on_MG_induced_ROS_generation_/874639", "title"=>"Fluorimetric assays of SF effect on MG-induced ROS generation.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-12-11 03:17:41"}
  • {"files"=>["https://ndownloader.figshare.com/files/1311093"], "description"=>"<p>Cardiomyocytes were treated with 5 µM SF for 24 h before the addition of 1 mM MG. Cytosolic ROS production was measured using DHE and mitochondrial ROS production by MitoSOX staining. Scale bar: 10 µm.</p>", "links"=>[], "tags"=>["microscopy", "sf", "mg-induced", "ros"], "article_id"=>874638, "categories"=>["Biological Sciences"], "users"=>["Cristina Angeloni", "Silvia Turroni", "Laura Bianchi", "Daniele Fabbri", "Elisa Motori", "Marco Malaguti", "Emanuela Leoncini", "Tullia Maraldi", "Luca Bini", "Patrizia Brigidi", "Silvana Hrelia"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0083283.g006", "stats"=>{"downloads"=>0, "page_views"=>2, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Confocal_microscopy_analysis_of_SF_effect_on_MG_induced_ROS_production_/874638", "title"=>"Confocal microscopy analysis of SF effect on MG-induced ROS production.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-12-11 03:17:41"}
  • {"files"=>["https://ndownloader.figshare.com/files/1311090"], "description"=>"<p>Cardiomyocytes were treated with 5 µM SF for 24 h before addition of 1 mM MG. (<b>A</b>) Cell viability was assessed by MTT assay and reported as percent cell viability in comparison to control. (<b>B</b>) Caspase-3 activity was measured spectrofluorimetrically by hydrolysis of the peptide substrate acetyl-Asp-Glu-Val-Asp-7-amido-4-methylcoumarin (Ac-DEVD-AMC). (<b>C</b>) Cell lysates were immunoblotted with caspase-3 antibody that detects both full length caspase-3 (35 kDa) and the large fragment of caspase-3 resulting from cleavage (17 kDa). Data are presented as mean ± SD, n = 4 in each group,* p<0.05 vs Control; ° p<0.05 vs MG.</p>", "links"=>[], "tags"=>["mg-induced"], "article_id"=>874635, "categories"=>["Biological Sciences"], "users"=>["Cristina Angeloni", "Silvia Turroni", "Laura Bianchi", "Daniele Fabbri", "Elisa Motori", "Marco Malaguti", "Emanuela Leoncini", "Tullia Maraldi", "Luca Bini", "Patrizia Brigidi", "Silvana Hrelia"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0083283.g005", "stats"=>{"downloads"=>0, "page_views"=>6, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_SF_protection_against_MG_induced_damage_/874635", "title"=>"SF protection against MG-induced damage.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-12-11 03:17:41"}
  • {"files"=>["https://ndownloader.figshare.com/files/1311089"], "description"=>"<p>Cardiomyocytes were treated with 5 μM SF for 1-48 h followed by western blot analysis for MIF (<b>A</b>), CLP36 (<b>B</b>), and GLO1 (<b>C</b>). Representative bands and raw densitometric values of the corresponding protein spots from proteomic analysis are shown. Each bar represents the spot quantity as determined by PDQuest (mean ± SD). At least three biological replicates with two technical replicates were run for each tested condition.</p>", "links"=>[], "tags"=>["ms"], "article_id"=>874634, "categories"=>["Biological Sciences"], "users"=>["Cristina Angeloni", "Silvia Turroni", "Laura Bianchi", "Daniele Fabbri", "Elisa Motori", "Marco Malaguti", "Emanuela Leoncini", "Tullia Maraldi", "Luca Bini", "Patrizia Brigidi", "Silvana Hrelia"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0083283.g004", "stats"=>{"downloads"=>0, "page_views"=>2, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Validation_of_MS_results_by_immunoblotting_/874634", "title"=>"Validation of MS results by immunoblotting.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-12-11 03:17:41"}
  • {"files"=>["https://ndownloader.figshare.com/files/1311084"], "description"=>"<p>IEF performed on IPG strips (13 cm, 3-10 linear pH gradient) was followed by second dimension separation on a 15% polyacrylamide gel. 2-DE gel was silver-stained. Identified spots, showing a statistically significant difference of at least 1.5-fold between SF-treated cardiomyocytes at different time points [1 (B), 6 (C), 24 (D) and 48 (E) h] and vehicle control (A), are numbered (<a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0083283#pone.0083283.s006\" target=\"_blank\">Table S1</a>). Circles and squares indicate up- and down-regulated protein spots, respectively. (A) Diamonds indicate protein spots with opposite trends of expression among the time points of exposure to SF.</p>", "links"=>[], "tags"=>["cardiomyocytes", "treated"], "article_id"=>874629, "categories"=>["Biological Sciences"], "users"=>["Cristina Angeloni", "Silvia Turroni", "Laura Bianchi", "Daniele Fabbri", "Elisa Motori", "Marco Malaguti", "Emanuela Leoncini", "Tullia Maraldi", "Luca Bini", "Patrizia Brigidi", "Silvana Hrelia"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0083283.g001", "stats"=>{"downloads"=>0, "page_views"=>1, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_2_DE_protein_profile_of_cardiomyocytes_treated_with_SF_/874629", "title"=>"2-DE protein profile of cardiomyocytes treated with SF.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-12-11 03:17:41"}
  • {"files"=>["https://ndownloader.figshare.com/files/1311097"], "description"=>"<p>(<b>A</b>) Cardiomyocytes were treated with 5 µM SF for different time intervals (1-48 h) and GLO1 enzyme activity was measured by a spectrophotometric method. (<b>B</b>) Cardiomyocytes were treated with 5 µM SF for 24 h before addition of 1 mM MG, and GLO1 enzyme activity was measured by a spectrophotometric method. (<b>C</b>) Cardiomyocytes were treated with 5 µM SF for 24 h before addition of 1 mM MG, and AGE formation was evaluated by AGE-ELISA using a specific anti-AGE antibody. Data are presented as mean ± SD, n = 4 in each group, * p<0.05 vs Control; ° p<0.05 vs MG.</p>", "links"=>[], "tags"=>["sf", "mg-induced"], "article_id"=>874642, "categories"=>["Biological Sciences"], "users"=>["Cristina Angeloni", "Silvia Turroni", "Laura Bianchi", "Daniele Fabbri", "Elisa Motori", "Marco Malaguti", "Emanuela Leoncini", "Tullia Maraldi", "Luca Bini", "Patrizia Brigidi", "Silvana Hrelia"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0083283.g009", "stats"=>{"downloads"=>1, "page_views"=>7, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Effect_of_SF_against_MG_induced_glycation_/874642", "title"=>"Effect of SF against MG-induced glycation.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-12-11 03:17:41"}

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Relative Metric

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