Sulforaphane Restores Cellular Glutathione Levels and Reduces Chronic Periodontitis Neutrophil Hyperactivity In Vitro
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{"title"=>"Sulforaphane Restores Cellular Glutathione Levels and Reduces Chronic Periodontitis Neutrophil Hyperactivity In Vitro", "type"=>"journal", "authors"=>[{"first_name"=>"Irundika H.K.", "last_name"=>"Dias", "scopus_author_id"=>"35083345400"}, {"first_name"=>"Ian L.C.", "last_name"=>"Chapple", "scopus_author_id"=>"7003386333"}, {"first_name"=>"Mike", "last_name"=>"Milward", "scopus_author_id"=>"9535750500"}, {"first_name"=>"Melissa M.", "last_name"=>"Grant", "scopus_author_id"=>"7401439900"}, {"first_name"=>"Eric", "last_name"=>"Hill", "scopus_author_id"=>"22950882700"}, {"first_name"=>"James", "last_name"=>"Brown", "scopus_author_id"=>"24578793300"}, {"first_name"=>"Helen R.", "last_name"=>"Griffiths", "scopus_author_id"=>"57193264946"}], "year"=>2013, "source"=>"PLoS ONE", "identifiers"=>{"pui"=>"369200041", "issn"=>"19326203", "isbn"=>"1932-6203 (Electronic)\\r1932-6203 (Linking)", "doi"=>"10.1371/journal.pone.0066407", "scopus"=>"2-s2.0-84879479555", "pmid"=>"23826097", "sgr"=>"84879479555"}, "id"=>"7cc28993-b7ed-308f-a02c-33cdb66339ef", "abstract"=>"The production of high levels of reactive oxygen species by neutrophils is associated with the local and systemic destructive phenotype found in the chronic inflammatory disease periodontitis. In the present study, we investigated the ability of sulforaphane (SFN) to restore cellular glutathione levels and reduce the hyperactivity of circulating neutrophils associated with chronic periodontitis. Using differentiated HL60 cells as a neutrophil model, here we show that generation of extracellular O2 (. -) by the nicotinamide adenine dinucleotide (NADPH) oxidase complex is increased by intracellular glutathione depletion. This may be attributed to the upregulation of thiol regulated acid sphingomyelinase driven lipid raft formation. Intracellular glutathione was also lower in primary neutrophils from periodontitis patients and, consistent with our previous findings, patients neutrophils were hyper-reactive to stimuli. The activity of nuclear factor erythroid-2-related factor 2 (Nrf2), a master regulator of the antioxidant response, is impaired in circulating neutrophils from chronic periodontitis patients. Although patients' neutrophils exhibit a low reduced glutathione (GSH)/oxidised glutathione (GSSG) ratio and a higher total Nrf2 level, the DNA-binding activity of nuclear Nrf2 remained unchanged relative to healthy controls and had reduced expression of glutamate cysteine ligase catalytic (GCLC), and modifier (GCLM) subunit mRNAs, compared to periodontally healthy subjects neutrophils. Pre-treatment with SFN increased expression of GCLC and GCM, improved intracellular GSH/GSSG ratios and reduced agonist-activated extracellular O2 (. -) production in both dHL60 and primary neutrophils from patients with periodontitis and controls. These findings suggest that a deficiency in Nrf2-dependent pathways may underpin susceptibility to hyper-reactivity in circulating primary neutrophils during chronic periodontitis.", "link"=>"http://www.mendeley.com/research/sulforaphane-restores-cellular-glutathione-levels-reduces-chronic-periodontitis-neutrophil-hyperacti", "reader_count"=>32, "reader_count_by_academic_status"=>{"Unspecified"=>1, "Professor > Associate Professor"=>1, "Researcher"=>8, "Student > Doctoral Student"=>4, "Student > Ph. D. Student"=>6, "Student > Postgraduate"=>1, "Student > Master"=>3, "Other"=>1, "Student > Bachelor"=>6, "Professor"=>1}, "reader_count_by_user_role"=>{"Unspecified"=>1, "Professor > Associate Professor"=>1, "Researcher"=>8, "Student > Doctoral Student"=>4, "Student > Ph. D. Student"=>6, "Student > Postgraduate"=>1, "Student > Master"=>3, "Other"=>1, "Student > Bachelor"=>6, "Professor"=>1}, "reader_count_by_subject_area"=>{"Unspecified"=>2, "Biochemistry, Genetics and Molecular Biology"=>2, "Nursing and Health Professions"=>1, "Agricultural and Biological Sciences"=>10, "Medicine and Dentistry"=>12, "Neuroscience"=>1, "Pharmacology, Toxicology and Pharmaceutical Science"=>2, "Chemistry"=>2}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>12}, "Neuroscience"=>{"Neuroscience"=>1}, "Chemistry"=>{"Chemistry"=>2}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>10}, "Nursing and Health Professions"=>{"Nursing and Health Professions"=>1}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>2}, "Unspecified"=>{"Unspecified"=>2}, "Pharmacology, Toxicology and Pharmaceutical Science"=>{"Pharmacology, Toxicology and Pharmaceutical Science"=>2}}, "reader_count_by_country"=>{"United States"=>1, "Poland"=>1}, "group_count"=>1}

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/1098513"], "description"=>"<p>The pathway leading to the formation of GSH by the action of γ-glutamylcysteine synthetase (γGCS) is blocked by buthionine sulfoximine (BSO), inducing artificial stress condition in dHL60 cells. Decreased cellular GSH/GSSG ratio may activate redox sensitive enzymes such as ASMases and nuclear translocation of Nrf2. ASMases may favour lipid raft formation and thereby clustering active NADPH oxidase complexes in the outer membrane, subsequently liberating extracellular O<sub>2</sub><sup>.-</sup>.</p>", "links"=>[], "tags"=>["up-regulation", "nadph", "oxidase", "redox"], "article_id"=>729484, "categories"=>["Biological Sciences"], "users"=>["Irundika H. K. Dias", "Ian L. C. Chapple", "Mike Milward", "Melissa M. Grant", "Eric Hill", "James Brown", "Helen R. Griffiths"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0066407.g006", "stats"=>{"downloads"=>1, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_A_possible_mechanism_for_the_up_regulation_of_NADPH_oxidase_activity_under_redox_stress_/729484", "title"=>"A possible mechanism for the up-regulation of NADPH oxidase activity under redox stress.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-06-24 02:02:16"}
  • {"files"=>["https://ndownloader.figshare.com/files/1098510"], "description"=>"<p>Peak relative light unit (RLU) values for neutrophils stimulated with (A) 1µM fMLP, (B) opsonised <i>S. aureus</i> (300 bacteria/neutrophil) (C) <i>F. nucleatum</i> bacteria x100/neutrophil and (D) 10nM PMA in the presence or absence of SFN treatment were plotted and significant differences were calculated using Tukey’s multiple-comparison test, where, * p<0.05 and **p<0.01.</p>", "links"=>[], "tags"=>["pre-incubation", "decreases", "neutrophil", "respiratory"], "article_id"=>729481, "categories"=>["Biological Sciences"], "users"=>["Irundika H. K. Dias", "Ian L. C. Chapple", "Mike Milward", "Melissa M. Grant", "Eric Hill", "James Brown", "Helen R. Griffiths"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0066407.g004", "stats"=>{"downloads"=>1, "page_views"=>6, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_SFN_pre_incubation_decreases_neutrophil_respiratory_burst_/729481", "title"=>"SFN pre-incubation decreases neutrophil respiratory burst.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-06-24 02:02:16"}
  • {"files"=>["https://ndownloader.figshare.com/files/1098508"], "description"=>"<p>(A) BSO (10µM) pre-treated and untreated dHL60 cells were treated with or without desipramine (10 µM, 1 hour) and 5µM SFN. Cells (1×10<sup>7</sup>) were collected on ice-cold PBS, pelleted and lysed before analysing ASMase activity. (B) LRs from dHL60 cells were extracted with MNE buffer containing 1% triton X-100 and applied to discontinuous sucrose gradients, as described in Methods. Proteins were extracted and immunoblotted using anti-flotillin-1 as the raft protein detecting antibody. (C) Quantification of flotillin in lipid raft fractions 3 and 4 compared to total expression.</p>", "links"=>[], "tags"=>["oxidative", "asmase", "lr"], "article_id"=>729479, "categories"=>["Biological Sciences"], "users"=>["Irundika H. K. Dias", "Ian L. C. Chapple", "Mike Milward", "Melissa M. Grant", "Eric Hill", "James Brown", "Helen R. Griffiths"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0066407.g002", "stats"=>{"downloads"=>0, "page_views"=>1, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Effects_of_oxidative_stress_on_ASMase_activity_and_LR_rearrangement_/729479", "title"=>"Effects of oxidative stress on ASMase activity and LR rearrangement.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-06-24 02:02:16"}
  • {"files"=>["https://ndownloader.figshare.com/files/1098509"], "description"=>"<p>GSH (A) and GGSG (B) concentrations, along with the GSH/GSSG ratio (C) were measured using the DTNB recycling assay and expressed relative to protein content. Significant differences were calculated with Tukey’s multiple-comparison test where*P<0.05, and ***P<0.001.</p>", "links"=>[], "tags"=>["pre-incubation"], "article_id"=>729480, "categories"=>["Biological Sciences"], "users"=>["Irundika H. K. Dias", "Ian L. C. Chapple", "Mike Milward", "Melissa M. Grant", "Eric Hill", "James Brown", "Helen R. Griffiths"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0066407.g003", "stats"=>{"downloads"=>1, "page_views"=>6, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_SFN_pre_incubation_increases_the_GSH_GSSG_ratio_in_primary_neutrophils_/729480", "title"=>"SFN pre-incubation increases the GSH/GSSG ratio in primary neutrophils.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-06-24 02:02:16"}
  • {"files"=>["https://ndownloader.figshare.com/files/1098515"], "description"=>"<div><p>The production of high levels of reactive oxygen species by neutrophils is associated with the local and systemic destructive phenotype found in the chronic inflammatory disease periodontitis. In the present study, we investigated the ability of sulforaphane (SFN) to restore cellular glutathione levels and reduce the hyperactivity of circulating neutrophils associated with chronic periodontitis. Using differentiated HL60 cells as a neutrophil model, here we show that generation of extracellular O<sub>2</sub><sup>. -</sup> by the nicotinamide adenine dinucleotide (NADPH) oxidase complex is increased by intracellular glutathione depletion. This may be attributed to the upregulation of thiol regulated acid sphingomyelinase driven lipid raft formation. Intracellular glutathione was also lower in primary neutrophils from periodontitis patients and, consistent with our previous findings, patients neutrophils were hyper-reactive to stimuli. The activity of nuclear factor erythroid-2-related factor 2 (Nrf2), a master regulator of the antioxidant response, is impaired in circulating neutrophils from chronic periodontitis patients. Although patients’ neutrophils exhibit a low reduced glutathione (GSH)/oxidised glutathione (GSSG) ratio and a higher total Nrf2 level, the DNA-binding activity of nuclear Nrf2 remained unchanged relative to healthy controls and had reduced expression of glutamate cysteine ligase catalytic (GCLC), and modifier (GCLM) subunit mRNAs, compared to periodontally healthy subjects neutrophils. Pre-treatment with SFN increased expression of GCLC and GCM, improved intracellular GSH/GSSG ratios and reduced agonist-activated extracellular O<sub>2</sub><sup>. -</sup> production in both dHL60 and primary neutrophils from patients with periodontitis and controls. These findings suggest that a deficiency in Nrf2-dependent pathways may underpin susceptibility to hyper-reactivity in circulating primary neutrophils during chronic periodontitis.</p> </div>", "links"=>[], "tags"=>["sulforaphane", "restores", "cellular", "glutathione", "periodontitis", "neutrophil", "hyperactivity"], "article_id"=>729486, "categories"=>["Biological Sciences"], "users"=>["Irundika H. K. Dias", "Ian L. C. Chapple", "Mike Milward", "Melissa M. Grant", "Eric Hill", "James Brown", "Helen R. Griffiths"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0066407", "stats"=>{"downloads"=>5, "page_views"=>6, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/Sulforaphane_Restores_Cellular_Glutathione_Levels_and_Reduces_Chronic_Periodontitis_Neutrophil_Hyperactivity_In_Vitro_/729486", "title"=>"Sulforaphane Restores Cellular Glutathione Levels and Reduces Chronic Periodontitis Neutrophil Hyperactivity <i>In Vitro</i>", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2013-06-24 02:02:16"}
  • {"files"=>["https://ndownloader.figshare.com/files/1098512"], "description"=>"<p>(A) Western blotting (left) and quantification (right) of total Nrf2 expression in the presence or absence of SFN treatment. (B) Western blotting (left) and quantification (right) of expression of Keap-1, Values are shown normalised to β-actin. (C) Analysis of Nrf2-DNA binding activity by trans-AM nuclear activity assay. A representative experiment of the three is shown. *** P< 0.01. (D) qPCR analysis of GCLC and GCLM gene expression levels of neutrophil mRNA from patients compared to control. (E) qPCR analysis of GCLC, HMOX1, GCLM and NQO1 gene expression levels in neutrophils treated with 5 µM SFN for 16 hours, values normalised to actin.</p>", "links"=>[], "tags"=>["sfn", "nrf2"], "article_id"=>729483, "categories"=>["Biological Sciences"], "users"=>["Irundika H. K. Dias", "Ian L. C. Chapple", "Mike Milward", "Melissa M. Grant", "Eric Hill", "James Brown", "Helen R. Griffiths"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0066407.g005", "stats"=>{"downloads"=>1, "page_views"=>5, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Effect_of_SFN_on_Nrf2_protein_expression_and_activity_in_primary_neutrophils_/729483", "title"=>"Effect of SFN on Nrf2 protein expression and activity in primary neutrophils.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-06-24 02:02:16"}
  • {"files"=>["https://ndownloader.figshare.com/files/1098504"], "description"=>"<p>dHL60 cells were treated with 10µM BSO or 5µM SFN for 16 hours. GSH (A) and GSSG (B) concentrations were measured by the DTNB recycling assay in order to determine the GSH/GSSG ratio (C). Mean peak lucigenin chemiluminescence generated by dHL60 cells was plotted (RLU ± standard error of the mean) (D). Data represent three independent experiments of three replicates. Significant differences were calculated with one way ANOVA followed by Tukey’s multiple comparison test, where *P<0.05.</p>", "links"=>[], "tags"=>["bso", "sfn", "pre-incubation", "dhl60", "respiratory"], "article_id"=>729477, "categories"=>["Biological Sciences"], "users"=>["Irundika H. K. Dias", "Ian L. C. Chapple", "Mike Milward", "Melissa M. Grant", "Eric Hill", "James Brown", "Helen R. Griffiths"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0066407.g001", "stats"=>{"downloads"=>2, "page_views"=>7, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Effects_of_BSO_and_SFN_pre_incubation_on_dHL60_cell_GSH_GSSG_ratio_and_respiratory_burst_/729477", "title"=>"Effects of BSO and SFN pre-incubation on dHL60 cell GSH/GSSG ratio and respiratory burst.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-06-24 02:02:16"}

PMC Usage Stats | Further Information

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Relative Metric

{"start_date"=>"2013-01-01T00:00:00Z", "end_date"=>"2013-12-31T00:00:00Z", "subject_areas"=>[{"subject_area"=>"/Biology and life sciences/Biochemistry", "average_usage"=>[266, 468, 593, 703, 804, 903, 993, 1084, 1171, 1256, 1339, 1422, 1492]}, {"subject_area"=>"/Biology and life sciences/Genetics", "average_usage"=>[284, 491, 620, 738, 843, 945, 1043, 1137, 1225, 1315, 1400, 1479, 1555]}, {"subject_area"=>"/Biology and life sciences/Immunology", "average_usage"=>[266, 466, 591, 701, 799, 887, 982, 1067, 1155, 1237, 1317, 1395, 1458]}, {"subject_area"=>"/Medicine and health sciences/Oral medicine", "average_usage"=>[250, 418, 582, 722, 816, 910, 1002, 1101, 1205, 1279, 1324, 1365, 1411, 1447]}, {"subject_area"=>"/Physical sciences/Chemistry", "average_usage"=>[247, 429, 544, 647, 747, 842, 929, 1012, 1099, 1179, 1263, 1339, 1409]}]}
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