Possible Protective Effect of Membrane Lipid Rafts against Interleukin-1β-Mediated Anti-Proliferative Effect in INS-1 Cells
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{"title"=>"Possible protective effect of membrane lipid rafts against interleukin-1β-mediated anti-proliferative effect in INS-1 cells", "type"=>"journal", "authors"=>[{"first_name"=>"Myriam", "last_name"=>"Chentouf", "scopus_author_id"=>"12767368800"}, {"first_name"=>"Caroline", "last_name"=>"Guzman", "scopus_author_id"=>"55649964700"}, {"first_name"=>"Moustafa", "last_name"=>"Hamze", "scopus_author_id"=>"55744365400"}, {"first_name"=>"René", "last_name"=>"Gross", "scopus_author_id"=>"7403106641"}, {"first_name"=>"Anne Dominique", "last_name"=>"Lajoix", "scopus_author_id"=>"6602711155"}, {"first_name"=>"Sylvie", "last_name"=>"Peraldi-Roux", "scopus_author_id"=>"6701544000"}], "year"=>2014, "source"=>"PLoS ONE", "identifiers"=>{"pmid"=>"25068701", "sgr"=>"84904878298", "doi"=>"10.1371/journal.pone.0102889", "scopus"=>"2-s2.0-84904878298", "pui"=>"373631633", "isbn"=>"1932-6203 (Electronic)\r1932-6203 (Linking)", "issn"=>"19326203"}, "id"=>"1255dc05-51f6-3fdb-9837-4703c88c07c5", "abstract"=>"We recently reported that pancreatic islets from pre-diabetic rats undergo an inflammatory process in which IL-1β takes part and controls β-cell function. In the present study, using the INS-1 rat pancreatic β-cell line, we investigated the potential involvement of membrane-associated cholesterol-enriched lipid rafts in IL-1β signaling and biological effects on insulin secretion, β-cell proliferation and apoptosis. We show that, INS-1 cells exposure to increasing concentrations of IL-1β leads to a progressive inhibition of insulin release, an increase in the number of apoptotic cells and a dose-dependent decrease in pancreatic β-cell proliferation. Disruption of membrane lipid rafts markedly reduced glucose-stimulated insulin secretion but did not affect either cell apoptosis or proliferation rate, demonstrating that membrane lipid raft integrity is essential for β-cell secretory function. In the same conditions, IL-1β treatment of INS-1 cells led to a slight further decrease in insulin secretion for low concentrations of the cytokine, and a more marked one, similar to that observed in normal cells for higher concentrations. These effects occurred together with an increase in iNOS expression and surprisingly with an upregulation of tryptophane hydroxylase and protein Kinase C in membrane lipid rafts suggesting that compensatory mechanisms develop to counteract IL-1β inhibitory effects. We also demonstrate that disruption of membrane lipid rafts did not prevent cytokine-induced cell death recorded after exposure to high IL-1β concentrations. Finally, concerning cell proliferation, we bring strong evidence that membrane lipid rafts exert a protective effect against IL-1β anti-proliferative effect, possibly mediated at least partly by modifications in ERK and PKB expression/activities. Our results 1) demonstrate that IL-1β deleterious effects do not require a cholesterol-dependent plasma membrane compartmentalization of IL-1R1 signaling and 2) confer to membrane lipid rafts integrity a possible protective function that deserves to be considered in the context of inflammation and especially T2D pathogenesis.", "link"=>"http://www.mendeley.com/research/possible-protective-effect-membrane-lipid-rafts-against-interleukin1%CE%B2mediated-antiproliferative-effe", "reader_count"=>7, "reader_count_by_academic_status"=>{"Researcher"=>2, "Student > Ph. D. Student"=>2, "Student > Postgraduate"=>1, "Student > Bachelor"=>1, "Lecturer"=>1}, "reader_count_by_user_role"=>{"Researcher"=>2, "Student > Ph. D. Student"=>2, "Student > Postgraduate"=>1, "Student > Bachelor"=>1, "Lecturer"=>1}, "reader_count_by_subject_area"=>{"Biochemistry, Genetics and Molecular Biology"=>2, "Agricultural and Biological Sciences"=>3, "Medicine and Dentistry"=>2}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>2}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>3}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>2}}, "group_count"=>1}

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/1611944"], "description"=>"<p>(A) Immunofluorescence studies in the INS-1 cell line; Immunostaining of receptor IL1-R1 and cytokine IL1-β. Irrelevant rabbit IgG and secondary antibody were used as negative controls. (B) Characterization of lipid raft fractions by Ganglioside M1 detection after isolation on sucrose gradient; Dot Blot characterization of Brij 98-extracted fractions. INS-1 cells were lysed with Brij 98 detergent at 37°C and separated into fractions by sucrose density gradient. Membrane rafts are present in fractions 3 and 4, as indicated by GM1 expression. Membrane lipid rafts localization in the INS-1 cell line by GM1 detection using immunofluorescence, without and with MβCD treatment. Results are representative of three independent experiments. (C) Western blots of IL-1R1 expression in lipid-rafts and non-rafts fractions. Forty micrograms of protein extracts from membrane lipid rafts- and non-lipid rafts were run on SDS-PAGE and blotted with anti Il-1R1 antibody.</p>", "links"=>[], "tags"=>["cell biology", "Signal transduction", "cell signaling", "physiology", "Immune physiology", "cytokines", "Metabolic disorders", "Diabetes mellitus", "type 2 diabetes", "expresses", "il-1r1", "lipid"], "article_id"=>1120200, "categories"=>["Biological Sciences"], "users"=>["Myriam Chentouf", "Caroline Guzman", "Moustafa Hamze", "René Gross", "Anne Dominique Lajoix", "Sylvie Peraldi-Roux"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0102889.g001", "stats"=>{"downloads"=>1, "page_views"=>18, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_INS_1_cell_line_expresses_IL_1R1_in_lipid_rafts_/1120200", "title"=>"INS-1 cell line expresses IL-1R1 in lipid rafts.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-07-28 03:07:06"}
  • {"files"=>["https://ndownloader.figshare.com/files/1611952"], "description"=>"<p>Changes in the expression pattern of proteins involved in signal transduction, apoptosis, and cell cycle were examined by antibody Array. Protein extracts (1 mg/ml) from INS-1 cells membrane lipid rafts and non lipid rafts, treated or not by IL1-β, were labeled with Cy3 and Cy5. Equal amounts of labelled proteins were incubated with antibodies doted on nitrocellulose-coated glass slides. The level of protein expression was quantified and the most representative changes are summarized on this Table.</p><p>Up or down regulated membrane lipid raft and non lipid raft proteins after IL-1β treatment are presented in relative values recorded for the same proteins expressed in raft and non raft fraction in control condition (no IL-1β treatment). A 2-fold increase or decrease (bold) taken as cut-off value (positive or negative) is considered as significant.</p>", "links"=>[], "tags"=>["cell biology", "Signal transduction", "cell signaling", "physiology", "Immune physiology", "cytokines", "Metabolic disorders", "Diabetes mellitus", "type 2 diabetes", "lipid", "raft-", "non-lipid", "raft"], "article_id"=>1120208, "categories"=>["Biological Sciences"], "users"=>["Myriam Chentouf", "Caroline Guzman", "Moustafa Hamze", "René Gross", "Anne Dominique Lajoix", "Sylvie Peraldi-Roux"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0102889.t001", "stats"=>{"downloads"=>0, "page_views"=>4, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_IL1_946_effects_on_protein_expression_in_lipid_raft_and_non_lipid_raft_fractions_/1120208", "title"=>"IL1-β effects on protein expression in lipid raft- and non-lipid raft fractions.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2014-07-28 03:07:06"}
  • {"files"=>["https://ndownloader.figshare.com/files/1611950"], "description"=>"<p>(A) Insulin secretion assay on INS-1 cells cultured in the presence of increasing IL-1β concentrations for 2 days. At the end of the exposure time, cells were washed, deprived in glucose during 1 hour and incubated in the presence of 0, 2.8, 5.6, 8.3 and 16 mM glucose. Cells supernatant fractions were collected and insulin content extracted with acid/alcohol mixture. Insulin present in culture supernatants and insulin content was quantified using HTRF assay. (B) Insulin secretion with IL-1β treatment (White column), and after pre-treatment of INS-1 cells with 500 ng/ml of IL1 receptor 1 antagonist (IL1-RA) to inhibit IL1-β effect on secretion (Black column). Insulin has been quantified by HTRF. <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0102889#pone-0102889-g002\" target=\"_blank\">Fig (2A-2B</a>): All data represent insulin release normalized for insulin contents and are expressed as percentages of insulin secretion recorded in the presence of 16 mM glucose alone; they are all the result of 3 independent experiments, with each experimental condition performed in triplicate. (C) Annexin V labeling of apoptotic INS-1 cells after treatment by increased concentrations of IL-1β: Ins-1 cells were analyzed by confocal microscopy (pictures are representative of 3 independent experiments) and flow cytometry (see <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0102889#pone-0102889-g003\" target=\"_blank\">Fig 3B</a>). (D) Cell proliferation test by BrdU incorporation in INS-1 cells after IL-1β treatment; controls are cells not exposed to the cytokine. Data represent means of 3 independent experiments with each experimental condition performed in triplicates. *: p<0.05 and **: p<0.01 vs cells incubated in the absence of IL-1 β; a: p<0.05 and b: p<0.01 vs in the absence of antagonist.</p>", "links"=>[], "tags"=>["cell biology", "Signal transduction", "cell signaling", "physiology", "Immune physiology", "cytokines", "Metabolic disorders", "Diabetes mellitus", "type 2 diabetes", "insulin", "apoptose"], "article_id"=>1120206, "categories"=>["Biological Sciences"], "users"=>["Myriam Chentouf", "Caroline Guzman", "Moustafa Hamze", "René Gross", "Anne Dominique Lajoix", "Sylvie Peraldi-Roux"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0102889.g002", "stats"=>{"downloads"=>0, "page_views"=>2, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Effect_of_IL_1_946_on_946_cell_function_insulin_secretion_apoptose_and_proliferation_/1120206", "title"=>"Effect of IL-1β on β-cell function: insulin secretion, apoptose and proliferation.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-07-28 03:07:06"}
  • {"files"=>["https://ndownloader.figshare.com/files/1611951"], "description"=>"<p>(A) Insulin secretion after MβCD treatment to induce cholesterol depletion and disturb lipid raft domains. INS-1 cells were stimulated by increasing concentrations of IL1-β and insulin secretion was quantified by HTRF assay. INS-1 cell without MβCD treatment (White column), and INS-1 cell pre-treated by MβCD (Black column). All data represent insulin release normalized for insulin contents and are expressed as percentages of insulin secretion recorded in the absence of any treatment. (B) INS-1 apoptosis. INS-1 cells were stimulated by IL-1β with or without MβCD pre-treatment, and induction of apoptosis was determined using Annexin V FITC/propidium iodide staining in flow cytometry. (C) Analysis of INS-1 proliferation using BrdU incorporation, with or without MβCD treatment and after IL-1β stimulation. INS-1 cell without MβCD treatment (White column), and INS-1 cell pre-treated by MβCD (Black column). <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0102889#pone-0102889-g003\" target=\"_blank\">Fig 3</a> (A-B-C) Data are means of 3 experiments with each experimental condition performed as triplicates. *: p<0.05 and **: p<0.01 vs non- IL-1β pretreated cells; a: p<0.05 and b: p<0.01 vs non- MβCD treated cells.</p>", "links"=>[], "tags"=>["cell biology", "Signal transduction", "cell signaling", "physiology", "Immune physiology", "cytokines", "Metabolic disorders", "Diabetes mellitus", "type 2 diabetes", "rafts", "insulin", "apoptosis"], "article_id"=>1120207, "categories"=>["Biological Sciences"], "users"=>["Myriam Chentouf", "Caroline Guzman", "Moustafa Hamze", "René Gross", "Anne Dominique Lajoix", "Sylvie Peraldi-Roux"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0102889.g003", "stats"=>{"downloads"=>1, "page_views"=>5, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Lipid_rafts_integrity_and_IL_1_946_effects_on_insulin_secretion_apoptosis_and_cell_proliferation_/1120207", "title"=>"Lipid rafts integrity and IL-1β effects on insulin secretion, apoptosis and cell proliferation.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-07-28 03:07:06"}

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