Gabriela Nunes Marsiglio-Librais, Eloisa Aparecida Vilas-Boas, Christopher Carlein, Markus Daniel Alexander Hoffmann, Leticia Prates Roma, Angelo Rafael Carpinelli
{"title":"胰腺β细胞中GPR40激活NADPH氧化酶的证据。","authors":"Gabriela Nunes Marsiglio-Librais, Eloisa Aparecida Vilas-Boas, Christopher Carlein, Markus Daniel Alexander Hoffmann, Leticia Prates Roma, Angelo Rafael Carpinelli","doi":"10.1080/13510002.2020.1757877","DOIUrl":null,"url":null,"abstract":"<p><p><b>Objective:</b> Investigate the involvement of the fatty acids receptor GPR40 in the assembly and activation of NADPH oxidase and the implications on pancreatic β-cell function.<b>Methods:</b> BRIN-BD11 β-cells were exposed to GPR40 agonist (GW9508) or linoleic acid in different glucose concentrations. Superoxide and H<sub>2</sub>O<sub>2</sub> were analyzed, respectively, by DHE fluorescence and by fluorescence of the H<sub>2</sub>O<sub>2</sub> sensor, roGFP2-Orp1. Protein contents of p47<sup>phox</sup> in plasma membrane and cytosol were analyzed by western blot. NADPH oxidase role was evaluated by p22<sup>phox</sup> siRNA or by pharmacological inhibition with VAS2870. NOX2 KO islets were used to measure total cytosolic calcium and insulin secretion.<b>Results:</b> GW9508 and linoleic acid increased superoxide and H<sub>2</sub>O<sub>2</sub> contents at 5.6 and 8.3 mM of glucose. In addition, in 5.6 mM, but not at 16.7 mM of glucose, activation of GPR40 led to the translocation of p47<sup>phox</sup> to the plasma membrane. Knockdown of p22<sup>phox</sup> abolished the increase in superoxide after GW9508 and linoleic acid. No differences in insulin secretion were found between wild type and NOX2 KO islets treated with GW9508 or linoleic acid.<b>Discussion:</b> We report for the first time that acute activation of GPR40 leads to NADPH oxidase activation in pancreatic β-cells, without impact on insulin secretion.</p>","PeriodicalId":21096,"journal":{"name":"Redox Report","volume":null,"pages":null},"PeriodicalIF":5.2000,"publicationDate":"2020-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1080/13510002.2020.1757877","citationCount":"6","resultStr":"{\"title\":\"Evidence for NADPH oxidase activation by GPR40 in pancreatic β-cells.\",\"authors\":\"Gabriela Nunes Marsiglio-Librais, Eloisa Aparecida Vilas-Boas, Christopher Carlein, Markus Daniel Alexander Hoffmann, Leticia Prates Roma, Angelo Rafael Carpinelli\",\"doi\":\"10.1080/13510002.2020.1757877\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p><b>Objective:</b> Investigate the involvement of the fatty acids receptor GPR40 in the assembly and activation of NADPH oxidase and the implications on pancreatic β-cell function.<b>Methods:</b> BRIN-BD11 β-cells were exposed to GPR40 agonist (GW9508) or linoleic acid in different glucose concentrations. Superoxide and H<sub>2</sub>O<sub>2</sub> were analyzed, respectively, by DHE fluorescence and by fluorescence of the H<sub>2</sub>O<sub>2</sub> sensor, roGFP2-Orp1. Protein contents of p47<sup>phox</sup> in plasma membrane and cytosol were analyzed by western blot. NADPH oxidase role was evaluated by p22<sup>phox</sup> siRNA or by pharmacological inhibition with VAS2870. NOX2 KO islets were used to measure total cytosolic calcium and insulin secretion.<b>Results:</b> GW9508 and linoleic acid increased superoxide and H<sub>2</sub>O<sub>2</sub> contents at 5.6 and 8.3 mM of glucose. In addition, in 5.6 mM, but not at 16.7 mM of glucose, activation of GPR40 led to the translocation of p47<sup>phox</sup> to the plasma membrane. Knockdown of p22<sup>phox</sup> abolished the increase in superoxide after GW9508 and linoleic acid. No differences in insulin secretion were found between wild type and NOX2 KO islets treated with GW9508 or linoleic acid.<b>Discussion:</b> We report for the first time that acute activation of GPR40 leads to NADPH oxidase activation in pancreatic β-cells, without impact on insulin secretion.</p>\",\"PeriodicalId\":21096,\"journal\":{\"name\":\"Redox Report\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":5.2000,\"publicationDate\":\"2020-12-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://sci-hub-pdf.com/10.1080/13510002.2020.1757877\",\"citationCount\":\"6\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Redox Report\",\"FirstCategoryId\":\"99\",\"ListUrlMain\":\"https://doi.org/10.1080/13510002.2020.1757877\",\"RegionNum\":2,\"RegionCategory\":\"生物学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q1\",\"JCRName\":\"BIOCHEMISTRY & MOLECULAR BIOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Redox Report","FirstCategoryId":"99","ListUrlMain":"https://doi.org/10.1080/13510002.2020.1757877","RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"BIOCHEMISTRY & MOLECULAR BIOLOGY","Score":null,"Total":0}
Evidence for NADPH oxidase activation by GPR40 in pancreatic β-cells.
Objective: Investigate the involvement of the fatty acids receptor GPR40 in the assembly and activation of NADPH oxidase and the implications on pancreatic β-cell function.Methods: BRIN-BD11 β-cells were exposed to GPR40 agonist (GW9508) or linoleic acid in different glucose concentrations. Superoxide and H2O2 were analyzed, respectively, by DHE fluorescence and by fluorescence of the H2O2 sensor, roGFP2-Orp1. Protein contents of p47phox in plasma membrane and cytosol were analyzed by western blot. NADPH oxidase role was evaluated by p22phox siRNA or by pharmacological inhibition with VAS2870. NOX2 KO islets were used to measure total cytosolic calcium and insulin secretion.Results: GW9508 and linoleic acid increased superoxide and H2O2 contents at 5.6 and 8.3 mM of glucose. In addition, in 5.6 mM, but not at 16.7 mM of glucose, activation of GPR40 led to the translocation of p47phox to the plasma membrane. Knockdown of p22phox abolished the increase in superoxide after GW9508 and linoleic acid. No differences in insulin secretion were found between wild type and NOX2 KO islets treated with GW9508 or linoleic acid.Discussion: We report for the first time that acute activation of GPR40 leads to NADPH oxidase activation in pancreatic β-cells, without impact on insulin secretion.
期刊介绍:
Redox Report is a multidisciplinary peer-reviewed open access journal focusing on the role of free radicals, oxidative stress, activated oxygen, perioxidative and redox processes, primarily in the human environment and human pathology. Relevant papers on the animal and plant environment, biology and pathology will also be included.
While emphasis is placed upon methodological and intellectual advances underpinned by new data, the journal offers scope for review, hypotheses, critiques and other forms of discussion.