胰腺β细胞中GPR40激活NADPH氧化酶的证据。

IF 5.2 2区 生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY Redox Report Pub Date : 2020-12-01 DOI:10.1080/13510002.2020.1757877
Gabriela Nunes Marsiglio-Librais, Eloisa Aparecida Vilas-Boas, Christopher Carlein, Markus Daniel Alexander Hoffmann, Leticia Prates Roma, Angelo Rafael Carpinelli
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引用次数: 6

摘要

目的:探讨脂肪酸受体GPR40参与NADPH氧化酶的组装和激活及其对胰腺β细胞功能的影响。方法:将BRIN-BD11 β-细胞暴露于不同葡萄糖浓度的GPR40激动剂(GW9508)或亚油酸中。通过DHE荧光和H2O2传感器roGFP2-Orp1荧光分别分析超氧化物和H2O2。western blot检测质膜和细胞质中p47phox蛋白的含量。通过p22phox siRNA或VAS2870的药理抑制来评价NADPH氧化酶的作用。用NOX2 - KO胰岛测定总胞质钙和胰岛素分泌。结果:GW9508和亚油酸使5.6和8.3 mM葡萄糖超氧化物和H2O2含量增加。此外,在5.6 mM,而不是16.7 mM的葡萄糖中,GPR40的激活导致p47phox转运到质膜。p22phox的敲除消除了GW9508和亚油酸后超氧化物的增加。GW9508和亚油酸对野生型和NOX2型KO胰岛胰岛素分泌无显著影响。讨论:我们首次报道了GPR40的急性激活导致胰腺β细胞中NADPH氧化酶的激活,而不影响胰岛素分泌。
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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.

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来源期刊
Redox Report
Redox Report 生物-生化与分子生物学
CiteScore
6.10
自引率
0.00%
发文量
28
审稿时长
>12 weeks
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.
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