NADPH oxidase-2 does not contribute to beta-cell glucotoxicity in cultured pancreatic islets from C57BL/6J mice

Author(s)

de Souza, Arnaldo H. ; Santos, Laila R. B. ; Roma, Leticia P. ; Bensellam, Mohammed ; Carpinelli, Angelo R. ; Jonas, Jean-Christophe

• Document type: Journal article
• Source: Molecular and Cellular Endocrinology; v. 439, n. C, p. 354-362, JAN 5 2017.
• Web of Science Citations: 10

Abstract

High glucose-induced oxidative stress and increased NADPH oxidase-2 (NOX2) activity may contribute to the progressive decline of the functional beta-cell mass in type 2 diabetes. To test that hypothesis, we characterized, in islets from male NOX2 knockout (NOX2-KO) and wild-type (WT) C57BL/6J mice cultured for up to 3 weeks at 10 or 30 mmol/l glucose (G10 or G30), the in vitro effects of glucose on cytosolic oxidative stress using probes sensing glutathione oxidation (GRX1-roGFP2), thiol oxidation (roGFP1) or H2O2 (roGFP2-Orp1), on beta-cell stimulus-secretion coupling events and on beta-cell apoptosis. After 1-2 days of culture in G10, the glucose stimulation of insulin secretion (GSIS) was 1.7-fold higher in NOX2-KO vs. WT islets at 20-30 mmol/l glucose despite similar rises in NAD(P)H and intracellular calcium concentration ({[}Ca2+](i)) and no differences in cytosolic GRX1-roGFP2 oxidation. After long-term culture at G10, roGFP1 and roGFP2-Orpl oxidation and beta-cell apoptosis remained low, and the glucose-induced rises in NAD(P)H, {[}Ca2+](i) and GSIS were similarly preserved in both islet types. After prolonged culture at G30, roGFP1 and roGFP2-Orpl oxidation increased in parallel with beta-cell apoptosis, the glucose sensitivity of the NADPH, {[}Ca2+](i) and insulin secretion responses increased, the maximal {[}Ca2+](i) response decreased, but maximal GSIS was preserved. These responses were almost identical in both islet types. In conclusion, NOX2 is a negative regulator of maximal GSIS in C57BL/6J mouse islets, but it does not detectably contribute to the in vitro glucotoxic induction of cytosolic oxidative stress and alterations of beta-cell survival and function. (C) 2016 Elsevier Ireland Ltd. All rights reserved. (AU)

• FAPESP's process: 13/18232-5 - Role of NADPH oxidase on beta-cell function and survival after prolonged culture with different glucose concentrations
• Grantee: Arnaldo Henrique de Souza
• Support type: Scholarships abroad - Research Internship - Doctorate
• FAPESP's process: 11/21299-9 - EFFECTS OF HYPERGLYCEMIA AND HYPERLIPIDEMIA ON INSULIN SECRETION AND NAD(P)H OXIDASE ACTIVITY IN PANCREATIC ISLETS
• Grantee: Arnaldo Henrique de Souza
• Support type: Scholarships in Brazil - Doctorate