American Diabetes Association
REVISION_SUPPLEMENTAL DATA Plecita et al &IR version DIABETES Jezek corresponding.docx (6.5 MB)

Glucose-stimulated insulin secretion fundamentally requires H2O2 signaling by NADPH oxidase 4

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Version 2 2020-05-14, 20:27
Version 1 2020-04-03, 22:51
posted on 2020-05-14, 20:27 authored by Ada AdminAda Admin, Lydie Plecitá-Hlavatá, Martin Jabůrek, Blanka Holendová, Jan Tauber, Vojtěch Pavluch, Zuzana Berková, Monika Cahová, Katrin Schroeder, Ralf P. Brandes, Detlef Siemen, Petr Ježek
NADPH facilitates glucose-stimulated insulin secretion (GSIS) in pancreatic islet (PI) b-cells by an as yet unknown mechanism. We found NADPH oxidase, isoform-4 (NOX4), to be the major producer of cytosolic H2O2, essential for GSIS, while the increase in ATP/ADP alone was insufficient. The fast GSIS phase was absent in PIs from NOX4-null, b-cell-specific knockout mice (NOX4bKO) (not NOX2KO), and NOX4-silenced or catalase-overexpressing INS-1E cells. Lentiviral NOX4 overexpression or H2O2 rescued GSIS in PIs from NOX4bKO mice. NOX4 silencing suppressed Ca2+ oscillations and the patch-clamped ATP-sensitive potassium channel (KATP) opened more frequently at high glucose. Mitochondrial H2O2, decreasing upon GSIS, provided an alternative redox signaling when 2-oxo-isocaproate or fatty acid oxidation formed superoxide by electron-transport flavoprotein:Q-oxidoreductase. Unlike GSIS, this ceased with mitochondrial antioxidant SkQ1. Both NOX4KO and NOX4bKO strains exhibited impaired glucose tolerance and peripheral insulin resistance. Thus the redox signaling previously suggested to cause b-cell-self-checking – hypothetically induces insulin resistance when absent. In conclusion, ATP plus H2O2 elevations constitute an essential switch-on signal of insulin exocytosis for glucose and branched-chain oxoacids as secretagogues (partly for fatty acids). Redox signaling could be impaired by cytosolic antioxidants, hence those targeting mitochondria should be preferred for clinical applications to treat (pre)diabetes at any stage.


The project was supported by grants from the Grant Agency of the Czech Republic (No. 16-06700S to L.P.H. and 20-00408S plus 17-01813S to P.J.), plus by institutional support RVO:67985823.