OBJECTIVE: By exposing mice carrying a deletion of NADPH oxidase isoform 4, NOX4, specifically in pancreatic β cells (βNOX4-/-) to nutrient excess stimulated by a high-fat diet (HFD), this study aimed to elucidate the role of β-cell redox status in the development of meta-inflammation within the diabetic phenotype. METHODS: The authors performed basic phenotyping of βNOX4-/- mice on HFD involving insulin and glycemic analyses, histochemistry of adipocytes, indirect calorimetry, and cytokine analyses. To characterize local inflammation, the study used caspase-1 activity assay, interleukin-1β immunochemistry, and real-time polymerase chain reaction during coculturing of β cells with macrophages. RESULTS: The phenotype of βNOX4-/- mice on HFD was not associated with hyperinsulinemia and hyperglycemia but showed accumulation of excessive lipids in epididymal fat and β cells. Surprisingly, mice showed significantly reduced systemic inflammation. Decreased interleukin-1β protein levels and downregulated NLRP3-inflammasome activity were observed on chronic glucose overload in βNOX4-/- isolated islets and NOX4-silenced INS1-E cells resulting in attenuated proinflammatory polarization of macrophages/monocytes in vitro and in situ and reduced local islet inflammation. CONCLUSIONS: Experimental evidence suggests that NOX4 pro-oxidant activity in β cells is involved in NLRP3-inflammasome activation during chronic nutrient overload and participates in local inflammatory signaling and perhaps toward peripheral tissues, contributing to a diabetic inflammatory phenotype.

1st Faculty of Medicine Charles University Prague Czech Republic

Department of Medical Chemistry and Biochemistry Palacký University Olomouc Czech Republic

Faculty of Science Charles University Prague Czech Republic

Laboratory of Mitochondrial Physiology Institute of Physiology of the Czech Academy of Sciences Prague Czech Republic

Laboratory of Pancreatic Islet Research Institute of Physiology of the Czech Academy of Sciences Prague Czech Republic

Obesity (Silver Spring). 2024 Jul;32(7):1410. doi: 10.1002/oby.24045. PubMed

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Ahmed Alfar E, Kirova D, Konantz J, Birke S, Mansfeld J, Ninov N. Distinct levels of reactive oxygen species coordinate metabolic activity with beta-cell mass plasticity. Sci Rep. 2017;7:3994.

Swisa A, Glaser B, Dor Y. Metabolic stress and compromised identity of pancreatic beta cells. Front Genet. 2017;8:21.

Jezek P, Holendova B, Plecita-Hlavata L. Redox signaling from mitochondria: signal propagation and its targets. Biomolecules. 2020;10:93.

Stancill JS, Broniowska KA, Oleson BJ, Naatz A, Corbett JA. Pancreatic beta-cells detoxify H2O2 through the peroxiredoxin/thioredoxin antioxidant system. J Biol Chem. 2019;294:4843-4853.

Plecita-Hlavata L, Jaburek M, Holendova B, et al. Glucose-stimulated insulin secretion fundamentally requires H2O2 signaling by NADPH oxidase 4. Diabetes. 2020;69:1341-1354.

Plecita-Hlavata L, Engstova H, Holendova B, et al. Mitochondrial superoxide production decreases on glucose-stimulated insulin secretion in pancreatic beta cells due to decreasing mitochondrial matrix NADH/NAD(+) ratio. Antioxid Redox Signal. 2020;33:789-815.

Zhang L, Keung W, Samokhvalov V, Wang W, Lopaschuk GD. Role of fatty acid uptake and fatty acid beta-oxidation in mediating insulin resistance in heart and skeletal muscle. Biochim Biophys Acta. 2010;1801:1-22.

Jezek P, Jaburek M, Plecita-Hlavata L. Contribution of oxidative stress and impaired biogenesis of pancreatic beta-cells to type 2 diabetes. Antioxid Redox Signal. 2019;31:722-751.

Vilas-Boas EA, Nalbach L, Ampofo E, et al. Transient NADPH oxidase 2-dependent H2O2 production drives early palmitate-induced lipotoxicity in pancreatic islets. Free Radic Biol Med. 2021;162:1-13.

Li N, Li B, Brun T, et al. NADPH oxidase NOX2 defines a new antagonistic role for reactive oxygen species and cAMP/PKA in the regulation of insulin secretion. Diabetes. 2012;61:2842-2850.

Surwit RS, Kuhn CM, Cochrane C, McCubbin JA, Feinglos MN. Diet-induced type II diabetes in C57BL/6J mice. Diabetes. 1988;37:1163-1167.

Lumeng CN, Saltiel AR. Inflammatory links between obesity and metabolic disease. J Clin Invest. 2011;121:2111-2117.

Tiedge M, Lortz S, Drinkgern J, Lenzen S. Relation between antioxidant enzyme gene expression and antioxidative defense status of insulin-producing cells. Diabetes. 1997;46:1733-1742.

Heydemann A. An overview of murine high fat diet as a model for type 2 diabetes mellitus. J Diabetes Res. 2016;2016:2902351.

Zhang Y, Chen F. Reactive oxygen species (ROS), troublemakers between nuclear factor-kappaB (NF-kappaB) and c-Jun NH(2)-terminal kinase (JNK). Cancer Res. 2004;64:1902-1905.

Benakova S, Holendova B, Plecita-Hlavata L. Redox homeostasis in pancreatic beta-cells: from development to failure. Antioxidants (Basel). 2021;10:10.

Collier JJ, Sparer TE, Karlstad MD, Burke SJ. Pancreatic islet inflammation: an emerging role for chemokines. J Mol Endocrinol. 2017;59:R33-R46.

Eguchi K, Manabe I, Oishi-Tanaka Y, et al. Saturated fatty acid and TLR signaling link beta cell dysfunction and islet inflammation. Cell Metab. 2012;15:518-533.

Dhingra S, Sharma AK, Singla DK, Singal PK. p38 and ERK1/2 MAPKs mediate the interplay of TNF-alpha and IL-10 in regulating oxidative stress and cardiac myocyte apoptosis. Am J Physiol Heart Circ Physiol. 2007;293:H3524-H3531.

Zhou R, Tardivel A, Thorens B, Choi I, Tschopp J. Thioredoxin-interacting protein links oxidative stress to inflammasome activation. Nat Immunol. 2010;11:136-140.

Li Y, Mouche S, Sajic T, et al. Deficiency in the NADPH oxidase 4 predisposes towards diet-induced obesity. Int J Obes (Lond). 2012;36:1503-1513.

Anvari E, Wikstrom P, Walum E, Welsh N. The novel NADPH oxidase 4 inhibitor GLX351322 counteracts glucose intolerance in high-fat diet-treated C57BL/6 mice. Free Radic Res. 2015;49:1308-1318.

Moon JS, Nakahira K, Chung KP, et al. NOX4-dependent fatty acid oxidation promotes NLRP3 inflammasome activation in macrophages. Nat Med. 2016;22:1002-1012.

Sireesh D, Dhamodharan U, Ezhilarasi K, Vijay V, Ramkumar KM. Association of NF-E2 related factor 2 (Nrf2) and inflammatory cytokines in recent onset type 2 diabetes mellitus. Sci Rep. 2018;8:5126.

Barter PJ, Nicholls S, Rye KA, Anantharamaiah GM, Navab M, Fogelman AM. Antiinflammatory properties of HDL. Circ Res. 2004;95:764-772.

Boni-Schnetzler M, Thorne J, Parnaud G, et al. Increased interleukin (IL)-1beta messenger ribonucleic acid expression in beta-cells of individuals with type 2 diabetes and regulation of IL-1beta in human islets by glucose and autostimulation. J Clin Endocrinol Metab. 2008;93:4065-4074.

Lewis GF, Carpentier A, Adeli K, Giacca A. Disordered fat storage and mobilization in the pathogenesis of insulin resistance and type 2 diabetes. Endocr Rev. 2002;23:201-229.

Sae-Lee C, Moolsuwan K, Chan L, Poungvarin N. ChREBP regulates itself and metabolic genes implicated in lipid accumulation in beta-cell line. PLoS One. 2016;11:e0147411.

Eto K, Yamashita T, Matsui J, Terauchi Y, Noda M, Kadowaki T. Genetic manipulations of fatty acid metabolism in beta-cells are associated with dysregulated insulin secretion. Diabetes. 2002;51(Suppl 3):S414-S420.

Wicksteed B, Brissova M, Yan W, et al. Conditional gene targeting in mouse pancreatic ß-cells: analysis of ectopic Cre transgene expression in the brain. Diabetes. 2010;59:3090-3098.

Vallet P, Charnay Y, Steger K, et al. Neuronal expression of the NADPH oxidase NOX4, and its regulation in mouse experimental brain ischemia. Neuroscience. 2005;132:233-238.

Nayernia Z, Jaquet V, Krause KH. New insights on NOX enzymes in the central nervous system. Antioxid Redox Signal. 2014;20:2815-2837.

Steel DM, Whitehead AS. The major acute phase reactants: C-reactive protein, serum amyloid P component and serum amyloid a protein. Immunol Today. 1994;15:81-88.

De Nardo D, Labzin LI, Kono H, et al. High-density lipoprotein mediates anti-inflammatory reprogramming of macrophages via the transcriptional regulator ATF3. Nat Immunol. 2014;15:152-160.

Rider P, Carmi Y, Guttman O, et al. IL-1alpha and IL-1beta recruit different myeloid cells and promote different stages of sterile inflammation. J Immunol. 2011;187:4835-4843.

Boni-Schnetzler M, Boller S, Debray S, et al. Free fatty acids induce a proinflammatory response in islets via the abundantly expressed interleukin-1 receptor I. Endocrinology. 2009;150:5218-5229.

Nordmann TM, Dror E, Schulze F, et al. The role of inflammation in beta-cell dedifferentiation. Sci Rep. 2017;7:6285.

Dror E, Dalmas E, Meier DT, et al. Postprandial macrophage-derived IL-1beta stimulates insulin, and both synergistically promote glucose disposal and inflammation. Nat Immunol. 2017;18:283-292.

Ying W, Lee YS, Dong Y, et al. Expansion of islet-resident macrophages leads to inflammation affecting beta cell proliferation and function in obesity. Cell Metab. 2019;29(2):457-474.e5.

Hajmrle C, Smith N, Spigelman AF, et al. Interleukin-1 signaling contributes to acute islet compensation. JCI Insight. 2016;1:e86055.

Salama A, Fichou N, Allard M, et al. MicroRNA-29b modulates innate and antigen-specific immune responses in mouse models of autoimmunity. PLoS One. 2014;9:e106153.

Cianciaruso C, Phelps EA, Pasquier M, et al. Primary human and rat beta-cells release the intracellular autoantigens GAD65, IA-2, and proinsulin in exosomes together with cytokine-induced enhancers of immunity. Diabetes. 2017;66:460-473.

Weitz JR, Makhmutova M, Almaca J, et al. Mouse pancreatic islet macrophages use locally released ATP to monitor beta cell activity. Diabetologia. 2018;61:182-192.

Jarc E, Petan T. A twist of FATe: lipid droplets and inflammatory lipid mediators. Biochimie. 2020;169:69-87.

Redox Status as a Key Driver of Healthy Pancreatic Beta-Cells