Nicotinamide adenine dinucleotide phosphate oxidases (NOX-es) produce reactive oxygen species and modulate β-cell insulin secretion. Islets of type 2 diabetic subjects present elevated expression of NOX5. Here, we sought to characterize regulation of NOX5 expression in human islets in vitro and to uncover the relevance of NOX5 in islet function in vivo using a novel mouse model expressing NOX5 in doxycycline-inducible, β-cell-specific manner (RIP/rtTA/NOX5 mice).
Results:
In situ hybridization and immunohistochemistry employed on pancreatic sections demonstrated NOX5 messenger ribonucleic acid (mRNA) and protein expressions in human islets. In cultures of dispersed islets, NOX5 protein was observed in somatostatin-positive (δ) cells in basal (2.8 mM glucose) conditions. Small interfering ribonucleic acid (siRNA)-mediated knockdown of NOX5 in human islets cultured in basal glucose concentrations resulted in diminished glucose-induced insulin secretion (GIIS) in vitro. However, when islets were preincubated in high (16.7 mM) glucose media for 12 h, NOX5 appeared also in insulin-positive (β) cells. In vivo, mice with β-cell NOX5 expression developed aggravated impairment of GIIS compared with control mice when challenged with 14 weeks of high-fat diet. Similarly, in vitro palmitate preincubation resulted in more severe reduction of insulin release in islets of RIP/rtTA/NOX5 mice compared with their control littermates. Decreased insulin secretion was most distinct in response to theophylline stimulation, suggesting impaired cyclic adenosine monophosphate (cAMP)-mediated signaling due to increased phosphodiesterase activation.
Innovation and Conclusions:
Our data provide the first insight into the complex regulation and function of NOX5 in islets implying an important role for NOX5 in δ-cell-mediated intraislet crosstalk in physiological circumstances but also identifying it as an aggravating factor in β-cell failure in diabetic conditions.
Get full access to this article
View all access options for this article.
References
1.
AntonyS, WuY, HewittSM, AnverMR, ButcherD, JiangG, MeitzlerJL, LiuH, JuhaszA, LuJ, RoyKK, and DoroshowJH. Characterization of NADPH oxidase 5 expression in human tumors and tumor cell lines with a novel mouse monoclonal antibody. Free Radic Biol Med, 65: 497–508, 2013.
2.
AnvariE, WikstromP, WalumE, and WelshN. The novel NADPH oxidase 4 inhibitor GLX351322 counteracts glucose intolerance in high-fat diet-treated C57BL/6 mice. Free Radic Res, 49: 1–11, 2015.
3.
AshcroftSJ and ChristieMR. Effects of glucose on the cytosolic ration of reduced/oxidized nicotinamide-adenine dinucleotide phosphate in rat islets of Langerhans. Biochem J, 184: 697–700, 1979.
4.
BanfiB, MolnarG, MaturanaA, StegerK, HegedusB, DemaurexN, and KrauseKH. A Ca(2+)-activated NADPH oxidase in testis, spleen, and lymph nodes. J Biol Chem, 276: 37594–37601, 2001.
5.
BanfiB, TironeF, DurusselI, KniszJ, MoskwaP, MolnarGZ, KrauseKH, and CoxJA. Mechanism of Ca2+ activation of the NADPH oxidase 5 (NOX5). J Biol Chem, 279: 18583–18591, 2004.
6.
BedardK, JaquetV, and KrauseKH. NOX5: from basic biology to signaling and disease. Free Radic Biol Med, 52: 725–734, 2012.
7.
BelAibaRS, DjordjevicT, PetryA, DiemerK, BonelloS, BanfiB, HessJ, PogrebniakA, BickelC, and GorlachA. NOX5 variants are functionally active in endothelial cells. Free Radic Biol Med, 42: 446–459, 2007.
8.
BjorklundA, LansnerA, and GrillVE. Glucose-induced [Ca2+]i abnormalities in human pancreatic islets: important role of overstimulation. Diabetes, 49: 1840–1848, 2000.
9.
BlodgettDM, NowosielskaA, AfikS, PechholdS, CuraAJ, KennedyNJ, KimS, KucukuralA, DavisRJ, KentSC, GreinerDL, GarberMG, HarlanDM, and diIorioP. Novel observations from next-generation RNA sequencing of highly purified human adult and fetal islet cell subsets. Diabetes, 64: 3172–3181, 2015.
10.
BouzakriK, ZachrissonA, Al-KhaliliL, ZhangBB, KoistinenHA, KrookA, and ZierathJR. siRNA-based gene silencing reveals specialized roles of IRS-1/Akt2 and IRS-2/Akt1 in glucose and lipid metabolism in human skeletal muscle. Cell Metab, 4: 89–96, 2006.
11.
CarnesecchiS, RougemontAL, DoroshowJH, NagyM, MoucheS, Gumy-PauseF, and SzantoI. The NADPH oxidase NOX5 protects against apoptosis in ALK-positive anaplastic large-cell lymphoma cell lines. Free Radic Biol Med, 84: 22–29, 2015.
12.
CasasAI, KleikersPW, GeussE, LanghauserF, AdlerT, BuschDH, Gailus-DurnerV, de AngelisMH, EgeaJ, LopezMG, KleinschnitzC, and SchmidtHH. Calcium-dependent blood-brain barrier breakdown by NOX5 limits postreperfusion benefit in stroke. J Clin Invest, 129: 1772–1778, 2019.
13.
ChenF, HaighS, YuY, BensonT, WangY, LiX, DouH, BagiZ, VerinAD, SteppDW, CsanyiG, ChadliA, WeintraubNL, SmithSM, and FultonDJ. Nox5 stability and superoxide production is regulated by C-terminal binding of Hsp90 and CO-chaperones. Free Radic Biol Med, 89: 793–805, 2015.
14.
ChenF, PandeyD, ChadliA, CatravasJD, ChenT, and FultonDJ. Hsp90 regulates NADPH oxidase activity and is necessary for superoxide but not hydrogen peroxide production. Antioxid Redox Signal, 14: 2107–2119, 2011.
15.
ChenF, YuY, QianJ, WangY, ChengB, DimitropoulouC, PatelV, ChadliA, RudicRD, SteppDW, CatravasJD, and FultonDJ. Opposing actions of heat shock protein 90 and 70 regulate nicotinamide adenine dinucleotide phosphate oxidase stability and reactive oxygen species production. Arterioscler Thromb Vasc Biol, 32: 2989–2999, 2012.
16.
ChengG, CaoZ, XuX, van MeirEG, and LambethJD. Homologs of gp91phox: cloning and tissue expression of Nox3, Nox4, and Nox5. Gene, 269: 131–140, 2001.
17.
Cifuentes-PaganoE, MeijlesDN, and PaganoPJ. The quest for selective nox inhibitors and therapeutics: challenges, triumphs and pitfalls. Antioxid Redox Signal, 20: 2741–2754, 2014.
18.
CostfordSR, Castro-AlvesJ, ChanKL, BaileyLJ, WooM, BelshamDD, BrumellJH, and KlipA. Mice lacking NOX2 are hyperphagic and store fat preferentially in the liver. Am J Physiol Endocrinol Metab, 306: E1341–E1353, 2014.
19.
EvansJL, GoldfineID, MadduxBA, and GrodskyGM. Oxidative stress and stress-activated signaling pathways: a unifying hypothesis of type 2 diabetes. Endocr Rev, 23: 599–622, 2002.
20.
FultonDJ. Nox5 and the regulation of cellular function. Antioxid Redox Signal, 11: 2443–2452, 2009.
21.
GeisztM, KoppJB, VarnaiP, and LetoTL. Identification of renox, an NAD(P)H oxidase in kidney. Proc Natl Acad Sci U S A, 97: 8010–8014, 2000.
22.
GraySM, MeijerRI, and BarrettEJ. Insulin regulates brain function, but how does it get there?. Diabetes, 63: 3992–3997, 2014.
23.
GuichardC, MoreauR, PessayreD, EppersonTK, and KrauseKH. NOX family NADPH oxidases in liver and in pancreatic islets: a role in the metabolic syndrome and diabetes?. Biochem Soc Trans, 36: 920–929, 2008.
24.
GuntonJE, KulkarniRN, YimS, OkadaT, HawthorneWJ, TsengYH, RobersonRS, RicordiC, O'ConnellPJ, GonzalezFJ, and KahnCR. Loss of ARNT/HIF1beta mediates altered gene expression and pancreatic-islet dysfunction in human type 2 diabetes. Cell, 122: 337–349, 2005.
25.
GuzikTJ, ChenW, GongoraMC, GuzikB, LobHE, MangalatD, HochN, DikalovS, RudzinskiP, KapelakB, SadowskiJ, and HarrisonDG. Calcium-dependent NOX5 nicotinamide adenine dinucleotide phosphate oxidase contributes to vascular oxidative stress in human coronary artery disease. J Am Coll Cardiol, 52: 1803–1809, 2008.
26.
Hauge-EvansAC, KingAJ, CarmignacD, RichardsonCC, RobinsonIC, LowMJ, ChristieMR, PersaudSJ, and JonesPM. Somatostatin secreted by islet delta-cells fulfills multiple roles as a paracrine regulator of islet function. Diabetes, 58: 403–411, 2009.
27.
HedeskovCJ, CapitoK, and ThamsP. Cytosolic ratios of free [NADPH]/[NADP+] and [NADH]/[NAD+] in mouse pancreatic islets, and nutrient-induced insulin secretion. Biochem J, 241: 161–167, 1987.
28.
HoltermanCE, ThibodeauJF, TowaijC, GutsolA, MontezanoAC, ParksRJ, CooperME, TouyzRM, and KennedyCR. Nephropathy and elevated BP in mice with podocyte-specific NADPH oxidase 5 expression. J Am Soc Nephrol, 25: 784–797, 2014.
29.
IurlaroR and Munoz-PinedoC. Cell death induced by endoplasmic reticulum stress. FEBS J, 283: 2640–2652, 2016.
30.
IvarssonR, QuintensR, DejongheS, TsukamotoK, in ‘t VeldP, RenstromE, and SchuitFC. Redox control of exocytosis: regulatory role of NADPH, thioredoxin, and glutaredoxin. Diabetes, 54: 2132–2142, 2005.
JohnsonJD. A practical guide to genetic engineering of pancreatic beta-cells in vivo: getting a grip on RIP and MIP. Islets, 6: e944439, 2014.
33.
KleikersPW, DaoVT, GobE, HooijmansC, DebetsJ, van EssenH, KleinschnitzC, and SchmidtHH. SFRR-E Young Investigator AwardeeNOXing out stroke: identification of NOX4 and 5as targets in blood-brain-barrier stabilisation and neuroprotection. Free Radic Biol Med, 75(Suppl 1): S16, 2014.
34.
KrauseM, BockPM, TakahashiHK, Homem De BittencourtPIJr., and NewsholmeP. The regulatory roles of NADPH oxidase, intra- and extra-cellular HSP70 in pancreatic islet function, dysfunction and diabetes. Clin Sci (Lond), 128: 789–803, 2015.
35.
LambethJD and NeishAS. Nox enzymes and new thinking on reactive oxygen: a double-edged sword revisited. Annu Rev Pathol, 9: 119–145, 2014.
LiJ, WangJJ, and ZhangSX. NADPH oxidase 4-derived H2O2 promotes aberrant retinal neovascularization via activation of VEGF receptor 2 pathway in oxygen-induced retinopathy. J Diabetes Res, 2015: 963289, 2015.
38.
LiN, LiB, BrunT, Deffert-DelbouilleC, MahioutZ, DaaliY, MaXJ, KrauseKH, and MaechlerP. NADPH oxidase NOX2 defines a new antagonistic role for reactive oxygen species and cAMP/PKA in the regulation of insulin secretion. Diabetes, 61: 2842–2850, 2012.
39.
MacDonaldMJ, AmmonHP, PatelT, and SteinkeJ. Failure of 6-aminonicotinamide to inhibit the potentiating effect of leucine and arginine on glucose-induced insulin release in vitrol. Diabetologia, 10: 761–765, 1974.
40.
MacDonaldPE, JosephJW, and RorsmanP. Glucose-sensing mechanisms in pancreatic beta-cells. Philos Trans R Soc Lond B Biol Sci, 360: 2211–2225, 2005.
41.
Milo-LandesmanD, SuranaM, BerkovichI, CompagniA, ChristoforiG, FleischerN, and EfratS. Correction of hyperglycemia in diabetic mice transplanted with reversibly immortalized pancreatic beta cells controlled by the tet-on regulatory system. Cell Transplant, 10: 645–650, 2001.
42.
MohammedAM and KowluruA. Activation of apocynin-sensitive NADPH oxidase (Nox2) activity in INS-1 832/13 cells under glucotoxic conditions. Islets, 5: 129–131, 2013.
43.
MorganD, Oliveira-EmilioHR, KeaneD, HirataAE, Santos da RochaM, BordinS, CuriR, NewsholmeP, and CarpinelliAR. Glucose, palmitate and pro-inflammatory cytokines modulate production and activity of a phagocyte-like NADPH oxidase in rat pancreatic islets and a clonal beta cell line. Diabetologia, 50: 359–369, 2007.
44.
MorganD, RebelatoE, AbdulkaderF, GracianoMF, Oliveira-EmilioHR, HirataAE, RochaMS, BordinS, CuriR, and CarpinelliAR. Association of NAD(P)H oxidase with glucose-induced insulin secretion by pancreatic beta-cells. Endocrinology, 150: 2197–2201, 2009.
45.
NewsholmeP, MorganD, RebelatoE, Oliveira-EmilioHC, ProcopioJ, CuriR, and CarpinelliA. Insights into the critical role of NADPH oxidase(s) in the normal and dysregulated pancreatic beta cell. Diabetologia, 52: 2489–2498, 2009.
46.
PandeyD and FultonDJ. Molecular regulation of NADPH oxidase 5 via the MAPK pathway. Am J Physiol Heart Circ Physiol, 300: H1336–H1344, 2011.
47.
PandeyD, GrattonJP, RafikovR, BlackSM, and FultonDJ. Calcium/calmodulin-dependent kinase II mediates the phosphorylation and activation of NADPH oxidase 5. Mol Pharmacol, 80: 407–415, 2011.
48.
PandeyD, PatelA, PatelV, ChenF, QianJ, WangY, BarmanSA, VenemaRC, SteppDW, RudicRD, and FultonDJ. Expression and functional significance of NADPH oxidase 5 (Nox5) and its splice variants in human blood vessels. Am J Physiol Heart Circ Physiol, 302: H1919–H1928, 2012.
49.
PapaFR. Endoplasmic reticulum stress, pancreatic beta-cell degeneration, and diabetes. Cold Spring Harb Perspect Med, 2: a007666, 2012.
50.
PiJ, BaiY, ZhangQ, WongV, FloeringLM, DanielK, ReeceJM, DeeneyJT, AndersenME, CorkeyBE, and CollinsS. Reactive oxygen species as a signal in glucose-stimulated insulin secretion. Diabetes, 56: 1783–1791, 2007.
51.
PrentkiM, VischerS, GlennonMC, RegazziR, DeeneyJT, and CorkeyBE. Malonyl-CoA and long chain acyl-CoA esters as metabolic coupling factors in nutrient-induced insulin secretion. J Biol Chem, 267: 5802–5810, 1992.
52.
PyneNJ and FurmanBL. Cyclic nucleotide phosphodiesterases in pancreatic islets. Diabetologia, 46: 1179–1189, 2003.
53.
RebelatoE, Mares-GuiaTR, GracianoMF, LabriolaL, BrittoLR, Garay-MalpartidaHM, CuriR, SogayarMC, and CarpinelliAR. Expression of NADPH oxidase in human pancreatic islets. Life Sci, 91: 244–249, 2012.
54.
RohE, SongDK, and KimMS. Emerging role of the brain in the homeostatic regulation of energy and glucose metabolism. Exp Mol Med, 48: e216, 2016.
55.
RotherE, BelgardtBF, TsaousidouE, HampelB, WaismanA, MyersMGJr., and BruningJC. Acute selective ablation of rat insulin promoter-expressing (RIPHER) neurons defines their orexigenic nature. Proc Natl Acad Sci U S A, 109: 18132–18137, 2012.
56.
SchmidtHH, StockerR, VollbrachtC, PaulsenG, RileyD, DaiberA, and CuadradoA. Antioxidants in translational medicine. Antioxid Redox Signal, 23: 1130–1143, 2015.
57.
SchrenzelJ, SerranderL, BanfiB, NusseO, FouyouziR, LewDP, DemaurexN, and KrauseKH. Electron currents generated by the human phagocyte NADPH oxidase. Nature, 392: 734–737, 1998.
58.
SzkudelskiT and SzkudelskaK. Regulatory role of adenosine in insulin secretion from pancreatic beta-cells—action via adenosine A(1) receptor and beyond. J Physiol Biochem, 71: 133–140, 2015.
59.
Taylor-FishwickDA. NOX, NOX who is there?. The contribution of NADPH oxidase one to beta cell dysfunction. Front Endocrinol (Lausanne), 4: 40, 2013.
60.
TomasA, YermenB, MinL, PessinJE, and HalbanPA. Regulation of pancreatic beta-cell insulin secretion by actin cytoskeleton remodelling: role of gelsolin and cooperation with the MAPK signalling pathway. J Cell Sci, 119: 2156–2167, 2006.
61.
TouyzRM, AnagnostopoulouA, CamargoLL, RiosFJ, and MontezanoAC. Vascular biology of superoxide-generating NADPH oxidase 5-implications in hypertension and cardiovascular disease. Antioxid Redox Signal, 30: 1027–1040, 2019.
WangL, OplandD, TsaiS, LukCT, SchroerSA, AllisonMB, EliaAJ, FurlongerC, SuzukiA, PaigeCJ, MakTW, WinerDA, MyersMGJr., and WooM. Pten deletion in RIP-Cre neurons protects against type 2 diabetes by activating the anti-inflammatory reflex. Nat Med, 20: 484–492, 2014.
64.
WeaverJR, HolmanTR, ImaiY, JadhavA, KenyonV, MaloneyDJ, NadlerJL, RaiG, SimeonovA, and Taylor-FishwickDA. Integration of pro-inflammatory cytokines, 12-lipoxygenase and NOX-1 in pancreatic islet beta cell dysfunction. Mol Cell Endocrinol, 358: 88–95, 2012.
65.
WojtusciszynA, AndresA, MorelP, CharvierS, ArmanetM, TosoC, ChoiY, BoscoD, and BerneyT. Immunomodulation by blockade of the TRANCE co-stimulatory pathway in murine allogeneic islet transplantation. Transpl Int, 22: 931–939, 2009.
66.
ZywertA, SzkudelskaK, and SzkudelskiT. Effects of adenosine A(1) receptor antagonism on insulin secretion from rat pancreatic islets. Physiol Res, 60: 905–911, 2011.
Supplementary Material
Please find the following supplemental material available below.
For Open Access articles published under a Creative Commons License, all supplemental material carries the same license as the article it is associated with.
For non-Open Access articles published, all supplemental material carries a non-exclusive license, and permission requests for re-use of supplemental material or any part of supplemental material shall be sent directly to the copyright owner as specified in the copyright notice associated with the article.
