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Diabetologia
Published in: Diabetologia 5/2013

Open Access 01-05-2013 | Article

NADPH oxidase inhibition prevents beta cell dysfunction induced by prolonged elevation of oleate in rodents

Authors: K. Koulajian, T. Desai, G. C. Liu, A. Ivovic, J. N. Patterson, C. Tang, J. El-Benna, J. W. Joseph, J. W. Scholey, A. Giacca

Published in: Diabetologia | Issue 5/2013

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Abstract

Aims/hypothesis

The activation of NADPH oxidase has been implicated in NEFA-induced beta cell dysfunction. However, the causal role of this activation in vivo remains unclear. Here, using rodents, we investigated whether pharmacological or genetic inhibition of NADPH oxidase could prevent NEFA-induced beta cell dysfunction in vivo.

Methods

Normal rats were infused for 48 h with saline or oleate with or without the NADPH oxidase inhibitor apocynin. In addition, NADPH oxidase subunit p47phox-null mice and wild-type littermate controls were infused with saline or oleate for 48 h. This was followed by measurement of NADPH oxidase activity, reactive oxygen species (ROS) and superoxide imaging and assessment of beta cell function in isolated islets and hyperglycaemic clamps.

Results

Oleate infusion in rats increased NADPH oxidase activity, consistent with increased total but not mitochondrial superoxide in islets and impaired beta cell function in isolated islets and during hyperglycaemic clamps. Co-infusion of apocynin with oleate normalised NADPH oxidase activity and total superoxide levels and prevented beta cell dysfunction. Similarly, 48 h NEFA elevation in wild-type mice increased total but not mitochondrial superoxide and impaired beta cell function in isolated islets. p47phox-null mice were protected against these effects when subjected to 48 h oleate infusion. Finally, oleate increased the levels of total ROS, in both models, whereas inhibition of NADPH oxidase prevented this increase, suggesting that NADPH oxidase is the main source of ROS in this model.

Conclusions/interpretation

These data show that NADPH-oxidase-derived cytosolic superoxide is increased in islets upon oleate infusion in vivo; and whole-body NADPH-oxidase inhibition decreases superoxide in concert with restoration of islet function.
Appendix
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Literature
1.
Giacca A, Xiao C, Oprescu AI, Carpentier AC, Lewis GF (2011) Lipid-induced pancreatic beta-cell dysfunction: focus on in vivo studies. Am J Physiol Endocrinol Metab 300:E255–E262PubMedCrossRef
2.
Moore PC, Ugas MA, Hagman DK, Parazzoli SD, Poitout V (2004) Evidence against the involvement of oxidative stress in fatty acid inhibition of insulin secretion. Diabetes 53:2610–2616PubMedCrossRef
3.
Morgan D, Oliveira-Emilio HR, Keane D et al (2007) 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 50:359–369PubMedCrossRef
4.
Syed I, Jayaram B, Subasinghe W, Kowluru A (2010) Tiam1/Rac1 signaling pathway mediates palmitate-induced, ceramide-sensitive generation of superoxides and lipid peroxides and the loss of mitochondrial membrane potential in pancreatic beta-cells. Biochem Pharmacol 80:874–883PubMedCrossRef
5.
Carlsson C, Borg LA, Welsh N (1999) Sodium palmitate induces partial mitochondrial uncoupling and reactive oxygen species in rat pancreatic islets in vitro. Endocrinology 140:3422–3428PubMedCrossRef
6.
Maestre I, Jordan J, Calvo S et al (2003) Mitochondrial dysfunction is involved in apoptosis induced by serum withdrawal and fatty acids in the beta-cell line INS-1. Endocrinology 144:335–345PubMedCrossRef
7.
Wang X, Li H, de Leo D et al (2004) Gene and protein kinase expression profiling of reactive oxygen species-associated lipotoxicity in the pancreatic beta-cell line MIN6. Diabetes 53:129–140PubMedCrossRef
8.
Lenzen S, Drinkgern J, Tiedge M (1996) Low antioxidant enzyme gene expression in pancreatic islets compared with various other mouse tissues. Free Radic Biol Med 20:463–466PubMedCrossRef
9.
Maechler P, Jornot L, Wollheim CB (1999) Hydrogen peroxide alters mitochondrial activation and insulin secretion in pancreatic beta cells. J Biol Chem 274:27905–27913PubMedCrossRef
10.
Oprescu AI, Bikopoulos G, Naassan A et al (2007) Free fatty acid-induced reduction in glucose-stimulated insulin secretion: evidence for a role of oxidative stress in vitro and in vivo. Diabetes 56:2927–2937PubMedCrossRef
11.
Zhang X, Bao Y, Ke L, Yu Y (2010) Elevated circulating free fatty acids levels causing pancreatic islet cell dysfunction through oxidative stress. J Endocrinol Investig 33:388–394
12.
Xiao C, Giacca A, Lewis GF (2008) Oral taurine but not N-acetylcysteine ameliorates NEFA-induced impairment in insulin sensitivity and beta cell function in obese and overweight, non-diabetic men. Diabetologia 51:139–146PubMedCrossRef
13.
Oliveira HR, Verlengia R, Carvalho CR, Britto LR, Curi R, Carpinelli AR (2003) Pancreatic beta-cells express phagocyte-like NAD(P)H oxidase. Diabetes 52:1457–1463PubMedCrossRef
14.
Uchizono Y, Takeya R, Iwase M et al (2006) Expression of isoforms of NADPH oxidase components in rat pancreatic islets. Life Sci 80:133–139PubMedCrossRef
15.
Nakayama M, Inoguchi T, Sonta T et al (2005) Increased expression of NAD(P)H oxidase in islets of animal models of Type 2 diabetes and its improvement by an AT1 receptor antagonist. Biochem Biophys Res Commun 332:927–933PubMedCrossRef
16.
Babior BM, Lambeth JD, Nauseef W (2002) The neutrophil NADPH oxidase. Arch Biochem Biophys 397:342–344PubMedCrossRef
17.
Mason TM, Goh T, Tchipashvili V et al (1999) Prolonged elevation of plasma free fatty acids desensitizes the insulin secretory response to glucose in vivo in rats. Diabetes 48:524–530PubMedCrossRef
18.
Liu GC, Fang F, Zhou J et al (2012) Deletion of p47phox attenuates the progression of diabetic nephropathy and reduces the severity of diabetes in the Akita mouse. Diabetologia 55:2522–2532PubMedCrossRef
19.
Kimura S, Zhang GX, Nishiyama A et al (2005) Role of NAD(P)H oxidase- and mitochondria-derived reactive oxygen species in cardioprotection of ischemic reperfusion injury by angiotensin II. Hypertension 45:860–866PubMedCrossRef
20.
Diao J, Allister EM, Koshkin V et al (2008) UCP2 is highly expressed in pancreatic alpha-cells and influences secretion and survival. Proc Natl Acad Sci USA 105:12057–12062PubMedCrossRef
21.
Tang C, Han P, Oprescu AI et al (2007) Evidence for a role of superoxide generation in glucose-induced beta-cell dysfunction in vivo. Diabetes 56:2722–2731PubMedCrossRef
22.
LeBel CP, Ischiropoulos H, Bondy SC (1992) Evaluation of the probe 2′,7′-dichlorofluorescin as an indicator of reactive oxygen species formation and oxidative stress. Chem Res Toxicol 5:227–231PubMedCrossRef
23.
Bilski P, Belanger AG, Chignell CF (2002) Photosensitized oxidation of 2′,7′-dichlorofluorescin: singlet oxygen does not contribute to the formation of fluorescent oxidation product 2′,7′-dichlorofluorescein. Free Radic Biol Med 33:938–946PubMedCrossRef
24.
DeFronzo RA, Tobin JD, Andres R (1979) Glucose clamp technique: a method for quantifying insulin secretion and resistance. Am J Physiol 237:E214–E223PubMed
25.
Goldstein BJ, Mahadev K, Wu X, Zhu L, Motoshima H (2005) Role of insulin-induced reactive oxygen species in the insulin signaling pathway. Antioxid Redox Signal 7:1021–1031PubMedCrossRef
26.
Morgan D, Rebelato E, Abdulkader F et al (2009) Association of NAD(P)H oxidase with glucose-induced insulin secretion by pancreatic beta-cells. Endocrinology 150:2197–2201PubMedCrossRef
27.
Newsholme P, Morgan D, Rebelato E et al (2009) Insights into the critical role of NADPH oxidase(s) in the normal and dysregulated pancreatic beta cell. Diabetologia 52:2489–2498PubMedCrossRef
28.
Pi J, Bai Y, Zhang Q et al (2007) Reactive oxygen species as a signal in glucose-stimulated insulin secretion. Diabetes 56:1783–1791PubMedCrossRef
29.
Yuan L, Li X, Xu GL, Qi CJ (2010) Effects of renin-angiotensin system blockade on islet function in diabetic rats. J Endocrinol Investig 33:13–19
30.
Zhu CF, Peng HB, Liu GQ, Zhang F, Li Y (2010) Beneficial effects of oligopeptides from marine salmon skin in a rat model of type 2 diabetes. Nutrition 26:1014–1020PubMedCrossRef
31.
Harmon JS, Stein R, Robertson RP (2005) Oxidative stress-mediated, post-translational loss of MafA protein as a contributing mechanism to loss of insulin gene expression in glucotoxic beta cells. J Biol Chem 280:11107–11113PubMedCrossRef
32.
Kaneto H, Xu G, Fujii N, Kim S, Bonner-Weir S, Weir GC (2002) Involvement of c-Jun N-terminal kinase in oxidative stress-mediated suppression of insulin gene expression. J Biol Chem 277:30010–30018PubMedCrossRef
33.
Michalska M, Wolf G, Walther R, Newsholme P (2010) Effects of pharmacological inhibition of NADPH oxidase or iNOS on pro-inflammatory cytokine, palmitic acid or H2O2-induced mouse islet or clonal pancreatic beta-cell dysfunction. Biosci Rep 30:445–453PubMedCrossRef
34.
Newsholme P, Haber EP, Hirabara SM et al (2007) Diabetes associated cell stress and dysfunction: role of mitochondrial and non-mitochondrial ROS production and activity. J Physiol 583:9–24PubMedCrossRef
35.
Santos LR, Rebelato E, Graciano MF, Abdulkader F, Curi R, Carpinelli AR (2011) Oleic acid modulates metabolic substrate channeling during glucose-stimulated insulin secretion via NAD(P)H oxidase. Endocrinology 152:3614–3621PubMedCrossRef
36.
Valle MM, Graciano MF, Lopes de Oliveira ER et al (2011) Alterations of NADPH oxidase activity in rat pancreatic islets induced by a high-fat diet. Pancreas 40:390–395PubMedCrossRef
37.
Meng R, Zhu DL, Bi Y, Yang DH, Wang YP (2010) Apocynin improves insulin resistance through suppressing inflammation in high-fat diet-induced obese mice. Mediat Inflamm 2010:858735CrossRef
38.
Meng R, Zhu DL, Bi Y, Yang DH, Wang YP (2011) Anti-oxidative effect of apocynin on insulin resistance in high-fat diet mice. Ann Clin Lab Sci 41:236–243PubMed
39.
Yuzefovych L, Wilson G, Rachek L (2010) Different effects of oleate vs. palmitate on mitochondrial function, apoptosis, and insulin signaling in L6 skeletal muscle cells: role of oxidative stress. Am J Physiol Endocrinol Metab 299:E1096–E1105PubMedCrossRef
40.
Pike LS, Smift AL, Croteau NJ, Ferrick DA, Wu M (2011) Inhibition of fatty acid oxidation by etomoxir impairs NADPH production and increases reactive oxygen species resulting in ATP depletion and cell death in human glioblastoma cells. Biochim Biophys Acta 1807:726–734PubMedCrossRef
41.
Iizuka K, Nakajima H, Namba M et al (2002) Metabolic consequence of long-term exposure of pancreatic beta cells to free fatty acid with special reference to glucose insensitivity. Biochim Biophys Acta 1586:23–31PubMedCrossRef
42.
Schonfeld P, Wojtczak L (2008) Fatty acids as modulators of the cellular production of reactive oxygen species. Free Radic Biol Med 45:231–241PubMedCrossRef
43.
Ivarsson R, Quintens R, Dejonghe S et al (2005) Redox control of exocytosis: regulatory role of NADPH, thioredoxin, and glutaredoxin. Diabetes 54:2132–2142PubMedCrossRef
44.
MacDonald PE, Salapatek AM, Wheeler MB (2003) Temperature and redox state dependence of native Kv2.1 currents in rat pancreatic beta-cells. J Physiol 546:647–653PubMedCrossRef
45.
Subasinghe W, Syed I, Kowluru A (2011) Phagocyte-like NADPH oxidase promotes cytokine-induced mitochondrial dysfunction in pancreatic beta-cells: evidence for regulation by Rac1. Am J Physiol Regul Integr Comp Physiol 300:R12–R20PubMedCrossRef
46.
Heumuller S, Wind S, Barbosa-Sicard E et al (2008) Apocynin is not an inhibitor of vascular NADPH oxidases but an antioxidant. Hypertension 51:211–217PubMedCrossRef
47.
Stolk J, Hiltermann TJ, Dijkman JH, Verhoeven AJ (1994) Characteristics of the inhibition of NADPH oxidase activation in neutrophils by apocynin, a methoxy-substituted catechol. Am J Respir Cell Mol Biol 11:95–102PubMedCrossRef
48.
Fontayne A, Dang PM, Gougerot-Pocidalo MA, El-Benna J (2002) Phosphorylation of p47phox sites by PKC alpha, beta II, delta, and zeta: effect on binding to p22phox and on NADPH oxidase activation. Biochemistry 41:7743–7750PubMedCrossRef
49.
Miwa I, Ichimura N, Sugiura M, Hamada Y, Taniguchi S (2000) Inhibition of glucose-induced insulin secretion by 4-hydroxy-2-nonenal and other lipid peroxidation products. Endocrinology 141:2767–2772PubMedCrossRef
Metadata
Title
NADPH oxidase inhibition prevents beta cell dysfunction induced by prolonged elevation of oleate in rodents
Authors
K. Koulajian
T. Desai
G. C. Liu
A. Ivovic
J. N. Patterson
C. Tang
J. El-Benna
J. W. Joseph
J. W. Scholey
A. Giacca
Publication date
01-05-2013
Publisher
Springer-Verlag
Published in
Diabetologia / Issue 5/2013
Print ISSN: 0012-186X
Electronic ISSN: 1432-0428
DOI
https://doi.org/10.1007/s00125-013-2858-4

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