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Inhibition of NADPH oxidase-1 preserves beta cell function

Abstract

Aims/hypothesis

Upregulation of the reactive oxygen species (ROS)-producing enzyme NADPH oxidase (NOX)-1 in islets and beta cells follows acute exposure to inflammatory cytokines and is concomitant with beta cell dysfunction. NOX-1 is a candidate mediator of inflammation-induced beta cell dysfunction. This study aimed to determine whether selective inhibition of NADPH oxidase-1 presents a new strategy to preserve beta cell function.

Methods

Induced beta cell dysfunction was studied in primary human donor islets, isolated mouse islets and murine beta cell lines. Islets and beta cells were stimulated with inflammatory cytokines (TNF-α, IL-1β, IFN-γ). NOX-1 activity was blocked by the selective inhibitor ML171.

Results

Cytokine induction of intracellular ROS was reduced 80% with 1 μmol/l ML171 in murine beta cell lines (p < 0.01). Cytokine-induced apoptosis, measured by caspase-3 activation or quantified fluorescence microscopy, was prevented in islets and beta cell lines up to 100% with ML171 in a concentration-dependent manner (p < 0.05). Functionally, glucose-stimulated insulin secretion was abolished by cytokine exposure but preserved by ML171 in isolated mouse islets and murine beta cell lines. A feed-forward regulation of NOX-1 in islets and beta cell lines was disrupted by ML171.

Conclusions/interpretation

Stimulation of NOX-1 activity is a major component of inflammatory cytokine-induced beta cell dysfunction. Significant protection of beta cells is conferred with selective inhibition of NOX-1. Suppression of NOX-1 activity may present a new therapeutic strategy to preserve and protect beta cell function in diabetes.

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Abbreviations

C57BL/6J:

C57 black 6

DCF-DA:

6-Carboxy-2′,7′-dichlorodihydrofluorescein diacetate, di(acetoxymethyl ester)

GADPH:

Glyceraldehyde 3-phosphate dehydrogenase

GSIS:

Glucose-stimulated insulin secretion

NOX:

NADPH oxidase

PIC:

Proinflammatory cytokine

RFU:

Relative fluorescence unit

ROS:

Reactive oxygen species

TBS:

Tris-buffered saline

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Acknowledgements

Human donor islets were provided by the Integrated Islet Distribution Program (IIDP, http://iipd.coh.org/).

Funding

Funding for this work was provided by CDMRP, Department of Defense (PR093521, DAT-F), JDRF International and the Leona M. and Harry B. Helmsley Charitable Trust (DAT-F).

Duality of interest

The authors declare that there is no duality of interest associated with this manuscript.

Contribution statement

DAT-F directed the work, provided data interpretation and wrote the article. JRW and WJG acquired research data, contributed to the analysis and interpretation of data and helped to draft and/or review the article. All authors approved this manuscript version. DAT-F is responsible for the integrity of the work as a whole.

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Correspondence to David A. Taylor-Fishwick.

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Weaver, J.R., Grzesik, W. & Taylor-Fishwick, D.A. Inhibition of NADPH oxidase-1 preserves beta cell function. Diabetologia 58, 113–121 (2015). https://doi.org/10.1007/s00125-014-3398-2

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  • DOI: https://doi.org/10.1007/s00125-014-3398-2

Keywords

  • Apoptosis
  • Beta cells
  • Cytokines
  • Diabetes
  • Inflammation
  • ML171
  • NADPH oxidase-1