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Energy depletion and not ROS formation is a crucial step of glucolipotoxicity (GLTx) in pancreatic beta cells

  • Molecular and cellular mechanisms of disease
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Abstract

We have shown previously that genetic or pharmacological deletion of KATP channels protect against beta cell dysfunction induced by reactive oxygen species (ROS). Since it is assumed that glucolipotoxicity (GLTx) causes ROS production, we aimed to evaluate whether suppression of KATP channel activity can also prevent beta cell damage evoked by GLTx. We used an in vitro model of GLTx and measured distinct parameters of stimulus-secretion coupling. GLTx gradually induced disturbances of Ca2+ oscillations over 3 days. This impairment in Ca2+ dynamics was partially reversed in beta cells without functional KATP channels (SUR1−/−) and by the sulfonylurea gliclazide but not by tolbutamide. By contrast, the GLTx-induced suppression of glucose-induced insulin secretion could not be rescued by decreased KATP channel activity pointing to a direct interaction of GLTx with the secretory capacity. Accordingly, GLTx also suppressed KCl-induced insulin secretion. GLTx was not accompanied by decisively increased ROS production or enhanced apoptosis. Insulin content of beta cells was markedly reduced by GLTx, an effect not prevented by gliclazide. Since GLTx markedly diminished the mitochondrial membrane potential and cellular ATP content, lack of ATP is assumed to decrease insulin biosynthesis. The deleterious effect of GLTx is therefore caused by direct interference with the secretory capacity whereby reduction of insulin content is one important parameter. These findings deepen our understanding how GLTx damages beta cells and reveal that GLTx is disconnected from ROS formation, a notion important for targeting beta cells in the treatment of diabetes. Overall, GLTx-induced energy depletion may be a primary step in the cascade of events leading to loss of beta cell function in type-2 diabetes mellitus.

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Abbreviations

[Ca2+]c :

Cytosolic calcium concentration

2-OH-E+ :

2-Hydroxyethidium

DCF:

2′,7′-Dichlorofluorescein

DIV:

Days in vitro

GLTx:

Glucolipotoxicity

ROS:

Reactive oxygen species

SSC:

Stimulus-secretion coupling

T2DM:

Type 2 diabetes mellitus

ΔΨ:

Mitochondrial membrane potential

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Acknowledgements

This work was supported by grants from the Deutsche Forschungsgemeinschaft (GD; Dr225/11-1) and the Brazilian National Council of Technological and Scientific Development (CNPq) (MBO; scholarship number 490162/2013-4).

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Authors and Affiliations

  1. Institute of Pharmacy, Department of Pharmacology, University of Tübingen, Auf der Morgenstelle 8, 72076, Tübingen, Germany

    Morgana Barroso Oquendo, Nikolas Layer, Rebecca Wagner, Peter Krippeit-Drews & Gisela Drews

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  1. Morgana Barroso Oquendo
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  2. Nikolas Layer
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Correspondence to Gisela Drews.

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Barroso Oquendo, M., Layer, N., Wagner, R. et al. Energy depletion and not ROS formation is a crucial step of glucolipotoxicity (GLTx) in pancreatic beta cells. Pflugers Arch - Eur J Physiol 470, 537–547 (2018). https://doi.org/10.1007/s00424-017-2094-8

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