Indoleamine 2,3-dioxygenase controls purinergic receptor-mediated ischemia-reperfusion injury in renal tubular epithelial cells

Theodoros Eleftheriadis; Georgios Pissas; Spyridon Golfinopoulos; Vassilios Liakopoulos; Ioannis Stefanidis

doi:10.1515/jbcpp-2022-0128

Article

Indoleamine 2,3-dioxygenase controls purinergic receptor-mediated ischemia-reperfusion injury in renal tubular epithelial cells

Theodoros Eleftheriadis , Georgios Pissas , Spyridon Golfinopoulos , Vassilios Liakopoulos and Ioannis Stefanidis

Published/Copyright: August 2, 2022

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From the journal Journal of Basic and Clinical Physiology and Pharmacology Volume 34 Issue 6

Abstract

Objectives

Ischemia–reperfusion (I–R) injury is the most common cause of acute kidney injury (AKI). Experimental studies have shown that indoleamine 2,3-dioxygenase 1 (IDO) and the purinergic receptor P2X7 contribute to kidney I–R injury. We evaluated whether there is an interplay between IDO and P2X7 in the pathogenesis of I–R injury.

Methods

Primary renal proximal tubular epithelial cells (RPTECs) were subjected to anoxia or reoxygenation with or without specific inhibitors. Cell imaging, colorimetric assays, and Western blotting were used.

Results

Cell imaging revealed that inhibition of IDO, or all the purinergic receptors with an ATPase, or specific inhibition of P2X7 rescued the cells from anoxia or reoxygenation-induced cell death. This was confirmed with LDH release assay, which also detected the ferroptotic nature of cell death due to reoxygenation. On the contrary, activated cleaved caspase 3 increased during anoxia, showing that apoptosis prevails. All the aforementioned treatments prevented caspase increase. Both anoxia and reoxygenation increased extracellular ATP, IDO, and P2X7 expression. IDO remained unaffected by the above-mentioned treatments. On the contrary, treatment with apyrase or inhibition of P2X7decreased extracellular ATP and P2X7 expression, which are also decreased by inhibition of IDO. The first indicates a positive feedback loop regarding P2X7 activation, expression and function, while the latter implies that IDO controls P2X7 expression.

Conclusions

In RPRECs subjected to anoxia or reoxygenation, IDO is upregulated, increasing P2X7 and contributing to anoxia or reoxygenation-induced cell death. Clarifying the molecular mechanisms implicated in kidney I–R injury is of particular interest since it may lead to new therapeutic strategies against AKI.

Keywords: acute kidney injury; indoleamine 2,3-dioxygenase; ischemia-reperfusion injury; P2X7

Corresponding author: Theodoros Eleftheriadis, MD, PhD, Department of Nephrology, Faculty of Medicine, University of Thessaly, Biopolis, Mezourlo Hill, 41110 Larissa, Greece, Phone: 00302413501665, Fax: 00302413501667, E-mail: teleftheriadis@yahoo.com

Theodoros Eleftheriadis and Georgios Pissas contributed equally.

Research funding: None, the project was supported by the resources of our department.
Author contribution: T.E. designed the study, T.E. and G.P. performed the experiments, T.E., G.P., S.G., V.L., and I.S. analyzed the results, T.E. wrote the manuscript with help from G.P. I.S. supported all stages.
Competing interests: The authors declare that they have no competing interests.
Availability of data and materials: The analyzed datasets generated during the study are available from the corresponding author on reasonable request.
Research involving Human Participants and/or Animals: Not applicable.

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Supplementary Material

The online version of this article offers supplementary material (https://doi.org/10.1515/JBCPP-2022-0128).

Received: 2022-05-09

Accepted: 2022-07-03

Published Online: 2022-08-02

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https://doi.org/10.1515/jbcpp-2022-0128

Keywords for this article

acute kidney injury; indoleamine 2,3-dioxygenase; ischemia-reperfusion injury; P2X7