POL Scientific / Bladder / Volume 11 / Issue 2 / DOI: 10.14440/bladder.2024.0011
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RESEARCH ARTICLE

Effect of pro-inflammatory cytokines on urothelial cell adenosine triphosphate release and breakdown

Belinda Kaleska1 ,  Ronald Sluyter2 ,  Zhuoran Chen3 ,  Kylie J Mansfield1*
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1 Graduate School of Medicine, Faculty of Science, Medicine and Health, University of Wollongong, Wollongong, NSW 2522, Australia
2 Molecular Horizons and School of Chemistry and Molecular Biosciences, University of Wollongong, Wollongong, NSW 2522, Australia
3 Department of Urogynaecology, St George Hospital, University of New South Wales, Kogarah, NSW 2522, Australia
Bladder 2024 , 11(2), e21200006; https://doi.org/10.14440/bladder.2024.0011
Submitted: 30 June 2024 | Accepted: 20 August 2024 | Published: 12 September 2024
© 2024 by the Bladder published by POL Scientific. Licensee POL Scientific, USA. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution 4.0 International License ( https://creativecommons.org/licenses/by/4.0/ )
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Abstract

Objectives: Urinary symptoms of urgency, frequency, and pain are thought to be the result of inflammation in several bladder pathologies although the cause of these symptoms remains uncertain. Extracellular adenosine triphosphate (ATP) released from the bladder urothelium during normal bladder stretch is believed to bind to purinergic receptors on afferent nerves to signal bladder sensation. This study examined pro-inflammatory cytokines in the urine of women with detrusor overactivity (DO) with or without urinary tract infection (UTI) compared to controls and then determined the effect of pro-inflammatory cytokines on ATP signaling (release and breakdown) from the urothelium. Methods: The urinary concentrations of interferon-gamma (IFN-γ), tumor necrosis factor-alpha (TNF-α), and interleukin-1 beta (IL-1β) were determined in women with DO with or without UTI compared to female controls. The effect of pro-inflammatory cytokines (IFN-γ, TNF-α, and IL-1β) on control and hypotonic-induced ATP release using human UROtsa urothelial cells was examined, as was the effect of these cytokines on nucleotide (ATP, adenosine diphosphate and adenosine monophosphate) breakdown. Results: Urinary concentrations of IFN-γ, TNF-α, and IL-1β were increased in women with DO and UTI. Pre-treatment of urothelial cells with individual cytokines stimulated a decrease rather than an increase in ATP release whereas pre-treatment with a cocktail of all three cytokines stimulated a small but significant increase in hypotonic-induced ATP release. Pre-treatment of urothelial cells with cytokines significantly enhanced nucleotide breakdown. Conclusion: Using a simple cell culture model we have demonstrated that the response of the urothelium to pro-inflammatory cytokines is complex, affecting both release and breakdown of ATP.

Keywords
Urothelium
Extracellular adenosine triphosphate
Inflammation
Cytokine
Ectonucleotides
Funding
The cytokine assays for this project were funded by the Illawarra Health and Medical Research Institute (IHMRI).
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Conflict of interest
The authors declare no conflict of interest.
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