Related Experiment Video
Updated: Aug 13, 2025
Real-Time Void Spot Assay
Published on: February 10, 2023
Selective Pharmacological Inhibition of NOX2 by GSK2795039 Improves Bladder Dysfunction in Cyclophosphamide-Induced
Mariana G de Oliveira1, Fabíola Z Monica1, Gabriela R Passos1
1Department of Translational Medicine, Pharmacology Area, Faculty of Medical Sciences, University of Campinas (UNICAMP), Alexander Fleming St., Campinas 13083-881, SP, Brazil.
Abstract:
Interstitial cystitis/bladder pain syndrome (IC/BPS) is a chronic inflammatory disease without consistently effective treatment. Among the many mediators implicated in cystitis, the overproduction of reactive oxygen species (ROS) seems to play a key role, although the main source of ROS remains unclear. This study aimed to investigate the contribution of NADPH oxidase (NOX) isoforms in ROS generation and the voiding dysfunction of cyclophosphamide (CYP, 300 mg/Kg, ip, 24 h)-induced cystitis in adult female mice, a well-recognized animal model to study IC/BPS, by using GKT137831 (5 mg/Kg, ip, three times in a 24 h period) or GSK2795039 (5 mg/Kg, ip, three times in a 24 h period) to inhibit NOX1/4 or NOX2, respectively. Our results showed that treatment with GSK2795039 improved the dysfunctional voiding behavior induced by CYP, reduced bladder edema and inflammation, and preserved the urothelial barrier integrity and tight junction occludin expression, besides inhibiting the characteristic vesical pain and bladder superoxide anion generation. In contrast, the NOX1/4 inhibitor GKT137831 had no significant protective effects. Taken together, our in vivo and ex vivo data demonstrate that NOX2 is possibly the main source of ROS observed in cystitis-induced CYP in mice. Therefore, selective inhibition of NOX2 by GSK2795039 may be a promising target for future therapies for IC/BPS.
Insights
Reactive oxygen species (ROS) contribute to interstitial cystitis/bladder pain syndrome (IC/BPS). Targeting NADPH oxidase 2 (NOX2) with GSK2795039 effectively reduced pain and inflammation in a mouse model.
Area of Science:
- Urology
- Inflammation Research
- Pharmacology
Background:
- Interstitial cystitis/bladder pain syndrome (IC/BPS) is a chronic condition with limited treatment options.
- Reactive oxygen species (ROS) are implicated in IC/BPS pathogenesis, but their primary source is unknown.
- NADPH oxidase (NOX) enzymes are potential sources of ROS in inflammatory conditions.
Purpose of the Study:
- To investigate the role of NADPH oxidase (NOX) isoforms in generating reactive oxygen species (ROS).
- To evaluate the therapeutic potential of NOX inhibitors in a cyclophosphamide (CYP)-induced mouse model of IC/BPS.
- To identify the specific NOX isoform responsible for ROS production in cystitis.
Main Methods:
- Induction of cystitis in female mice using cyclophosphamide (CYP).
- Administration of NOX1/4 inhibitor (GKT137831) or NOX2 inhibitor (GSK2795039).
- Assessment of voiding behavior, bladder inflammation, urothelial integrity, and ROS generation.
Main Results:
- GSK2795039 treatment significantly improved voiding function, reduced bladder edema and inflammation, and preserved urothelial barrier integrity.
- GSK2795039 inhibited superoxide anion generation in the bladder and reduced vesical pain.
- GKT137831 (NOX1/4 inhibitor) showed no significant protective effects.
Conclusions:
- NADPH oxidase 2 (NOX2) is identified as a major source of ROS in CYP-induced cystitis.
- Selective inhibition of NOX2 by GSK2795039 demonstrates therapeutic promise for IC/BPS.
- Targeting NOX2 offers a potential new strategy for managing IC/BPS symptoms.
