PD14-11 NOX-DERIVED SUPEROXIDE PRODUCTION IN BLADDER UROTHELIUM – EFFECT OF NOX SUBTYPE DELETION AND THE INFLAMMATORY FACTORS

You have accessJournal of UrologyBladder & Urethra: Anatomy, Physiology & Pharmacology II (PD14)1 Apr 2019PD14-11 NOX-DERIVED SUPEROXIDE PRODUCTION IN BLADDER UROTHELIUM – EFFECT OF NOX SUBTYPE DELETION AND THE INFLAMMATORY FACTORS Maxwell W Roberts, Josephine Amosah, Guiping Sui, Rui Wu, Simon Archer, Michael Ruggieri, and Changhao Wu* Maxwell W RobertsMaxwell W Roberts More articles by this author , Josephine AmosahJosephine Amosah More articles by this author , Guiping SuiGuiping Sui More articles by this author , Rui WuRui Wu More articles by this author , Simon ArcherSimon Archer More articles by this author , Michael RuggieriMichael Ruggieri More articles by this author , and Changhao Wu*Changhao Wu* More articles by this author View All Author Informationhttps://doi.org/10.1097/01.JU.0000555468.89302.e9AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVES: Recognition of the urothelium as a new sensory structure and its potential role in mediating bladder dysfunction has led to great interest in identifying its pathological mediators. Oxidative stress is a fundamental pathological mediator and ROS-generating enzyme NADPH-oxidase (Nox) has particular importance as it is the only enzyme in the body that produces ROS as its sole function and can serve as specific ROS-controlling target without compromising normal biochemical oxidation. Our recent study has identified such system in bladder urothelium (1). To explore molecular mechanisms of superoxide production and the pathological significance of Nox-associated oxidative stress, we examined the effect of Nox-subtype deletion and several pathologically important inflammatory factors on Nox-derived superoxide production in bladder urothelial tissue. METHODS: Wild type C57BL/6J mice, Nox-1 and Nox-2 knockout mice and their wild type littermates were used in compliance with the regulations. Bladder mucosa and full-thickness bladder tissue were isolated under microscopic guidance. NADPH-dependent superoxide production in live tissue was determined by lucigenin-enhanced chemiluminescence. RESULTS: Nox-1 deletion reduced the superoxide production to 64.7±7.5% of wild type littermate control (mean±SEM; n=6, p<0.01), showing Nox-1 contribution to endogenous superoxide production. Nox-2 deletion reduced the superoxide level to 78.0± 5.8 % of the control (n=9, p<0.01), demonstrating Nox-2 contribution. Angiotensin II (1µM), a trophic factor and vessel constrictor which is endogenously released and involved in bladder outflow obstruction and hypertrophy, increased the superoxide production from 85.6±6.3 to 138±11.7 RLU/mg tissue (n=16, p<0.01). TRPV4-specific activator GSK1016790A (1µM), which participates in sensory dysfunction and bladder pain, enhanced the superoxide production in mucosa-attached bladder strips from 89.7±5.5 to 118.8 ±5.6 RLU/mg tissue (n=10, p<0.01). Endothelin-1 (1µM), an inflammatory factor endogenously released in tissue ischemia, also augmented the superoxide production in bladder mucosa from 44.5±7.6 to 90.2±17.9 RLU/mg tissue (n=10, p<0.05). CONCLUSIONS: These new data show that Nox-1 and Nox-2 subtypes contribute to endogenous superoxide production in bladder urothelium. The results also provide the first evidence that inflammatory factors angiotensin II, GSK1016790A and endothelin-1 can upregulate the Nox-derived superoxide production in bladder mucosa and demonstrate the pathological significance of Nox-associated oxidative stress. Source of Funding: BBSRC BB/P004695/1; NIA 1R01AG049321-01A1 Reference Roberts M, Amosah J, Sui G, Wu R, Archer S, Ruggieri, M and Wu C (2018). Identifying the functional subtypes of NADPH oxidases (Nox) from the urothelium: implication for ROS-controlling targets. J Urol 99, S, e1061. Guildford, United Kingdom; London, United Kingdom; Coventry, United Kingdom; Guildford, United Kingdom; Philadelphia, PA; Guildford, United Kingdom© 2019 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 201Issue Supplement 4April 2019Page: e235-e235 Advertisement Copyright & Permissions© 2019 by American Urological Association Education and Research, Inc.MetricsAuthor Information Maxwell W Roberts More articles by this author Josephine Amosah More articles by this author Guiping Sui More articles by this author Rui Wu More articles by this author Simon Archer More articles by this author Michael Ruggieri More articles by this author Changhao Wu* More articles by this author Expand All Advertisement PDF downloadLoading ...