Calcium Pyrophosphate And Monosodium Urate Activate The NLRP3 Inflammasome Within Bladder Urothelium Via Reactive Oxygen Species And TXNIP.
Abstract
Objective:To investigate the in vitro activation of the NLRP3 inflammasome within
bladder urothelium by stone-forming components. Further, to describe the contributions
of reactive oxygen species (ROS) and thioredoxin-interacting protein (TXNIP), an important
structural component of the inflammasome, to this activation. Methods:Urothelial cells
were harvested and incubated overnight. For agonist studies, cells were treated with
varying concentrations of calcium pyrophosphate (CPPD) and monosodium urate (MSU).
For inhibitor studies, cells were treated with either N-acetylcysteine (NAC) (1 hr)
or Verapamil (4 hrs) prior to incubation with either CPPD (62.5 ug/mL) or MSU (1.25
ug/mL) for 24 hrs. Untreated controls were incubated with ATP (1.25 mM) for 1 hr to
maximally stimulate NLRP3 inflammasome activity (measured as caspase-1 cleavage of
the fluorogenic substrate Ac-YVAD-AFC). Results are reported as a percentage of maximum
ATP response. Results:CPPD and MSU activate caspase-1 in urothelial cells in a dose-dependent
manner, reaching ~50% and ~25% of the ATP response, respectively. Pre-treatment with
the general ROS scavenger NAC reduces this activation in a dose-dependent manner.
Additionally, activation was suppressed through treatment with Verapamil, a known
downregulator of TXNIP expression. Conclusion:The stone components CPPD and MSU activate
NLRP3 in an ROS and TXNIP-dependent manner in bladder urothelium. These findings demonstrate
the importance of ROS and TXNIP, and suggest that targeting either may be a way to
decrease stone-dependent NLRP3 inflammation within the bladder.
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https://hdl.handle.net/10161/21292Published Version (Please cite this version)
10.2147/RRU.S225767Publication Info
Harper, Shelby N; Leidig, Patrick D; Hughes, Francis M; Jin, Huixia; & Purves, J Todd (2019). Calcium Pyrophosphate And Monosodium Urate Activate The NLRP3 Inflammasome Within
Bladder Urothelium Via Reactive Oxygen Species And TXNIP. Research and reports in urology, 11. pp. 319-325. 10.2147/RRU.S225767. Retrieved from https://hdl.handle.net/10161/21292.This is constructed from limited available data and may be imprecise. To cite this
article, please review & use the official citation provided by the journal.
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Show full item recordScholars@Duke
Monty Hughes Jr.
Assistant Professor in Urology
Dr. Hughes received his Ph.D. from the Medical University of South Carolina and was
a post doc at both the University of North Carolina at Chapel Hill and NIH. He then
joined the faculty of the University of North Carolina at Charlotte where he rose
to the rank of Associate Professor (with tenure). Following a brief stint as the director
of the biology division of a start-up pharmaceutical company, he joined forces with
Dr. Purves at the Medical University of South Carolina to begin this l
Patrick Leidig
House Staff
J Todd Purves
Professor of Urology
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