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Calcium Pyrophosphate And Monosodium Urate Activate The NLRP3 Inflammasome Within Bladder Urothelium Via Reactive Oxygen Species And TXNIP.

dc.contributor.author Harper, Shelby N
dc.contributor.author Leidig, Patrick D
dc.contributor.author Hughes, Francis M
dc.contributor.author Jin, Huixia
dc.contributor.author Purves, J Todd
dc.date.accessioned 2020-08-06T22:22:19Z
dc.date.available 2020-08-06T22:22:19Z
dc.date.issued 2019-01
dc.identifier 225767
dc.identifier.issn 2253-2447
dc.identifier.issn 2253-2447
dc.identifier.uri https://hdl.handle.net/10161/21292
dc.description.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.
dc.language eng
dc.publisher Informa UK Limited
dc.relation.ispartof Research and reports in urology
dc.relation.isversionof 10.2147/RRU.S225767
dc.subject NLRP3 inflammasome
dc.subject ROS
dc.subject TXNIP
dc.subject inflammation
dc.subject stones
dc.subject urothelium
dc.title Calcium Pyrophosphate And Monosodium Urate Activate The NLRP3 Inflammasome Within Bladder Urothelium Via Reactive Oxygen Species And TXNIP.
dc.type Journal article
duke.contributor.id Leidig, Patrick D|0656505
duke.contributor.id Hughes, Francis M|0686657
duke.contributor.id Purves, J Todd|0685081
dc.date.updated 2020-08-06T22:22:17Z
pubs.begin-page 319
pubs.end-page 325
pubs.organisational-group School of Medicine
pubs.organisational-group Surgery, Urology
pubs.organisational-group Duke
pubs.organisational-group Surgery
pubs.organisational-group Clinical Science Departments
pubs.organisational-group Staff
pubs.publication-status Published
pubs.volume 11
duke.contributor.orcid Hughes, Francis M|0000-0003-3776-3653
duke.contributor.orcid Purves, J Todd|0000-0001-9689-2047


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