期刊
RESEARCH AND REPORTS IN UROLOGY
卷 11, 期 -, 页码 319-325出版社
DOVE MEDICAL PRESS LTD
DOI: 10.2147/RRU.S225767
关键词
stones; urothelium; inflammation; NLRP3 inflammasome; TXNIP; ROS
资金
- American Urological Association
- Duke University School of Medicine
- National Institute of Diabetes and Digestive and Kidney Diseases of the National Institutes of Health, USA [R01DK103534, R01DK117890]
- Duke University Medical Center, Department of Surgery, Division of Urology
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 similar to 50% and similar to 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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