Abstract: TH-PO856
Inhibition of NLRP3 Inflammasome Blocks Peritoneal Dialysis Solution-Induced Mesothelial-to-Mesenchymal Transition
Session Information
- Peritoneal Dialysis - I
November 02, 2017 | Location: Hall H, Morial Convention Center
Abstract Time: 10:00 AM - 10:00 AM
Category: Dialysis
- 608 Peritoneal Dialysis
Authors
- Zhang, Lu, the affiliated hospital of Nanjing University of Chinese Medicine, Nanjing, Jiangsu, China
- Zheng, Min, the affiliated hospital of Nanjing University of Chinese Medicine, Nanjing, Jiangsu, China
- Zhou, Dong, University of Pittsburgh, Pittsburgh, Pennsylvania, United States
- Gao, Kun, the affiliated hospital of Nanjing University of Chinese Medicine, Nanjing, Jiangsu, China
- He, Weiming, the affiliated hospital of Nanjing University of Chinese Medicine, Nanjing, Jiangsu, China
- Li, Qing, the affiliated hospital of Nanjing University of Chinese Medicine, Nanjing, Jiangsu, China
- Li, Zhenghong, the affiliated hospital of Nanjing University of Chinese Medicine, Nanjing, Jiangsu, China
- Sheng, Meixiao, the affiliated hospital of Nanjing University of Chinese Medicine, Nanjing, Jiangsu, China
- Sun, Wei, the affiliated hospital of Nanjing University of Chinese Medicine, Nanjing, Jiangsu, China
Background
To investigate the role of activated Nod-like Receptor Protein 3 (NLRP3) inflammasome in peritoneal dialysis (PD) solution-induced mesothelial-mesenchymal transition (MMT) in peritoneal mesothelial cells (PMC).
Methods
1.Human peritoneal mesothelial cells (HMrSV5) were incubated with different concentrations of PD solution (1.5%，2.5%，4.25%) for 24h or 4.25% PD solution for various periods of time at 0, 6, 12, 24, 48 hours, respectively. The supernatant was collected for ELISA assay and cells were lysed using RIPA buffer for a western blot assay.2. After stable transfection of siRNA plasmid targeting NLRP3, HMrSV5 were incubated with 4.25% PD solution for 24h, cells were then harvested for western blot assay.3. HMrSV5 were treated with 4.25% PD solution for 24h in the absence or presence of different doses of Astragaloside IV (As-IV),which is a cycloartane triterpene saponin with a clear formula(C41H68O14).
Results
1. The PD solution promotes IL-18 secretion in the condition medium of cultured PMCs as dose- and time-dependent manner. Accordingly, it significantly induced NLRP3, pro-IL-1β, IL-1β, pro-casepase-1, and caspase-1 expression in vitro. 2. The PD solution induces MMT which is featured with decreased E-cadherin and increased vimentin, α-SMA. 3. Knockdown NLRP3 partially preserve PMCs from MMT by inhibited IL-18 secretion and NLRP3, caspase-1, pro-IL-1β, IL-1β expression in cultured PMC. 4. Therapeutic inhibition of NLRP3 inflammasome with a novel small molecule inhibitor, As-IV, preserves mesothelial phenotypes in the established model in vitro.
Conclusion
The activated NLRP3 inflammasome mediates PD solution-induced MMT in PMCs. Targeting NLRP3 inflammasome activity by small molecule inhibitor AS-IV abolishes inflammatory factors and blocks mesenchymal conversion of mesothelium.
Funding
- Government Support - Non-U.S.