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.