Abstract: SA-PO729
The Influence of Alanyl-Glutamine on the Peritoneal Proteome in a Chronic Model of Peritoneal Dialysis
Session Information
- Peritoneal Dialysis - II
November 04, 2017 | Location: Hall H, Morial Convention Center
Abstract Time: 10:00 AM - 10:00 AM
Category: Dialysis
- 608 Peritoneal Dialysis
Authors
- Boehm, Michael, Medical University Vienna, Vienna, Austria
- Herzog, Rebecca, Medical University Vienna, Vienna, Austria
- Aufricht, Christoph, Medical University Vienna, Vienna, Austria
- Kratochwill, Klaus, Medical University Vienna, Vienna, Austria
Background
Peritoneal dialysis (PD) fluids exert cytotoxic properties towards peritoneal mesothelial cells. Recent studies showed that alanyl-glutamine (Ala-Gln) modulates the cellular stress response, improves survival of mesothelial cells and reduced submesothelial thickening in experimental models of PD, and improved PD effluent cell function with regards to stress and immune responses in clinical studies. However, the mechanism of Ala-Gln-mediated membrane protection is not yet fully understood. Here, we apply a novel proteomics approach in a clinically relevant in vivo model.
Methods
Following experimental PD for 5 weeks, using PD fluid with or without supplementation of Ala-Gln, or sham treatment as control, mesothelial cells from rat peritoneum were directly harvested by detergent extraction and subjected to proteomic analysis based on difference gel electrophoresis (DiGE) and subsequent protein identification by mass spectrometry. Submesothelial thickening was measured in parallel by microscopy of peritoneal tissue sections.
Results
From a total protein spot pattern of 744 spots, in 500 spots proteins were identified, linking to 233 unique protein IDs. Using database information from UniProt, proteins were assigned either to the group of high abundance plasma proteins (serum-type) or to the group of cellular proteins (cell-type). Following statistical analysis of mixed effects of PD fluid and Ala-Gln, pathway analysis allowed further assignment of candidate proteins to specific roles in the stress response and membrane preservation, observed by decreased submesothelial thickening.
Conclusion
This study shows that by combination of proteomics and bioinformatics the separation of residual plasma proteins and cellular proteins of various origin in PD effluent is feasible. Thereby, the peritoneal mesothelial cell stress proteome can be assessed after exposure to different PD fluids. Identified biological processes might allow linking molecular mechanisms of membrane protection in the in vivo model to protective effects observed in vitro and in clinical PD.