A Proteomic View on the Role of Glucose in Peritoneal Dialysis

Abstract

Peritoneal dialysis is a frequently used mode of renal replacement therapy although peritoneal dialysis fluid (PDF) acts as stressor for mesothelial cells. In this study, stress response to PDF is investigated by a proteomics approach using Met-5A cell cultures closely resembling mesothelial cells. In a previous work, we identified about 100 proteins as significantly enhanced or diminished in abundance after full-PDF stress (90 mM glucose, pH 5.8, and presence of lactate and glucose degradation products (GDPs)) using two-dimensional electrophoresis (2-DE) and MALDI-MS and MS/MS techniques. In this paper, a functional analysis is presented assigning these proteins to glucose associated pathways according to the KEGG database. To establish the stressor role of high glucose concentration, the up/down regulation of proteins populating these pathways were investigated in a fluorescence-difference gel electrophoresis (DIGE) experiment exposing Met-5A cells to nonphysiologically high glucose conditions only. In this glucose-single stress experiment, the fold-change ratios of the investigated glucose-pathway associated proteins were found much lower than observed in the previous full-PDF stress experiments. This finding supports the hypothesis that cellular response to full-PDF stress is not primarily induced by the high glucose concentration, even when focusing on proteins belonging to the glucose associated pathways.

Transcriptional control of glyoxalase 1 by Nrf2 provides a stress-responsive defence against dicarbonyl glycation

“…It has an established role in the enzymatic defence against protein and nucleotide glycation by dicarbonyls [28] but transcriptional control of Glo1 by a stress responsive mechanism has not hitherto been disclosed. Induction of Glo1 expression has been found in cells exposed to metabolic and inflammatory stress in vitro and in vivo: glomerular cells challenged by increased MG formation in hyperglycaemia associated with diabetes [29], mesothelial cells challenged by MG and high glucose concentrations of clinical dialysis fluids [30], and inflammatory stress induced by amyloidosis of Alzheimer's disease [31] and overexpression of α-synuclein in an experimental model of Parkinson's disease [32]. In these cases of increased cell dysfunction and damage and physiological morbidity, Glo1 induction was insufficient to prevent increased protein damage by MG.…”

Characterization and proteomic profile of extracellular vesicles from peritoneal dialysis efflux

“…Several studies have searched for biomarkers in PDE correlating with PM function, detection of fibrosis, and/or the failure of the technique (reviewed in[25]). Proteomic studies of mesothelial cell lines[7] and transcriptome analysis in rats[26] have reported differences between the protein and miRNA content, respectively, of cells exposed or non-exposed to peritoneal liquids.…”

Proteomics in Peritoneal Dialysis

“…Thus, peritoneal dialysis effluent is considered as a significant stressor for the MC layer (Breborowicz et al 1995;Wieslander et al 1991). To address this issue, a cell line derived from the MC layers was used as a model to study the glucoserelated pathways induced by high concentration of glucose (Lechner et al 2010). Using twodimensional fluorescence-difference gel electrophoresis (DIGE), altered proteins upon glucose stress in Met-5A cell were revealed.…”

“…Protein isoforms are distinguished by numbers in brackets. [Adapted from (Lechner et al 2010)] Another laboratory works on analyzing the protein composition of peritoneal fluid from patients receiving peritoneal dialysis with different concentration of glucose (Cuccurullo et al 2010). Peritoneal dialysis effluent with different glucose concentrations were revealed by 2DE.…”