Differential Expression of Proteins From Cultured Endothelial Cells Exposed to Uremic Versus Normal Serum
Received 24 May 2007; accepted 28 November 2007.
Background
Deficient hemostasis and accelerated atherosclerosis coexist in patients with chronic kidney disease. Endothelial dysfunction may be involved in the high incidence of atherothrombotic events in these patients. We established an in vitro model of endothelial dysfunction by exposing endothelial cells to uremic media and applied a proteomic approach to characterize endothelial cell dysfunction in uremia.
Study Design
Cross-sectional study.
Setting and Participants
Serum samples from 8 patients with chronic kidney disease on hemodialysis treatment were collected.
Predictor
Exposure of cultured endothelial cells to normal and uremic serum.
Outcome and Measurements
Proteins from lysed cells were characterized by isoelectric point and molecular weight by using 2-dimensional gel electrophoresis. Spots were visualized by means of silver staining and identified by using mass spectrometry.
Results
Identification of the most prominent proteins showed molecules related to inflammation (high mobility group box 1, aldose reductase, and proteasome components) and oxidative stress (superoxide dismutase and glutathione peroxidase), both associated with chronic kidney disease. These changes may be caused by activation of the nuclear factor-κB transcription factor. Changes in expression of cytoskeletal proteins (destrin and vimentin) also were detected.
Limitations
In vitro study.
Conclusion
Proteomic techniques proved to be a powerful tool to investigate endothelial dysfunction in uremia. A more exhaustive analysis will provide answers and potential therapeutic targets in the near future.
1Servicio de Hemoterapia-Hemostasia y, Centre de Diagnòstic Biomèdic, Institut d’Investigacions biomèdiques August Pi i Sunyer (IDIBAPS), Universitat de Barcelona, Barcelona, Spain
2Servicio de Nefrología, Centre de Diagnòstic Biomèdic, Institut d’Investigacions biomèdiques August Pi i Sunyer (IDIBAPS), Universitat de Barcelona, Barcelona, Spain
3Centro Superior de Investigaciones Científicas–Universidad Autónoma de Barcelona (CSIC-UAB) Proteomics Facility, Instituto de Investigaciones Biomédicas de Barcelona-CSIC/IDIBAPS, Barcelona, Spain.
Address correspondence to Maribel Díaz-Ricart, PhD, Servicio de Hemoterapia-Hemostasia, Hospital Clínic, Villarroel 170, 08036 Barcelona, Spain.
ABSTRACT