Proteomics characterization of cell model with renal fibrosis phenotype: osmotic stress as fibrosis triggering factor.
Renal fibroblasts are thought to play a major role in the development of renal fibrosis (RF). The mechanisms leading to this renal alteration remain poorly understood. We performed differential proteomic analyses with two established fibroblast cell lines with RF phenotype to identify new molecular pathways associated with RF. Differential 2-DE combined with mass spectrometry analysis revealed the alteration of more than 30 proteins in fibrotic kidney fibroblasts (TK188) compared to normal kidney fibroblast (TK173). Among these proteins, markers of the endoplasmic reticulum (ER) stress- and the unfolded protein response (UPR) pathway (GRP78, GRP94, ERP57, ERP72, and CALR) and the oxidative stress pathway proteins (PRDX1, PRDX2, PRDX6, HSP70, HYOU1) were highly up-regulated in fibrotic cells. Activation of these stress pathways through long time exposition of TK173, to high NaCl or glucose concentrations resulted in TK188 like phenotype. Parallel to an increase in reactive oxygen species, the stressed cells showed significant alteration of fibrosis markers, ER-stress and oxidative stress proteins. Similar effects of osmotic stress could be also observed on renal proximal tubule cells. Our data suggest an important role of the ER-stress proteins in fibrosis and highlights the pro-fibrotic effect of osmotic stress through activation of oxidative stress and ER-stress pathways.
Department of Nephrology and Rheumatology, Georg-August University Goettingen, Goettingen, Germany. dihazi@med.uni-goettingen.de
Hassan
H
Dihazi
Gry Helene
GH
Dihazi
Claudia
C
Mueller
Loubna
L
Lahrichi
Abdul R
AR
Asif
Asima
A
Bibi
Marwa
M
Eltoweissy
Radovan
R
Vasko
Gerhard A
GA
Mueller
D002460:Cell Line; D015180:Electrophoresis, Gel, Two-Dimensional; D004721:Endoplasmic Reticulum / Q000378:metabolism; D000091342:Endoplasmic Reticulum Chaperone BiP; D005347:Fibroblasts / Q000378:metabolism; D005355:Fibrosis / Q000473:pathology*; D019084:Fluorescent Antibody Technique, Indirect; D006801:Humans; D007668:Kidney / Q000378:metabolism / Q000473:pathology*; D009995:Osmosis; D018384:Oxidative Stress; D010641:Phenotype; D040901:Proteomics / Q000379:methods*; D017382:Reactive Oxygen Species; D015854:Up-Regulation
D000091342:Endoplasmic Reticulum Chaperone BiP; C000717247:HSPA5 protein, human; D017382:Reactive Oxygen Species