Proteomics analysis identifies PARK7 as an important player for renal cell resistance and survival under oxidative stress.

Marwa Eltoweissy,Claudia A. Müller,Asima Bibi,Phuc Van Nguye,Gry H. Dihazi,Gerhard A. Müller,Hassan Dihazi

doi:10.1039/c0mb00116c

Marwa Eltoweissy, Claudia A. Müller + Show 5 more

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https://doi.org/10.1039/c0mb00116c

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Abstract

Renal fibrosis is a process that is characterized by declining excretory renal function. The molecular mechanisms of fibrosis are not fully understood. Oxidative stress pathways were reported to be involved in renal tissue deterioration and fibrosis progression. In order to identify new molecular targets associated with oxidative stress and renal fibrosis, differential proteomics analysis was performed with established renal cell lines (TK173 and HK-2). The cells were treated with oxidative stress triggering factor H(2)O(2) and the proteome alterations were investigated. Two dimensional protein maps were generated and differentially expressed proteins were processed and identified using mass spectrometry analysis combined with data base search. Interestingly the increase of ROS in the renal cell lines upon H(2)O(2) treatment was accompanied by alteration of a large number of proteins, which could be classified in three categories: the first category grouped the proteins that have been described to be involved in fibrogenesis (e.g. ACTA2, VIN, VIM, DES, KRT, COL1A1, COL4A1), the second category, which was more interesting involved proteins of the oxidative stress pathway (PRDX1, PRDX2, PRDX6, SOD, PARK7, HYOU1), which were highly up-regulated under oxidative stress, and the third category represented proteins, which are involved in different other metabolic pathways. Among the oxidative stress proteins the up-regulation of PARK7 was accompanied by a shift in the pI as a result of oxidation. Knockdown of PARK7 using siRNA led to significant reduction in renal cell viability under oxidative stress. Under H(2)O(2) treatment the PARK7 knockdown cells showed up to 80% decrease in cell viability and an increase in apoptosis compared to the controls. These results highlight for the first time the important role of PARK7 in oxidative stress resistance in renal cells.

Highlights

reactive oxygen species’’ (ROS) production beyond the organ’s scavenging capacity simultaneously attacks and alters other target molecules and causes lipid peroxidation induction, DNA breakdown and/or proteins denaturation.[2]A growing body of evidence suggests oxidative stress as one of the most relevant pathogenic influences in numerous human diseases, including diabetes, vascular complications,[3] cancer,[4] and neurodegenerative disorders such as Alzheimer’s5 and Parkinson’s disease.[6]
We attempted to assess the response of renal fibroblast cells stimulated with H2O2 over 24 h using the MTT cell viability assay
To detect and identify altered protein expression induced by H2O2 exposure, we employed conventional 2-DE coupled with mass spectrometry analysis

Summary

Introduction

A growing body of evidence suggests oxidative stress as one of the most relevant pathogenic influences in numerous human diseases, including diabetes, vascular complications,[3] cancer,[4] and neurodegenerative disorders such as Alzheimer’s5 and Parkinson’s disease.[6] Oxidative stress has been reported in association with the occurrence of impaired reproductive function,[7] and as a major cause of liver damage,[8] and cardiac myocyte death.[9] Oxidative stress plays a fundamental role in cataractogenesis,[10] it contributes to mucosal inflammation of the gastrointestinal tract[11] and is important in aging processes.[12] There is convincing experimental and clinical evidence that ROS generation is an important fibrogenic. The molecular pathogenesis of renal interstitial fibrosis correlates with the degree of renal functional loss and predicts the rate of progression to end-stage renal failure.[13]

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