Proteomics analysis identifies PARK7 as an important player for renal cell resistance and survival under oxidative stress
Proteomics analysis identifies PARK7 as an important player for renal cell resistance and survival under oxidative stress
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DOI：
10.1039/c0mb00116c
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发表时间：
2011-01-01
影响因子：
--
通讯作者：
Dihazi, Hassan
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文献类型：
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作者：
Eltoweissy, Marwa;Mueller, Gerhard A.;Dihazi, Hassan
肾纤维化是一个以排泄肾功能下降为特征的过程。纤维化的分子机制尚未完全了解。据报道，氧化应激途径参与肾组织恶化和纤维化进展。为了鉴定与氧化应激和肾纤维化相关的新的分子靶点，用已建立的肾细胞系（TK 173和HK-2）进行差异蛋白质组学分析。用氧化应激触发因子H（2）O（2）处理细胞，并研究蛋白质组的变化。二维蛋白质图谱的产生和差异表达的蛋白质进行处理和鉴定，使用质谱分析结合数据库搜索。有趣的是，在H（2）O（2）处理后，肾细胞系中ROS的增加伴随着大量蛋白质的改变，这些蛋白质可以分为三类：第一类是已被描述为参与纤维形成的蛋白质（例如ACTA 2、VIN、Vim、DES、KRT、COL 1A 1、COL 4A 1），第二类更有趣，涉及氧化应激途径的蛋白质（PRDX 1，PRDX 2，PRDX 6，SOD，PARK 7，HYOU 1），它们在氧化应激下高度上调，第三类代表蛋白质，它们参与不同的其他代谢途径。在氧化应激蛋白中，PARK 7的上调伴随着氧化导致的pI的变化。使用siRNA敲低PARK 7导致在氧化应激下肾细胞活力的显著降低。在H2 O2处理下，PARK 7基因敲减的细胞与对照组相比，细胞活力下降了80%，细胞凋亡增加。这些结果首次强调了PARK 7在肾细胞抗氧化应激中的重要作用。
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.