Department of Neurology, the Second Affiliated Hospital of Dalian Medical University, Dalian, Liaoning 116023, P.R. China.
Int J Mol Med. 2013 Sep;32(3):607-14. doi: 10.3892/ijmm.2013.1419. Epub 2013 Jun 18.
Endothelial progenitor cells (EPCs) repair vascular damage and participate in neovascularization. Accumulating evidence has demonstrated that EPCs have therapeutic potential in reactive oxygen species (ROS)-mediated vascular diseases. In this study, to investigate the effects of oxidative stress on EPCs, EPCs were treated with H2O2 at different final concentrations for 3 h. MTT assay, scratch-wound assay and Matrigel invasion assay revealed that cell proliferation, migration and tubule formation and function, respectively, were impaired under H2O2 stress in a concentration-dependent manner. To determine protein response to H2O2 stress, two-dimensional differential in-gel electrophoresis (2D-DIGE) combined with matrix-assisted laser desorption-ionization time-of-flight (MALDI-TOF/TOF) mass spectrometry were performed. The results revealed that triosephosphate isomerase and ADP-sugar pyrophosphatase were downregulated, while peroxiredoxin-2, thioredoxin-dependent peroxide reductase, mitochondrial (Prx‑3), peroxiredoxin-6, EGF-containing fibulin-like extracellular matrix protein 1, vimentin and Rab GDP dissociation inhibitor α were upregulated in the H2O2-treated EPCs. To further confirm the results from mass spectrometry, the expression pattern of Prx-3 in response to H2O2 stress was examined by western blot analysis. The data presented in this study provide novel insight into the defensive mechanisms of EPCs and the pathways of oxidative damage in an oxidative environment.
内皮祖细胞 (EPCs) 可修复血管损伤并参与血管新生。越来越多的证据表明，EPCs 在活性氧 (ROS) 介导的血管疾病中具有治疗潜力。在这项研究中，为了研究氧化应激对 EPCs 的影响，用不同终浓度的 H2O2 处理 EPCs 3 小时。MTT 检测、划痕实验和基质胶侵袭实验表明，EPCs 在 H2O2 应激下，细胞增殖、迁移和小管形成及功能分别呈浓度依赖性受损。为了确定蛋白质对 H2O2 应激的反应，采用二维差异凝胶电泳 (2D-DIGE) 结合基质辅助激光解吸电离飞行时间 (MALDI-TOF/TOF) 质谱进行分析。结果表明，磷酸丙糖异构酶和 ADP-糖磷酸水解酶下调，而过氧化物酶-2、硫氧还蛋白依赖的过氧化物还原酶、线粒体 (Prx-3)、过氧化物酶-6、含 EGF 的纤维连接蛋白样细胞外基质蛋白 1、波形蛋白和 Rab GDP 解离抑制剂α在 H2O2 处理的 EPCs 中上调。为了进一步证实质谱的结果，通过 Western blot 分析检测了 Prx-3 对 H2O2 应激的表达模式。本研究提供了内皮祖细胞防御机制和氧化环境中氧化损伤途径的新见解。
