Brea David, Rodríguez-González Raquel, Sobrino Tomás, Rodríguez-Yañez Manuel, Blanco Miguel, Castillo José
Hospital Clínico Universitario, IDIS, University of Santiago de Compostela, Spain.
Neurol Res. 2011 Dec;33(10):1057-63. doi: 10.1179/1743132811Y.0000000038.
An increase in the circulating concentration of endothelial progenitor cells (EPCs) is associated with a better outcome in patients with acute ischemic stroke. Likewise, EPCs are heterogeneous cells, with functional differences and different protein expressions. Our objective was to compare protein expressions of EPCs from ischemic stroke patients and healthy subjects.
Eleven ischemic stroke patients and 11 healthy subjects, matched by age and gender, were included in this study. EPC colonies were defined as early outgrowth colony forming unit-endothelial cell. Cells were lysed and proteins were purified and separated on two-dimensional gels. Gel images were analyzed using the PDQuest software and protein differences between EPCs from ischemic stroke patients and healthy subjects were identified by mass spectrometry. Results were finally validated by western blot.
Proteomic analysis revealed three qualitative differences between EPCs from healthy subjects and ischemic stroke patients. Two of them, endoplasmatic reticulum protein-29 and CdC-42, were only expressed in EPCs from healthy subjects, whereas elongation factor-2 was only identified in EPCs from ischemic stroke patients. Furthermore, we identified one protein, peroxiredoxin-1, whose expression was 10 times stronger in ischemic stroke patients than in healthy subjects. Western blot analysis showed greater expression of endoplasmatic reticulum protein-29 in EPCs from healthy subjects and elongation factor-2 and peroxiredoxin-1 in EPCs from ischemic stroke patients.
Proteomic analysis showed differences in protein expressions of EPCs from ischemic stroke patients and healthy subjects that may be involved in mechanisms related to functional impairment.
循环内皮祖细胞(EPCs)浓度升高与急性缺血性中风患者较好的预后相关。同样,EPCs是异质性细胞,具有功能差异和不同的蛋白表达。我们的目的是比较缺血性中风患者和健康受试者EPCs的蛋白表达。
本研究纳入了11例缺血性中风患者和11例年龄及性别相匹配的健康受试者。EPC集落被定义为早期生长集落形成单位-内皮细胞。细胞裂解后,蛋白被纯化并在二维凝胶上分离。使用PDQuest软件分析凝胶图像,并通过质谱鉴定缺血性中风患者和健康受试者EPCs之间的蛋白差异。结果最终通过蛋白质印迹法进行验证。
蛋白质组学分析揭示了健康受试者和缺血性中风患者EPCs之间的三个定性差异。其中两个,内质网蛋白-29和Cdc-42,仅在健康受试者的EPCs中表达,而延伸因子-2仅在缺血性中风患者的EPCs中鉴定到。此外,我们鉴定出一种蛋白,过氧化物酶1,其在缺血性中风患者中的表达比健康受试者强10倍。蛋白质印迹分析显示内质网蛋白-29在健康受试者的EPCs中表达更高,而延伸因子-2和过氧化物酶1在缺血性中风患者的EPCs中表达更高。
蛋白质组学分析显示缺血性中风患者和健康受试者EPCs的蛋白表达存在差异,这些差异可能与功能损害相关的机制有关。