重组人粒细胞集落刺激因子对脑缺血再灌注大鼠大脑皮质蛋白质的影响

目的：研究造血干细胞动员剂重组人粒细胞集落刺激因子（rhG-CSF）对脑缺血神经系统具有保护作用的蛋白质组学机制。方法：本实验利用脑缺血再灌注大鼠模型（tMCAO模型）在再灌注2h后颈部皮下注射rhG-CSF，在灌注后14d提取大脑皮质蛋白进行双向电泳。结果：缺血再灌注损伤（模型组）大鼠与假手术组大鼠比较，筛选到56个差异表达蛋白质点，其中17个上调，39个下调，应用基质辅助激光解吸电离飞行时间质谱仪（MALDI-TOF-MS）得到肽质量指纹图，输入美国国立生物技术信息中心（NCBI）蛋白质数据库进行检索，并在Swiss-Prot数据库中进一步验证，鉴定出其中19个为已知蛋白质。而经过人粒细胞集落刺激因子（G-CSF）治疗（G-CSF治疗组）大鼠与模型组大鼠比较，筛选出35个蛋白质点，其中16个下调，19个上调，其中鉴定为已知蛋白质的有6个，分别为二氢嘧啶酶相关蛋白2、胶质纤维酸性蛋白、内皮黏蛋白、RhoGDP解离抑制因子、RabGDP解离抑制因子、鸟嘌呤核苷酸结合蛋白。结论：脑缺血后大鼠脑组织蛋白质表达发生变化，G-CSF在脑缺血亚急性期可能通过调节多种神经再生相关蛋白质的表达，参与保护脑缺血的神经元。

Effect of recombined granulocyte colony-stimulating factor on cerebral cortical proteins of ischemia-reperfusion injury rats

Objective： To research the relation between difference protein with neuron protective effect of G-CSF in ischemia-reperfusion rats brain by using proteomics.Methods： Twenty-four experimental Wistar rats were divide into 3 groups, control group, rats cured by G-CSF group （G-CSF group）, ischemia-reperfusion injured rats group （I/R group）. Ischemia-reperfusion injury rats model was generated by using Koizumi’s way in G group and I/R group, one nylon thread was used to block the rats middle cerebral artery and reperfused after 2 hours. G-CSF （50μg/kg/d） was injected subcutaneously on rats’ backs for successive 5 days in G-CSF group. Rats were executed and decapitated after 14 days after reperfusion to get the brain respectively. Sodium chloride injection was used in I/R group and control group.Cortex proteins were extracted in the 3 groups of rats. Then the maps of the proteins were established by DIGE （differential gel electrophoresis, DIGE）. The altered protein spots were identiifed with MALDI-TOF-MS and database searching.Results： Compared with the contrl group, the I/R group gained 56 differential protein spots, 39 spots expressed lowly, and 17 spots high. Compared with I/R group, the G-SCF group gained 35 differential protein spots, 16 spots expressed lowly, and 19 spots high, identiifed 6 protein spots, including dihydropyrimidinase-associated protein 2, glial ifbrillary acidic protein, endo-mucin, Rho GDP dissociation inhibitor, Rab GDP dissociation inhibitor and guanine-nucleotide-binding protein.＆amp;nbsp;Conclusion： G-CSF is involved in neuroprotection after brain ischemia, possibly by regulating the expression of various neural regeneration-associated proteins at the subacute stage.