卒中后G-CSF对骨髓干细胞的动员和神经修复作用.docVIP

卒中后G-CSF对骨髓干细胞的动员和神经修复作用 张兴秀1 郭慧娟1 李琳2 王健1 郑敏＃2 （1重庆医科大学附属第二医院，2重庆医科大学基础医学院 #通讯作者） 摘 要 目的 研究粒细胞集落刺激因子(G-CSF)对卒中后骨髓干细胞的动员与血管再生和神经修复的影响。 方法 线栓法制备大鼠大脑中动脉栓塞(MCAO)模型，成功造模大鼠分为生理盐水对照组和G-CSF治疗组。治疗组皮下注射G-CSF(2、10、50、250μg/(kg.d))，对照组以等量生理盐水进行处理。治疗1、3、5、7天后，进行行为学评分并取外周血计数单个核细胞，提取RNA 进行RT-PCR检测单个核细胞CD31、CXCR4 mRNA的表达水平；实验动物灌注固定后取脑组织切片检测G-CSF作用后脑组织中CD31及基质细胞衍生因子SDF-1表达。 结果 与生理盐水对照组相比，G-CSF治疗组神经功能评分较低（P0.05）；G-CSF 作用后外周血中单个核细胞数量明显增加；G-CSF低剂量作用组（2、10μg/kg）外周血单个核细胞CD31及CXCR4mRNA表达随着用药时间的延长而增强；高剂量（50、250μg/kg）在短时间内（1、3天），外周血单个核细胞CD31、CXCR4mRNA随着用药剂量的升高而增强，250μg/kg最强，且随着用药时间的延长（5、7天）， G-CSF作用逐渐减弱；免疫检测发现 G-CSF作用后脑组织内皮祖细胞标志CD31及SDF-1阳性表达明显增加。 结论 G-CSF能改善MCA0后大鼠症状，其机制可能在于动员骨髓干细胞进入外周血并促进脑组织SDF-1表达以趋化干细胞，继而促进脑组织的血管再生和神经修复。 关键词 卒中 G-CSF 动员 骨髓干细胞 血管再生 神经修复 中图分类号： R743 文献标识码：A Effect of G-CSF on neural restoration and mobilization of bone marrow stem cells in Focal Cerebral Ischemia mouse ZHANG Xingxiu1, GUO Huijuan1,Li Lin2, WANG Jian1, ZHENG Min＃2 (1 Department of the Second Affiliated Hospital, Chongqing Medical University，Chongqing 400010, China;2 Faculty of Basic Medical Sciences, Chongqing Medical University , Chongqing 400010, China) Abstract: ObjectiveTo investigate the effects and mechanism of Granulocyte-colony stimulating factor (G-CSF) on mobilization of bone marrow stem cells and on its recruitment to the infarct brain tissues of focal cerebral ischemia mouse. Methods: MCAO model was induced by using the filament occlusion method. 60 successful MCAO model rats were randomly divided into control group and G-CSF treated group. Treated group were subcutaneously injected with G-CSF ( 2,10,50,250 μg / (kg.d) ), for1,3,5,7days.Neurological scale were evaluated and the number of mononuclearcyte in peripheral blood were counted, and CD31 and CXCR4mRNA were detected by RT-PCR. Brain tissue were taken after perfusion fixation and sections were cut to check the expression of CD31 and SDF-1 by immunohi-stochemistry. Results：compared with the control group,