缺血性脑卒中大鼠模型骨髓干细胞的动员和神经修复作用  

Effects of G-CSF on nerve repair and mobilization of bone marrow stem cells in rat models of focal cerebral ischemic stroke

作　　者：张兴秀[1] 郭慧娟[1] 李琳 王健[1] 郑敏 

机构地区：[1]重庆医科大学附属第二医院神经内科,重庆400010 [2]庆医科大学基础医学院组织学与胚胎学教研室,重庆400016

出　　处：《第三军医大学学报》2012年第12期1192-1196,共5页Journal of Third Military Medical University

基　　金：国家自然科学基金(31100985);重庆市自然科学基金(CSTC2010BB5096)~~

摘　　要：目的研究粒细胞集落刺激因子(granulocyte-colony stimulating factor,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(middle cerebral artery occlusion,MCAO)模型,成功造模大鼠(60只)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 d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、CD133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治疗组神经功能有明显的改善(P<0.05);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.d)]外周血单个核细胞CD31及CXCR4 mRNA表达随着用药时间的延长而增强;高剂量[50、250μg/(kg.d)]在短时间内(1、3 d),外周血单个核细胞CD31、CXCR4 mRNA随着用药剂量的升高而增强,250μg/(kg.d)最强,且随着用药时间的延长(5、7 d),G-CSF作用逐渐减弱;G-CSF作用后脑组织内皮细胞标志CD31、CD133及SDF-1阳性表达明显增加。结论 G-CSF能改善MCAO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。Objective To investigate the effect and mechanism of granulocyte-colony stimulating factor（G-CSF） on mobilization of bone marrow stem cells,angiogenesis and nerve repair in infarct brain tissues of rats with focal cerebral ischemia.Methods The rat models of middle cerebral artery occlusion（MCAO） were constructed using filament occlusion method,and 60 MCAO model rats were randomly divided into a control group（n=12） and a G-CSF group（n=48）.The rats of the G-CSF group were subcutaneously injected with G-CSF [2,10,50 and 250 μg/（kg·d）] for 1,3,5 and 7 d,and the rats of the control group were injected with the same amount of normal saline.The neurological scale was evaluated.The number of mononuclear cells（MNCs） in peripheral blood was counted.The mRNA expressions of CD31 and CXCR4 were detected by RT-PCR.The expressions of CD31,CD133 and stromal cell-derived factor 1（SDF-1） were detected by immunohistochemistry.Results Compared with the control group,the neurological function of the rats was improved significantly（P〈0.05）,and the number of MNCs in peripheral blood increased significantly in the G-CSF group.The mRNA expression of CD31 and CXCR4 in MNCs increased in a time-dependent manner in the low-dose G-CSF groups [2 and 10 μg/（kg·d）].In the high-dose G-CSF groups [50 and 250 μg/（kg·d）],the mRNA expression of CD31 and CXCR4 in MNCs increased in a dose-dependent manner in the first 3 d,but the increase was gradually weakened from the 4th to the 7th day.The expression of CD31,CD133 and SDF-1 in the brain tissues of the G-CSF group significantly increased as compared with those of the control group.Conclusion The symptoms of MCAO model rats can be relieved by G-CSF,which can induce the mobilization of bone marrow stem cells in peripheral blood and upregulate the SDF-1 expression in brain tissues of MCAO model rats to promote angiogenesis and nerve repair.

关 键 词：卒中 G-CSF 动员 骨髓干细胞 血管再生 神经修复 

分 类 号：R332[医药卫生—人体生理学] R363.22[医药卫生—基础医学]