中文摘要：

目的 探讨脑衰反应调节蛋白2（collapsin response mediator protein 2,CRMP2）对大鼠脑缺血再灌注损伤后神经元丢失及神经再生的影响及其可能的机制。方法 将60只成年雄性SD大鼠分成4组：假手术组（sham）、脑缺血/再灌注组（MCAO）、脑缺血＋质粒对照组（MCAO＋GFP）、脑缺血＋CRMP2真核表达质粒组（MCAO＋CRMP2）。将质粒注射入各组大鼠脑内,1 d后建立大鼠大脑中动脉缺血（middle cerebral artery occlusion,MCAO）/再灌注模型,术后5~7 d腹腔注射5-乙炔基-2＇脱氧尿嘧啶核苷（5-ethinyl deoxidization uracil nucleoside-2＇,Edu）,免疫荧光检测各组大鼠MAP2、GFAP、Edu标记细胞的阳性率及Nestin/Edu双标记阳性细胞的表达;采用Western blot检测CRMP2、脑源性神经营养因子（brain derived neurotrophic factor,BDNF）及N-甲基-D-天冬氨酸受体2B（N-methyl-D-aspartate receptor2B,NMDAR2B）的表达;免疫组化检测CRMP2的表达及其神经功能评分。结果 与sham组相比,脑缺血再灌损伤使MAP2表达降低（P〈0.05）,GFAP表达增高（P〈0.05）,同时促进了Edu标记阳性细胞[（62.00±7.65）、（62.60±7.06）]及Nestin/Edu标记阳性细胞[（31.60±5.50）、（31.60±5.40）]的表达（P〈0.05）。而CRMP2真核质粒干预之后降低了GFAP的表达（P〈0.05）,且升高了MAP2的表达（P〈0.05）,还进一步促进了Edu标记阳性细胞（100.20±11.52）及Nestin/Edu标记阳性细胞（57.20±4.80）的表达（P〈0.05）。与sham组相比,MCAO及MCAO＋GFP组BDNF及NMDAR2B的表达明显升高（P〈0.05）;经CRMP2真核质粒干预后,BDNF表达进一步增高（P〈0.05）,NMDAR2B的表达被部分削弱（P〈0.05）。CRMP2真核质粒干预后神经功能评分较MCAO及MCAO＋GFP组明显改善（P〈0.05）。结论 CRMP2减少了大鼠脑缺血再灌注损伤后神经元的丢失;促进了大鼠脑缺血再灌注损伤后内源性神经干细胞的增殖,而其对BDNF及NMDAR2B的调节可能是其作用机?

英文摘要：

Objective To determine the effect of collapsin response mediator protein 2 （CRMP2） on the neuronal loss and neurogenesis in the rats after cerebral ischemia/reperfusion （I/R） injury and investigate its possible mechanisms. Methods A total of 60 adult male SD rats were divided into 4 groups, that is, sham operation group, cerebral I/R group （middle cerebral artery occlusion, MCAO）, cerebral ischemia with blank plasmid control group （MCAO ＋ GFP group ）, and cerebral ischemia with CRMP2 eukaryotic plasmid group （MCAO ＋ CRMP2 group）. In 1 d after the injection of corresponding plasmids, the cerebral I/R model was established. In 5 to 7 d after the operation, 5-ethinyl deoxidization uracil nucleoside- 2＇ （Edu） was injected intraperitoneally into these rats. Then the expression of MAP2, GFAP, Edu and Nestin/Edu was tested by immunof[uorescenee assay. The expression levels of CRMP2, brain derived neurotrophic factor （BDNF） and N-methyl-D-aspartate receptor 2B （NMDAR2B） were detected by Western blotting. The location of CRMP2 was tested by immunohistochemical staining. Neurological function was also evaluated. Results Compared with the sham group, cerebral I/R injury decreased the expression level of MAP2 （P 〈 0.05 ）, while increased that of GFAP （ P 〈 0.05 ）, as well as the numbers of Edu （ 62.00 ± 7.65, 62.60 ± 7.06） and Edu/Nestin （31.6 ± 5.50, 31.6 ± 5.40） positive cells in the MCAO and MCAO ＋ GFP groups （ P 〈 0. 05 ）. Intervention of CRMP2 eukaryotic plasmid increased the expression of MAP2 （ P 〈 0.05 ）, inhibited the expression of GFAP （ P 〈 0.05 ）, and boosted the amounts of Edu （ 100.20± 11.52 ） and Nestin/Edu （57.20± 4.80） positive cells （ P 〈 0.05 ）. Compared with the sham group, the expression levels of BDNF and NMDAR2B were obviously increased in the MCAO and MCAO ＋ GFP groups （P 〈 0. 05）. After the intervention of CRMP2 eukaryotic plasmid, the expression of BDNF was further enhanced （P 〈 0.05 ）, an