凝血酶预处理对大鼠脑出血后SVZ细胞迁移和分化的影响  

The Effect of thrombin preconditioning on migration and differentiation of subventricular zone cells after intracerebral hemorrhage in rats

作　　者：关景霞[1] 解燕春[1] 张少锋[1] 袁振华[1] 黄婷婷[1] 卢祖能[1] 

机构地区：[1]武汉大学人民医院神经内科,430060

出　　处：《卒中与神经疾病》2016年第4期232-236,共5页Stroke and Nervous Diseases

基　　金：国家自然科学基金资助项目(项目批准号为81301010)

摘　　要：目的探讨凝血酶预处理(thrombin preconditioning,TPC)对大鼠脑出血(intracerebral hemorrhage,ICH)后侧脑室下带(subventricular zone,SVZ)细胞迁移和分化的影响。方法将90只SD雄性大鼠随机分为3组:TPC组、ICH组、对照组。每组分为3、7、14、21、28 d亚组;应用IV型胶原酶脑内立体定向注射制作脑出血模型,应用Brdu标记新生的SVZ细胞;进行脑片培养,应用时间间隔显微系统动态观察Dil标记的SVZ细胞在活体脑片上的迁移;为了评估迁移至损伤区的SVZ细胞是否具有与其他细胞形成突触的能力,进行Brdu和突触蛋白Ⅰ免疫组织化学双标染色。结果在时间间隔显微系统下观察Dil标记的SVZ细胞向纹状体迁移,动态观察12 h,TPC组3 d SVZ细胞迁移的速度是(4.23±0.25)μm/h,7 d的速度是(6.53±0.37)μm/h,14 d的速度是(8.23±0.47)μm/h,21 d的速度是(7.05±0.31)μm/h,28 d的速度是(5.29±0.20)μm/h,在各个时间点TPC组的迁移速度明显快于脑出血组(P<0.01)。TPC组同侧纹状体Brdu和突触蛋白Ⅰ双标阳性细胞3 d明显增加,14 d达高峰,持续至21 d,然后逐渐减少。脑出血组3 d未见Brdu和突触蛋白Ⅰ双标阳性细胞,7 d可见少数阳性细胞,14 d达高峰,然后逐渐下降,在各个时间点双标阳性细胞与脑出血组比较均明显增加(P<0.01)。TPC使突触蛋白I的表达出现时间更早,持续时间更长。结论 TPC能够促进脑出血后SVZ细胞的迁移和分化。Objective To investigate the effect of thrombin preconditioning （TPC） on the migration and differentiation of subventricular zone （SVZ） cells in rats after intracerebral hemorrhage （ICH）. Methods 90 Male Sprague-Dawley rats were randomly divided into 3 groups （ICH, TPC and control group）. Rats of each group were randomly divided into 5 subgroups （3 d, 7 d, 14 d, 21 d, 28 d subgroup）. ICH was caused by intrastrial stereotactic administration of collagenase type IV. Brdu was used to label newborn SVZ cells. To dynamically observe the migration of SVZ cells at living brain tissue, brain slices were cultured. Migration of Dillabeled SVZ cells in living brain slice was traced by timeqapse microscopy. To assess whether SVZ cells migrating to injured striatum had the ability to form synapses with other cells, the brain slices from each group were double inu-nunolabeled with Brdu and synapsin I. Results Migration of Dil-labeled SVZ cells to the striatum in brain slices in each group was dynamically observed and imaged by timeqapse microscopy during a 12 hour pe- riod. The velocity of migration of Dil-labeled SVZ cells in the TPC group was 4. 23 ± 0. 25 m/h in the 3-day group, 6. 53± 0. 37 m/h in the 7-day group, 8. 23± 0. 47 m/h in the 14-day group, 7.05± 0. 31 m/h in the 21 day group, and 5.29 ± 0. 20 m/h in the 28-day group respectively. The velocity of migration in the TPC group was faster than that in the ICH group with significant statistic difference （P^0. 01）. The numbers of Brdu and synapsin positive cells were markedly increased in the ipsilateral striatum at 3 days after TPC, peaked at 14 days （P〈0. 01）, continued to 21 days, and then gradually decreased at 28 days. At 3 days after ICH, all of the Brdu-positive cells in the ipsilateral stratum were not synapsin I -positive cells. At 7 days after ICH, the minority of Brdu-positive cells were synapsin I -positive cells. Colocalization of Brdu with synapsin I peaked at 14 days and then gradually decreased. There was significant s

关 键 词：脑出血 侧脑室下带 凝血酶预处理 

分 类 号：R743.34[医药卫生—神经病学与精神病学]