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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 after intracerebral hemorrhage (ICH) in rats. Methods Ninety SD male rats were randomly divided into three groups: TPC group, ICH group and control group. Each group was divided into subgroup 3,7,14,21,28 d; stereotactic injection of type IV collagenase brain intracerebral hemorrhage model was made, the new SVZ cells were labeled with Brdu; brain slices were cultured, the time interval microscopy The migration of Dil-labeled SVZ cells in living brain slices was dynamically observed. To evaluate whether SVZ cells migrated to the injured area had the ability to form synapses with other cells, double-stained immunohistochemical staining of Brdu and synaptic protein Ⅰ was performed. Results The migration of Dil-labeled SVZ cells to the striatum was observed under the microscopic time-interval microscope. The velocity of SVZ cells migration was (4.23 ± 0.25) μm / h at 3 d in TPC group and (8.23 ± 0.47) μm / h at 14 days, (7.05 ± 0.31) μm / h at 21 days and (5.29 ± 0.20) μm / At each time point, the TPC group migrated faster than the cerebral hemorrhage group (P <0.01). TPC group of ipsilateral striatum Brdu and synuclein double positive double positive cells increased significantly 3 d, reached its peak on the 14 d, continued until 21 d, and then gradually decreased. BrdU and synapse Ⅰ double positive cells were not seen in 3 days after intracerebral hemorrhage. A few positive cells were seen on day 7 and reached a peak on day 14, then decreased gradually. At each time point, double positive cells were significantly increased compared with intracerebral hemorrhage (P <0.01). TPC allows the expression of synapsin I occurs earlier, longer duration. Conclusion TPC can promote the migration and differentiation of SVZ cells after intracerebral hemorrhage.