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为了探讨切割海马伞后的海马和正常海马在蛋白表达方面的差异,获取海马中具有生物活性的特异蛋白,并观察其对人胚神经干细胞迁移的影响,取切割SD大鼠海马伞后10、14、20d及正常海马进行非变性聚丙烯酰胺凝胶电泳(NativePAGE)。将用无血清培养技术获取的人胚神经干细胞球分别与切割海马伞后14d海马和正常海马的56kD差异蛋白胶条及空白对照胶条共培养,观察神经干细胞球中神经干细胞迁移情况,第3d时计数朝胶条方向1/4扇区内的迁出细胞数。结果显示,切割后各天组和正常组比较见多条差异蛋白条带,其中56kD差异蛋白的密度积分正常组最低,10d组和20d组较高,14d组最高。神经干细胞球中向56kD差异蛋白胶条方向迁移的细胞数在切割组最多,正常组次之,对照组最少。提示切割海马伞后的海马与正常海马间存在多种差异蛋白,其中56kD蛋白在切割海马伞后14d表达量最高,并具有诱导神经干细胞迁移的作用。
To investigate the difference in protein expression between hippocampus and normal hippocampus after cutting the hippocampus, to obtain the specific protein with biological activity in hippocampus and to observe its effect on the migration of human embryonic neural stem cells. After cutting the hippocampus of SD rats 10, 14,20 and normal hippocampal non-denaturing polyacrylamide gel electrophoresis (NativePAGE). Cultured human embryonic neural stem cells obtained with serum-free culture technique were co-cultured with 56 kD differentially expressed protein strips and blank control strips in hippocampus and normal hippocampus 14 days after cutting the hippocampus to observe the migration of neural stem cells in the NSCs. Count the number of migrating cells within 1/4 sector of the strip. The results showed that there were many bands of different proteins in each group after cutting. The 56kD differential protein was the lowest in the normal group, the highest in the 10d group and the 20d group, and the highest in the 14d group. The number of cells migrating in the direction of the 56 kD differential protein gel in the NSCs was the highest in the cutting group, the normal group followed by the least in the control group. The results suggest that there are many different proteins between the hippocampus and the normal hippocampus after cutting the hippocampus. The 56kD protein has the highest expression level at 14d after cutting the hippocampus and induces the migration of neural stem cells.