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目的探讨移植骨髓基质细胞(BMSCs)对永久性局灶性脑缺血大鼠脑室下区(SVZ)细胞增殖的影响及其增殖细胞类型分析。方法制作永久性局灶性脑缺血大鼠模型,分为空白对照组(MCAO)、PBS对照组(MCAO+PBS)和治疗组(MCAO+BMSCs),每组动物再分为脑缺血后7d和14d亚组。空白对照组不予处理,PBS对照组在脑缺血后1d移植PBS,治疗组在脑缺血后1d移植BMSCs。用Zausinger六分法检测神经功能恢复情况;注射5-溴脱氧鸟嘧啶核苷(BrdU)标记SVZ的新增殖细胞,免疫荧光双重标记检测新增殖细胞的类型。结果在脑缺血后7d和14d,与两个对照组相比,治疗组的神经功能评分较高,有显著性差异(P<0.05);脑缺血后14d,治疗组的大鼠患侧SVZ的BrdU阳性细胞较其余两组高,存在显著差异(P<0.05);免疫荧光双重标记显示,新增殖细胞表达神经元和星形胶质细胞的标志物,提示新增殖细胞为神经元和神经胶质细胞的混合体。结论BMSCs可以改善永久性局灶性脑缺血大鼠的神经功能,其机制可能与促进SVZ的神经细胞增殖有关。
Objective To investigate the effect of bone marrow stromal cells (BMSCs) transplantation on the proliferation of subventricular zone (SVZ) in permanent focal cerebral ischemia rats and the types of proliferating cells. Methods The model of permanent focal cerebral ischemia was made and divided into blank control group (MCAO), PBS control group (MCAO + PBS) and treatment group (MCAO + BMSCs) 7d and 14d subgroups. The blank control group was not treated. The PBS control group was transplanted with PBS 1 day after cerebral ischemia, and the treatment group was transplanted with BMSCs 1 day after cerebral ischemia. The recovery of neural function was detected by Zausinger’s six-point method. The newly proliferating cells labeled with BrdU-labeled SVZ were injected and double-labeled with immunofluorescence to detect the type of newly proliferating cells. Results Compared with the two control groups, the neurological scores of the treatment group were significantly higher at 7d and 14d after cerebral ischemia (P <0.05). At 14d after cerebral ischemia, the ipsilateral BrdU positive cells in SVZ were significantly higher than those in other two groups (P <0.05). Immunofluorescence double staining showed that newly proliferating cells expressed neuronal and astrocyte markers, suggesting that newly proliferating cells were neurons and A mixture of glial cells. Conclusion BMSCs can improve the neurological function of permanent focal cerebral ischemia in rats, which may be related to the promotion of proliferation of neural cells in SVZ.