经枕大池移植神经干细胞治疗重型颅脑损伤大鼠的实验研究

The rapeutic study of neural stem cells injected into cistern magna on the rat model of severe craniocerebral injury

目的探讨胎脑皮质来源的神经干细胞(NSC)用于重型颅脑损伤模型大鼠的治疗作用。方法用电子颅脑损伤仪制备重型颅脑损伤大鼠模型(n=46),随即分为神经干细胞治疗组(NSC组,n=23)和对照组(DMEM/F12组,n=23)。将胎鼠皮质来源的NSC经体外培养、扩增后用BrdU标记,经枕大池移植到模型大鼠的脑脊液中,于移植后24h,3d,7d,14d,21d,28d对大鼠进行改良神经功能缺损评分(mNSS)的评估。对照组用相同体积的DMEM/F12代替NSC,mNSS评估时间和方法同NSC组。4w后NSC组取5只大鼠脑组织行免疫组化法染色检测BrdU表达。结果 NSC组大鼠在移植后第7d、14d、21d和24d,其mNSS评分与DMEM/F12组相比,均有显著性差异。对脑组织行BrdU免疫荧光染色,发现移植后第4w脑损伤灶及周围脑组织中有BrdU阳性细胞的表达。结论经枕大池移植后,NSC可在大鼠脑组织内存活、增殖,并能促进脑损伤大鼠神经功能的恢复。

Objective To observe the effect of fetal brain cortex derived neural stem cells transplantation on treating severe craniocerebral injury rats. Method 46 severe craniocerebral injury rats were generated by electronic craniocerebral injury instrument, and randomly divided into the neural stem cell therapy group （n = 23 ） and the control group （n = 23 ）. After culturing for 3 days, the third generation passage cultured neural stem cells were labeled with BrdU, and injected into the occipital pool. The control group were injected with same volume of DMEM/F12. Two groups of rats were tested improved neurological deficit scores after injection on the 24 hours,3^rd, 7^th, 14^th, 21^st and 24^th day. Four weeks after transplantation, five rats of the therapy group were sacrificed, and the BrdU-positive cells in brain were examined by immunohistochemical method. Result The results showed that after transplantation of neural stem cells, on the 7^th, 14^th, 21^th and 24^th day, the paralysis symptoms were greatly improved. The neurological deficit score of the therapy group were significantly different compared with the control group. A large number of BrdU-positive ceils were observed around the foci of brain damage in the 5 sacrificed rats after four weeks transplantation. Conclusion Transplantation of cultured neural stem cells can remain alive and grow in the rat brain, and facilitate the functional recovery in craniocerebral injury rat.