骨髓基质细胞侧脑室移植对创伤性颅脑损伤鼠认知功能的影响

The influence of bone marrow stromal cells transplantation into lateral ventricle on the cognitive function of traumatic brain injury rats

目的探讨骨髓基质细胞(bone marrow stromal cells,BMSC)移植对创伤性颅脑损伤(traumatic brain injury,TBI)大鼠认知功能的影响。方法采用Feeney自由落体方法成功制作72只TBI模型大鼠,随机分为BMSC组、TBI组、IMDM组,每组24只,取24只正常大鼠作正常对照组。原代培养带绿色荧光蛋白(green fluorescent protein,GFP)的BMSC并移植到BMSC组模型大鼠的侧脑室,IMDM组大鼠侧脑室注入等体积IMDM培养基,TBI组作空白对照组。4组于移植后第2、4、8、12周行Morris水迷宫实验,取BMSC组大鼠脑行冰冻切片及免疫荧光染色,观察移植细胞的存活、迁移和分化。结果在实验的各个时间点,与正常对照组相比较,TBI组平均逃避潜伏期显著延长,穿过原平台位置的次数显著变少(均P=0.000);移植4周后,与IMDM组、TBI组相比较,BMSC组平均逃避潜伏期显著缩短(均P<0.05),穿过原平台位置的次数显著增多(均P<0.05)。免疫荧光结果显示:大量BMSC迁移到损伤的皮质区域,部分细胞能表达胶质纤维酸性蛋白(GFAP)和神经元核心抗原(NeuN)的表型。结论 TBI可损害大鼠的认知功能,将BMSC移植到TBI大鼠的侧脑室,移植细胞能迁移到损伤的皮质区域,部分细胞分化为神经元和神经胶质细胞,且能改善大鼠的认知功能。

Objective To explore the influence of bone marrow stromal cells （BMSC） transplantation into the lateral ventricle on the cognitive function of experimental traumatic brain injury （TBI） rats. Methods Seventy-two experimental TBI model rats were successfully established according to the method of Feeney t^eely falling object, which were randomly and equally divided into BMSC group, TBI group and IMDM group. Primary BMSC with green fluorescent protein （GFP） were cultured and transplanted into ventricles of TBI rats in BMSC group. In IMDM group, rats received equal volume of IMDM culture medium. TBI group was set as blank control group and 24 healthy rats as normal control group. Morris water maze test was performed 2, 4, 8 and 12 weeks after the transplantation. Frozen sections of brain tissue of rats in BMSC group were prepared and immunofluorescent staining was used to detect the migration and differentiation of BMSC. Results Compared with the normal control group, the mean escape latency was longer and the times through platform quadrant were less at each time point of experiment （all P=-0.000） in TBI group. Compared with IMDM and TBI groups, the mean escape latency was shorter and the times through platform quadrant were more in BMSC group 4 weeks after transplantation （all P〈0.05）. And lots of BMSC migrated to the injured cortical area, and part of ceils expressed glial flbrillary acidic protein （GFAP） and NeuN phenotype, which were observed under fluorescence microscope. Conclusions TBI can damage cognitive fimction of rats. After BMSC transplantation into the lateral ventricle of TBI rats, BMSC can not only migrate to the injured cortical area and part of cells differentiate into neurons and glial cells, but also improve the cognitive function of rats.