人胚神经干细胞移植对大鼠脑液压冲击伤的影响

The effects of human embryonic neural stem cells on rat brain fluid percussion injury

目的 通过人胚神经干细胞 (HNSCs)移植大鼠脑液压冲击伤 (FPI)探讨HNSCs对脑损伤修复的影响。方法 体外培养HNSCs并用 5 溴脱氧尿嘧啶 (BrdU)标记 ,液压冲击大鼠右侧大脑皮质运动感觉区制作脑外伤模型 ,脑外伤后 2 4h移植HNSCs ,分别于脑外伤前、脑外伤后 2 4h、4周行神经运动行为学评分 (NMFE)和检测左下肢的短潜伏期体感诱发电位 (SLSEP)及免疫组织化学染色检测BrdU、巢蛋白 (nestin)、微管相关蛋白 2 (MAP2 )、胶质纤维酸性蛋白 (GFAP)和半乳糖脑苷 (GalC)蛋白表达。结果 大鼠液压冲击伤后 2 4h ,左下肢功能障碍明显 ,运动评分14 .3 75 0± 2 .13 3 9,SLSEP潜伏期 3 0 .463± 4.64 0与损伤前相比均差异有显著性 (P <0 .0 1) ,但移植HNSCs组与对照组差异无显著性 ;HNSCs移植后 1周 ,BrdU阳性细胞从移植区域向周围扩散 ,邻片nestin、MAP2和GFAP均为阳性 ,但MAP2阳性细胞较多 ;HNSCs移植后 4周 ,未见nestin阳性细胞 ,液压冲击区域聚集GFAP/BrdU双阳性细胞和MAP2 /BrdU双阳性细胞 ,且GFAP/BrdU阳性细胞较多。结论 HNSCs移植对大鼠脑液压冲击伤的功能恢复无影响 ,但早期HNSCs多数分化为神经细胞 ,晚期多数分化为神经胶质细胞。

Objective To investigate the effects of human embryonic neural stem cells (HNSCs) on the repair of rat brain fluid percussion injury.Methods HNSCs were cultured in vitro,labeled with BrdU,and transplanted into rat brain area injured by fluid percussion.The short-latency somatosensory evoked potential (SLSEP) and neurological motor function evaluation (NMFE) were used to test the functions at pre-injury and 24 h as well as 4 weeks post-injury.Immunohistochemistry was used to detect the expression of BrdU,nestin,MAP2,GFAP and GalC.Results There was significant difference in SLSEP (30.463±4.640) and NMFE (14.375?0±2.133?9) between pre-injury and 24 h post-injury ( P < 0.01),but there was no significant difference between HNSCs transplantation group and control group.One week after HNSCs transplantation,BrdU positive cells could be seen and the expression of nestin,MAP2 and GFAP was detectable simultaneously in the same positon using immunohistochemical staining,but the number of MAP2 positive cells was more than GFAP.Four weeks after transplantation,GFAP positive cells were more than MAP2 and the expression of nestin was not detectable.Conclusion HNSCs have no effects on rat traumatic brain injury.HNSCs can differentiate into neurons at early stage of brain injury and into astrocytes at late stage.