神经节苷脂联合神经干细胞移植对颅脑损伤大鼠海马的作用

Effects of ganglioside combined with neural stem cells transplantation on hippocampal neuron following brain injury in rats

背景:应用神经干细胞移植治疗脑损伤后神经功能障碍的实验研究是目前的热点。目的:观察神经节苷脂联合神经干细胞移植对创伤性脑损伤大鼠海马神经元的影响。方法:健康Wistar大鼠66只,采用改进Feeney法制作创伤性脑损伤致海马神经元损伤模型,存活的60只大鼠伤后24h给予相应治疗随机分为3组:创伤性脑损伤组注射1mL DMEM/F12培养液;神经干细胞组注射1×1010L-1神经干细胞悬液;神经干细胞+神经节苷脂组注射1×1010L-1神经干细胞悬液后经腹腔注射30mg/kg神经节苷脂水溶液,1次/d,连续3d。结果与结论:荧光显微镜观察PKH26标记的神经干细胞呈球形,呈现均匀分布的红色荧光;PKH26标记神经干细胞的阳性率为95%。各组平均潜伏时间均逐渐缩短,神经干细胞+神经节苷脂组3～5d时平均潜伏时间较神经干细胞组缩短(P<0.05),较创伤性脑损伤组明显缩短(P<0.01);神经干细胞+神经节苷脂组穿越平台次数及在目标象限游泳距离与总距离百分比均高于其他两组(P<0.05)。PKH26阳性细胞和苏木精-伊红染色切片中的神经元数量及脑源性神经营养因子mRNA的表达神经干细胞+神经节苷脂组多于神经干细胞组,神经干细胞组多于创伤性脑损伤组(P<0.05)。提示神经干细胞移植对创伤性脑损伤大鼠海马神经元有保护作用,联合应用神经节苷脂有协同效果。

BACKGROUND：The research about the application of neural stem cells（NSCs） transplantation on the treatment of neurological dysfunction after brain injury is a hot spot.OBJECTIVE：To investigate the effect of ganglioside combined with NSCs transplantation on hippocanipal neuron following traumatic brain injury（TBI） in rats.METHODS：Sixty-six healthy Wistar rats were selected to establish the hippocanipal neuron model cause by TBI using improved Feeney method.After injured for 24 hours,the surviving 60 rats were randomly divided into 3 groups,the TBI group was injected with 1 mL DMEM/F12 medium;the NSCs group was injected with 1×1010/L NSCs suspension;the NSCs＋ganglioside group was injected with 1×1010/L NSCs suspension and then intraperitoneal injection of 30 mg/kg ganglioside aqueous solution,once per day and lasted for 3 days.RESULTS AND CONCLUSION：Fluorescence microscope observation showed that the NSCs labeled by PKH26 were spherical and presented with red and uniform-distributed fluorescence.Flow cytometric detection revealed that the positive percentage of PKH26 labeled NSCs was over 95%.The average time of escape latency was gradually decreased in each group.However,from the 3rd to 5th day,time in NSCs＋ganglioside group was shorter than that in NSCs transplantation group（P 0.05） and compared with TBI group,time in NSCs＋ganglioside group was decreased significantly（P 0.01）.In addition,the frequency of platform passing and the percentage of swimming distance in target quadrant and the total distance were highest in NSCs＋ganglioside group（P 0.05）.The number of PKH26-positive cells and neurons in the section stained by hematoxylin-eosin as well as the mRNA expression of brain-derived neurotrophic factor in NSCs＋ganglioside group was more than that in NSCs group,and those factors in NSCs group were more than those in TBI group（P 0.05）.NSCs transplantation plays a role in protecting against hippocampal neuronal death following TBI,with the affiliation of ganglioside claim synergism effects.