神经干细胞移植对发育期癫痫大鼠的影响  

Effects of transplantation of neural stem cells on developing rats with epilepsy

作　　者：李慧 戴园园 李蕊 刘娜 孙康钦 陈娇 程华 

机构地区：[1]徐州医科大学附属医院儿科,江苏省221000

出　　处：《江苏医药》2016年第11期1214-1217,F0002,共5页Jiangsu Medical Journal

基　　金：徐州市科技项目(XM12B023)

摘　　要：目的观察新生大鼠海马神经干细胞(NSCs)移植对发育期癫痫大鼠海马CA3区和齿状回苔藓纤维出芽(MFS)的影响。方法取新生24hSD大鼠海马,提取、培养及纯化NSCs并行免疫荧光染色鉴定。选择21日龄SD大鼠,建立戊四氮慢性癫痫点燃模型,分为无癫痫发作组(正常对照组)、癫痫未治疗组(癫痫组)、癫痫+PBS侧脑室注射组(假手术组)和癫痫+NSCs侧脑室注射组(治疗组),每组10只。在癫痫发作后第7天,假手术组和治疗组分别将PBS和5-嗅-2-脱氧尿苷(BrdU)标记的NSCs移植入大鼠侧脑室。术后8周,采用免疫组织化学方法观察海马CA3区和齿状回BrdU阳性细胞的变化,Timm银染色方法观察海马区MFS评分。结果培养的细胞聚集呈悬浮生长,形成典型的神经球,免疫荧光染色巢蛋白及BrdU呈阳性表达,并能不断分裂增殖,移植入癫痫大鼠海马区8周后能检测到其存在并向四周迁移。与正常对照组比较,癫痫组和假手术组大鼠MFS评分升高(P<0.05);与癫痫组和假手术组比较,治疗组MFS评分降低(P<0.05);癫痫组与假手术组MFS评分差异无统计学意义(P>0.05)。结论 NSCs移植入癫痫大鼠海马后能够存活,并且可以降低海马CA3区和齿状回MFS程度,有效抑制癫痫发作。Objective To observe the transplantation of neural stem cells（NSCs）on mossy fiber sprouting（MFS）in CA3 area of the developing rats with epilepsy.Methods The NSCs extracted,cultured and purified from the hippocampus of newborn SD rats at 24-hour after birth were identified with immunofluorescence staining.Thirty 21-day-old SD rats accepted continuous intraperitoneal injection of pentylenetetrazol to establish kindling model with epilepsy and equally randomized into 3groups of A（received no treatment）,B and C（transplanted with PBS and BrdUlabeled NSCs into the lateral ventricle respectively on the 7th day after the seizures）.Another ten21-day-old normal SD rats were taken as the controls（group D）.Immunohistochemical method and Timms silver staining method were used to observe the changes of BrdU-positive cells in CA3 area and dentate gyrus of the hippocampus and MFS situation in the hippocampus after 8 weeks,respectively.Results The suspension cultured cells gathered and formed typical neural spheres with nestin-and BrdU-positive expression showed by immunofluorescence staining.These cells had the abilities of self-renewal and pluripotent differentiation,which could be detected to move around after transplanted into the hippocampus of rats for 8 weeks.Compared with group D,MFS score was increased in groups of A and B（P〈0.05）.Compared with groups of A and B,MFS score was decreased in group C（P〈0.05）.There was no significant difference in MFS score between groups of A and B（P〉0.05）.Conclusion The NSCs transplanted into the rats with epilepsy can survive,which may reduce MFS in CA3 area and dentate gyrus of the hippocampus and effectively suppress epileptic seizure.

关 键 词：癫痫 神经干细胞移植 苔藓纤维出芽 

分 类 号：R742[医药卫生—神经病学与精神病学]