头皮位点药物注射对缺氧缺血性脑病新生大鼠内源性神经干细胞及Nogo-A表达的影响  

Effect of Scalp Site Medication Injection on Endogenous Neural Stem Cells and Expression of Nogo-A in Newborn Rats with Hypoxic-Ischemic Encephalopathy

作　　者：王春梅[1] 万国兰[1] 韩博[1] 

机构地区：[1]郑州大学第三附属医院小儿脑瘫康复治疗中心,郑州450052

出　　处：《实用儿科临床杂志》2010年第22期1744-1747,共4页Journal of Applied Clinical Pediatrics

摘　　要：目的研究头皮位点药物注射对HIE新生大鼠内源性神经干细胞增殖分化及神经再生抑制因子Nogo-A mRNA表达的影响。方法新生7日龄SD大鼠80只。随机分成4组:假手术组、模型组、位点注射9 g.L-1盐水组及位点注射V itB1、V itB12、组,每组各20只。采用结扎左侧颈总动脉,并吸入2 h氮氧混合气体制作新生大鼠HIE模型。位点注射9 g.L-1盐水组和位点注射V itB1、V itB12组在模型建立第14天,分别在大鼠的左侧额顶叶皮下注射等量9 g.L-1盐水或V itB1、V itB12混合稀释液,1次.d-1,共20 d;假手术组和模型组不予处理。各组于干预后7 d处死动物,40 g.L-1多聚甲醛心脏灌注后断头取脑组织,分别做连续冠状切片。用免疫荧光方法测定各组海马神经干细胞标志物巢蛋白(Nestin)表达,免疫组织化学方法检测其海马胶质纤维酸性蛋白(GFAP)和神经元特异性烯醇化酶(NSE)的表达,原位杂交法测定其海马神经再生抑制因子Nogo-A mRNA的表达水平。结果 1.位点注射可增加海马Nestin的表达,以注射V itB1、V itB12组表达最高,假手术组表达最少。2.海马NSE免疫组织化学显示假手术组和位点注射V itB1、V itB12组表达较多,模型组海马表达最少(P<0.05)。3.海马GFAP免疫组织化学显示假手术组表达最少,模型组最多,位点注射9 g.L-1盐水和位点注射V itB1、V itB12可抑制GFAP的表达,其中以位点注射V itB1、V itB12组抑制作用最为明显(P<0.05)。4.假手术组Nogo-A mRNA表达最少,模型组表达最高,位点注射9 g.L-1盐水组和位点注射V itB1、V itB12组表达水平均低于模型组,其中位点注射V itB1、V itB12组更为明显(P<0.05)。结论头皮位点药物注射可激发内源性神经干细胞产生,促进其增殖分化,同时可抑制神经再生抑制因子Nogo-A的表达,从而促进大脑神经的修复。Objective To investigate the effect of scalp site medication injection on endogenous neural stem cells proliferation,differen-tiation and the expression of the nerve regeneration inhibitory factor Nogo-A mRNA in newborn rats with hypoxic-ischemic encephalopathy（HIE）.Methods Seven-day-old SD rats were randomly divided into 4 groups： sham operation group（n = 20）,HIE model group（n=20） and sites injected with normal 9 g·L-1 saline group（n=20） and sites injected with VitB1 and VitB12 group（n=20）.Rat models of HIE were prepared by ligation of left common carotid artery and later with a temporary systemic hypoxia for 2 hours.Fourteen days later,sites injected with 9 g·L-1 saline group and the sites injected with VitB1 and VitB12 group were respectively injected with 9 g·L-1 saline and mixed diluent VitB1 and VitB12 in the left frontal lobes and the apieal lobe of the rats one time a day for 20 days.Sham operation group and the model group were not treated.After 7 days intervention,rats in each group were killed,brains were removed after cardiac perfusion organizations with 40 g·L-1 paraformaldehyde,and a series of slices were made separately.The expression of Nestin used as neural stem cell marker in hippocampus was determined by immunofluorescence,and the expression of glial fibrillary acidic protein（GFAP） and neuron specific enolase（NSE） were detected with immunohistochemisty.The expression of nerve regene-ration inhibitory factor Nogo-A mRNA was determined by in situ hybridization.Results 1.Site medication injection could increase the expression of Nestin in hippocampus,with the highest expression when injected with VitB1 and VitB12,but with the lowest expression in sham operation group.2.In sham operation group and sites injected with VitB1 and VitB12 group,NSE expressions were higher,while the model group had the least expression（P0.05）.3.The expressions of GFAP were least in hippocampus respectively in the sham operation and most in the model group by immunohistochemistry.Site

关 键 词：缺氧缺血性脑病 头皮位点药物注射 内源性神经干细胞 NOGO-A 

分 类 号：R722.1[医药卫生—儿科]