GDNF及dbcAMP对神经干细胞诱导分化为多巴胺能神经元作用的实验研究  

Experimental study of the functions of GDNF and dbcAMP on the induction of differentiation of neural stem cells into dopaminergic neurons

作　　者：陈立[1,2] 杜娟[3] 陈丽[1] 吕晓红[1] 

机构地区：[1]吉林大学白求恩第一医院神经内科,吉林长春130021 [2]华中科技大学同济医学院附属荆州医院神经内科 [3]烟台市烟台山医院神经内科,山东烟台264000

出　　处：《中风与神经疾病杂志》2014年第7期580-584,共5页Journal of Apoplexy and Nervous Diseases

摘　　要：目的 利用GDNF及dbcAMP对大鼠胚胎神经干细胞进行定向分化,从而获得相当数量的多巴胺能细胞,为下一步的细胞移植打下基础。方法 分离培养孕14~16 d Wistar大鼠大脑神经干细胞,通过传代培养来检测其自我更新能力,并对其进行Nestin及Brdu蛋白鉴定,利用GDNF及dbcAMP对大鼠神经干细胞进行定向分化,通过免疫细胞化学及免疫荧光等方法检测TH阳性细胞表达。结果 培养的神经干细胞均表达Nestin及Brdu,体外培养1 w后神经球的数量显著增加,体积增大;GDNF及dbcAMP均能提高TH表达,与血清对照组比较有显著差异（P〈0.05）。结论 在神经营养因子的作用下,分离的神经干细胞能够快速增殖并形成神经球,GDNF及dbcAMP均有诱导神经干细胞向多巴胺能神经元分化的作用,其中GDNF在20μg/L时诱导效能最高,dbcAMP在200μmol/L时诱导效能最高,二者均存在剂量依赖性,二者联合诱导可明显提高各自单独诱导TH细胞的阳性率。Objective To obtain a considerable number of dopaminergic cells through the use of GDNF and db- cAMP on rat embryonic neural stem cells for directed differentiation and therefore lay the foundation for the next cell trans- plantation. Methods Neural stem ceils of 14-16 days pregnant Wistar rat were isolated and cultured, cells were subcul- tured to detect its self-renewal capacity and its Nestin and Brdu protein, identification, and then take use of rat neural stem cells that differentiation directed by GDNF and dbcAMP,the expression of TH positive cells were detected by immunocyto- chemistry and immunofluorescence methods. Results Cultured neural stem cells expressed Nestin and Brdu. A significant increase in the number of neurospheres after one week of in vitro culture were observed, the volume increased too. GDNF and dbeAMP could increase TH expression respectively and there were significant differences compared with the serum con- trol group（ P 〈 0.05 ）. Conclusion In the role of the neurotrophic factor, neural stem cells can rapidly proliferate and form neurospheres. Both GDNF and dbcAMP can result in the induction of differentiation of neural stem cells into dopaminergie neurons, GDNF induces the highest performance in 20 μg/L and dbcAMP induces the highest performance in 200 ~mol/L, both in a does-dependent manner. There is significant difference in TH cells positive rate between combination of induction and independent induction.

关 键 词：神经干细胞 定向分化 多巴胺能神经元 GDNF DBCAMP 

分 类 号：R742.5[医药卫生—神经病学与精神病学]