转Nurr1基因联合全反式维甲酸及银杏提取物诱导的神经干细胞治疗帕金森病大鼠模型  

An experimental study of therapy to rat model of Parkinson's disease by transplantation of neural stem cells modified by Nurr1 gene and induced by all-trans retinoic acid /ginkgo biloba in vitro

作　　者：李小元[1] 陈先文[1] 徐仿成[1] 胡慧敏[1] 王烈成[1] 

机构地区：[1]安徽医科大学第一附属医院神经内科,合肥230022

出　　处：《安徽医科大学学报》2007年第5期520-524,共5页Acta Universitatis Medicinalis Anhui

基　　金：安徽省自然科学基金资助项目(编号:01043401)

摘　　要：目的探讨核受体相关因子(Nurr1)基因修饰联合全反式维甲酸(ATRA)及银杏提取物(Egb)诱导的神经干细胞(NSCs)移植治疗帕金森病动物模型的疗效及作用机制。方法将建模成功的帕金森病SD大鼠模型分为4组,将溴尿嘧啶(Brdu)标记的单纯胚胎NSCs(NSCs组)、RA/Egb诱导分化的NSCs(RA/Egb组)、单纯转Nurr1基因的NSCs(Nurr1组)和转Nurr1基因联合RA/Egb诱导分化的NSCs(Nurr1+RA/Egb组)分别注入模型鼠右侧纹状体。观察其病理性旋转行为的改善,采用免疫组化染色方法检测酪氨酸羟化酶(TH)、Nurr1、神经丝巢蛋白(nestin)表达及Brdu标记情况,观察移植的NSCs在体内的存活、迁移与分化,并进行TH阳性细胞计数研究脑内多巴胺含量的变化。结果各组动物脑内移植后动物旋转行为较前均有改善,尤以Nurr1+RA/Egb组改善最明显,RA/Egb组次之。免疫组化可见各组移植区均有TH阳性细胞表达,其中Nurr1+RA/Egb组TH染色细胞数更密集,形态更成熟,病理学改善最明显。在Nurr1组和Nurr1+RA/Egb组移植区可见有Nurr1散在表达,在NSCs组和RA/Egb组移植区未见Nurr1明显表达。nestin检测未见有明显阳性表达。结论转Nurr1基因联合全反式维甲酸及银杏提取物诱导的神经干细胞脑内移植后可以存活、迁移、分化并表达TH和Nurr1,部分地改善PD模型旋转行为,增加脑内TH表达。Objective To observe curative effects and mechanism of neural stem cells modified by Nurrl gene and induced by all-trans retinoic acid/ginkgo biloba （RA/Egb） in vitro transplantation on rat Parkinson＇disease（PD） model. Methods The 6-hydroxydopamine（6-OHDA）-induced hemiparkinsonian rats were seleceted and allocated randomly into 4 groups. NSCs with brdu-labeled （2× 10^5 cells/each model） （ NSCs group）, NSCs induced by RA/ Egb（ RA/Egb group）, NSCs modified by Nurrl gene （Nurrl group）, NSCs modified and induced by RA/Egb （ Nurrl ＋ RA/Egb group） were injected into right striatum of PD rat respectively. The rotation test was taken from the first week to the twelfth week after transplantation. Brain paraffin sections were carried out immunohistochemical staining （ brdu, nestin, Nurrl and TH） afer one week, two week, six week, ten week, one, two, three month of NSCs transplantation, meanwhile determining the survival ,immigration and differentiation of NSCs in the brain of PD model afer transplantion and counting the number of TH-positive cells to study the change of DA number in the brain. Results The rotation of rats in all groups revealed decreased after transplantation, especially in Nurrl ＋ RA/Egb group, the improvement of rotation in RA/Egb group was less. In immunohistochemical staining some TH-positive cells could be found in the implanted region of all groups. In Nurrl ＋ RA/Egb group, The number of TH-positive cells increased obviously, the dendrite and neurite of TH-positive cells was more intensive, and the pathological improvement was the best. Some Nurrl-positive was found in the implanted region of Nurrl group and Nurrl ＋ RA/ Egb group. Meanwhile nestin-positive could not was found evidently in the implanted region. Conclusion Neural stem cells modified by Nurrl gene and induced by all-trans retinoic acid/ginkgo biloba in vitro can survive, immigration , differentiate and express TH, Nurrl in the brain of PD model after transplantation, and can improve the rot

关 键 词：干细胞 神经系统 基因疗法 银杏/药物作用 维甲酸/药理学 帕金森病/中药疗法 植物提取物/治疗应用 大鼠 

分 类 号：R617[医药卫生—外科学] R394.26[医药卫生—临床医学]