Neurturin基因转染c17·2神经干细胞移植保护帕金森病大鼠模型的实验研究  

Experimental study of neuroprotection through delivery of neurturin by c17.2 neural stem cells in a rat model of Parkinson’s disease

作　　者：刘卫国[1,2] 陈生弟[1] 陆国强[1] 李彪[1] 王宝奇[1] 陈琰[1] 杜芸兰[1] 

机构地区：[1]上海交通大学附属瑞金医院神经科,帕金森病诊疗与研究中心,现在南京脑科医院神经内科200025 [2]南京脑科医院神经内科

出　　处：《中华神经科杂志》2006年第3期184-188,共5页Chinese Journal of Neurology

基　　金：国家重点基础研究规划资助项目(G1999054008);国家自然科学基金资助项目(39970263;30171025)

摘　　要：目的探讨neurturin(NTN)基因转染c17·2神经干细胞脑内移植后对帕金森病大鼠模型的保护作用。方法构建高表达NTN蛋白的NTN-c17·2细胞克隆。实验分成NTN(12只)、空质粒(Mock,9只)和PBS组(9只),分别在各组大鼠的纹状体区移植入NTN-c17·2、Mock-c17·2和PBS,16d后再注入6-羟多巴胺(6-OHDA)。通过免疫组化、原位杂交、旋转行为和神经递质变化观察c17·2神经干细胞的分化,NTN基因的表达和NTN蛋白的保护作用。结果NTN-c17·2细胞移植后,细胞分化但仍持续分泌NTN蛋白。NTN蛋白可逆向运输到黑质,保护酪氨酸羟化酶(TH)阳性神经元免受6-OHDA的损害。动物模型旋转行为动态观测显示NTN的保护作用至少持续了4个月,NTN组要明显好于Mock和PBS组。移植后10个月,NTN和Mock组动物移植侧与健侧纹状体的神经递质如多巴胺浓度比值分别为95%(P<0·05)和93%(P<0·05),PBS组为75%。结论用带有NTN基因的神经干细胞移植治疗帕金森病,显示出良好的保护效果。Objective To explore the neuroprotection through delivery of neurturin by c17. 2 neural stem cells in a rat model of Parkinson＇ s disease. Methods Constructing c17. 2-NTN clone overexpressed NTN protein. Injecting NTN-c17.2, Mock-c17.2 and PBS into the striatum of SD rats in NTN Mock and PBS groups respectively, 6-OHDA injection 16 days later. Observing the differentiation of c17.2 neural stem cells, the expression and neuroprotection of NTN by using immunohistochemical staining, in suit hybridization, rotational behavior and HPLC respectively. Results After intrastriatal grafting NTN-cl7.2, grafts stably overexpressed NTN. Intrastriatal grafting of the NTN-cl7.2 resulted in the retrograde labeling of substantia nigra dopaminergic neurons, which protected them from the toxicity of 6-OHDA. Circling behavior indicted the protective effect of NTN-cl7.2 lasting at least for 4 months and the animals receiving NTN- c17. 2 grafts plus 6-OHDA showing obvious reduction in the number of net contralateral rotations compared with the animals receiving PBS plus 6-OHDA or Mock-el7.2 plus 6-OHDA. HPLC tests indicated that the contents of neurotransmitters （e. g. dopamine） in the corpus striatum area of the NTN-cl7. 2 group（95%, P〈0.05） and the Mock-el7. 2 （93% ,P 〈0.05） group were significantly higher than in the PBS group （75%）. Conclusions The transplantation of NTN-cl7.2 exerted protective effects in rat model of PD.

关 键 词：干细胞移植 神经组织蛋白质类 帕金森病 转染 大鼠 

分 类 号：R742.5[医药卫生—神经病学与精神病学]