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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 investigate the protective effect of neurturin (NTN) gene transfected c17 · 2 neural stem cells on Parkinson’s disease rat model after intracerebral transplantation. Methods NTN-c17.2 cell clone with high expression of NTN protein was constructed. NTN-c17.2, Mock-c17.2 and PBS were respectively transplanted in the striatum of rats in each group. NTN-c17.2, Mock-c17.2 and PBS Six days later 6-hydroxydopamine (6-OHDA) was injected. The differentiation of c17.2 neural stem cells, the expression of NTN gene and the protective effect of NTN protein were observed by immunohistochemistry, in situ hybridization, rotation behavior and neurotransmitter changes. Results After NTN-c17.2 cells were transplanted, the cells differentiated but still continued to secrete NTN protein. NTN protein is transported reversibly to the substantia nigra and protects tyrosine hydroxylase (TH) -positive neurons from damage by 6-OHDA. The dynamic observation of the rotating behavior of the animal model showed that the protective effect of NTN lasted for at least 4 months, and the NTN group was significantly better than the Mock and PBS groups. At 10 months after transplantation, the neurotransmitters such as dopamine concentration in the NTN and Mock groups were 95% (P <0.05) and 93% (P <0.05), respectively Group is 75%. Conclusion Transplantation of neural stem cells with NTN gene for Parkinson’s disease shows good protective effect.