【摘要】 目的　构建一种可分泌人胶质细胞源性神经营养因子 (GDNF)的基因工程细胞 ,并探讨其在基因治疗帕金森病中的可能作用。方法　克隆含Kozak序列的人GDNFcDNA ,并将其转染至人类神经母细胞瘤细胞系SH SY5Y细胞 ,用RT PCR方法筛选阳性细胞克隆 ,将此工程细胞与大鼠原代多巴胺能神经元共培养 ,免疫组织化学检测多巴胺能神经元。结果　 (1)基因工程细胞可防止多巴胺能神经元退变死亡 ,与对照组比较 ,最多增加了 95 4 %,P <0 0 1。 (2 )工程细胞可抵抗MPP+ 对多巴胺能神经元的毒性损伤作用 ,与对照组相比 ,细胞数目增加了 9 5～ 10 8倍 ,P <0 0 1。结论　首次构建了可分泌人GDNF的工程细胞 ,其对多巴胺能神经元具有明显的营养保护作用 ,其在帕金森病的基因治疗中可能具有重要的应用价值。更多还原

【Abstract】 Objective To construct a kind of engineered cell secreting human GDNF and study its possible effects on gene therapy of Parkinson′s disease. Method Human GDNF cDNA with Kozak sequence was cloned by RT-PCR, and then was transfected into SH-SY5Y cell line of human neuroblastoma. These engineered cells were co-cultured with primary mesencephalic cells of rats. Dopaminergic neurons were examined by immunohistochemistry. Results The number of dopaminergic neurons protected by engineered cells increased at least by 95.4% in comparison with the control cells (P<0.01). The number of dopaminergic neurons protected by engineered cells against MPP + toxicity increased 9.5～10.8 times (P<0.01). Conclusion A kind of engineered SH-SY5Y cells secreting human GDNF has been constructed successfully. These cells obviously protect dopaminergic neurons against degeneration and MPP + toxication and may play an important role in gene therapy of Parkinson′s disease.更多还原