人GDNFcDNA在SH-SY5Y细胞中的表达及其对多巴胺能神经元的保护作用探讨

为建立一种可分泌人胶质细胞源性神经营养因子（GDNF）的工程细胞,研究其在帕金森病基因治疗中的可能作用。克隆携带Kozak序列的人GDNFcDNA,并转染至人类神经母细胞瘤细胞系SH－SY5Y细胞,将此工程细胞与大鼠原代多巴胺能神经元共培养,免疫组化检测多巴胺能神经元。发现工程细胞可防止多巴胺能神经元退变死亡,有助于多巴胺能神经元抵抗MPP^ 的毒性损伤。表明可分泌人GDNF的工程细胞在帕金森病基因治疗中可能具有重要的应用价值。

Expressing and ability protecting primary dopaminergic nerous of human GDNFcDNA in engineeredSH-SY5Y cells

To construct a kind of engineered cells that can secrete human glial cell line-derived neurotrophic factor（GDNF）and study its possible effects on gene therapy of Parkinson’s disease.Cloned the GDNFcDNA with Kozak sequence by RT-PCR,and then transfected it into SH-SY5Y cell line,co-cultured the engineered cells with primary mesencephalic cells.Results showed that the engineered cells showed obvious protecive effects such as anti-degeneration and anti-MPP＋lesion.GDNF produced by the engineered cells was of full biological function.It suggests that the engineered SH-SY5Y cells can secrete human GDNF and could play an important role in gene therapy of Parkinson’s disease.