摘要
目的 :观察MN9D细胞中Nurr1的表达和分布及Nurr1过表达对酪氨酸羟化酶的影响。 方法 :应用高压液相色谱、免疫细胞化学、转基因、Westernblot和Northernblot等方法检测了MN9D细胞中多巴胺及其代谢物的水平和Nurr1蛋白在细胞中的分布 ,建立了过表达Nurr1的细胞株并观察了Nurr1过表达对TH基因的影响。结果 :MN9D细胞裂解液中含有大量多巴胺及其代谢物 ;Nurr1蛋白主要分布在MN9D细胞的胞质中 ,尤以核周最多 ;MN9D细胞中有Nurr1的mRNA存在和蛋白质的表达 ;过表达Nurr1的MN9D细胞A1株酪氨酸羟化酶表达增高。结论 :Nurr1与多巴胺能神经元发育有关 ,有可能用于帕金森病的基因治疗。
Objective: To study the expression and localization of Nurr1 in MN 9D cells and the influence of Nurr1 overexpression on the expression of tyrosine hydroxylase (TH) in vitro . Methods: Dopamine and its metabolites,DOPAC and HVA,were detected in MN9D cells with HPLC. The mRNA and protein of Nurr1 were detected with Northern blot and Western blot. Results: MN9D expressed Nurr1 mainly in the cytoplasm. The expression of TH was increased in an A1 cell line derived from the Nurr1 overexpression MN 9D cells. Conclusion:Nurr1 overexpression in the MN9D cells could promote the expression of TH in the cells, suggesting that the gene of Nurr1 was essential for development of dopaminergic neurons and might be potentially used for gene therapy of Parkinson's disease.
基金
国家重点基础研究计划 (G19990 5 40 0 8)
国家自然科学基金 ( 3 0 2 70 43 3 )
