GAD65基因转染神经干细胞的实验研究

目的检测神经干细胞经谷氨酸脱羧酶（GAD65）基因转染并诱导分化后γ-氨基丁酸（GABA）的表达，为GAD65基因修饰神经干细胞移植治疗癫痫提供理论依据。方法采用新一代高效脂质体作为转染试剂的基因转导方法转染神经干细胞，通过免疫细胞荧光方法鉴定神经干细胞分化为GABA阳性神经元情况。结果采用脂质体转染方法能成功地将GAD65基因转染至神经干细胞中，转染的神经干细胞在分化第7天及第14天GABA阳性神经元分别占（31．9±4．2）％、（26．5±2．3）％，未转染的神经干细胞分化的GABA阳性神经元则分别占（28.4±1．9）％和（20．3±3．5）％，转染者与未转染者相比，分化的GABA阳性神经元百分比有显著性差异（P〈0．05）。结论采用脂质体基因转染技术，可以将GAD65基因转染至神经干细胞中，并促进神经干细胞分化为GABA神经元。

Experimental Study of Expression of Gamma Aminobutyric Acid by Neural stem cells transfected with Glutamic Acid Decarboxylase Gene

Objectives To sbldy the expression of Gamma-aminobutyric acid (GABA) by neural stem cells (NSCs) transfected with dutamic acid decarboxylase (GAD65) gene in order to provide the basis for treatment of epilepsy by implanting NSCs transfected with GAD65 gene.Methods NSCs were transfected by PcDNA3-GAD65 liposomes which is a new generation of highly efficient transfection reagent.The GABA-positive neurons differentited by transfected NSCs with PcDNA3-GAD65 and non-transfected NSCs was identified by the method of immune cells fluorescent. Results NSCs were successfully transfected by PcDNA3-GAD65. The quantities of GABA-positive neutons were(31.9±4.2)% and (26.5±2.3)% respectively in the first 7 days and 14 days of dlfferentiation in the transfected group, and (28.4±1.9)% and (20.3±3.5)% respectively in the non-transfected group. There was a significant differenct in the quantity of GABA-positive neurons between both the transfected and non-transfected groups (P