移植的神经干细胞在帕金森病模型小鼠脑内的存活与分化

Survival and differentiation of transplanted neural stem cells in the brain of MPTP-induced Parkinson's disease model mice

目的观察移植的神经干细胞在帕金森病 (PD)模型小鼠脑内的存活与分化。方法C5 7BL 6小鼠皮下注射 1 甲基 4 苯基 1,2 ,3 ,6 四氢吡啶 (MPTP)造成PD模型。用 5 溴尿嘧啶 (BrdU)标记的神经干细胞分别移植到模型鼠的一侧或两侧纹状体。用酪氨酸羟化酶免疫荧光评价MPTP对黑质多巴胺能神经元的毒性。用免疫组织化学和激光共聚焦鉴定移植的神经干细胞在体内的存活与分化。结果MPTP使黑质酪氨酸羟化酶阳性细胞显著减少 ;在细胞移植的纹状体内发现有明显散在分布的BrdU阳性细胞 ,表明移植的神经干细胞已经存活 ;激光共聚焦显示部分BrdU阳性细胞已分化为酪氨酸羟化酶阳性细胞。结论移植的神经干细胞能在PD模型小鼠纹状体存活 ,并可分化出特定的多巴胺能神经元。

ObjectiveTo determine survival and differentiation of cultured neural stem cells (NSCs) into viable and functional neurons upon transplantation into the brain of MPTP-induced Parkinson's Disease (PD) mode micel.MethodsMice model of PD was established with intraperitoneal (i.p.) injections of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP,40mg/kg) twice interspersed by 16 hr. Tissues isolated from an embryonic rat midbrain were cultured in clonal density. After labeled with 5-bromo-2′-deoxyuridine (BrdU), the NSCs were transplanted into the uni- or bilateral striatum of PD mice. Tyrosine hydroxylase (TH) immunofluorescence was used to evaluate the toxicity of MPTP on the neural cells in the substantia nigra. Immunohistochemistry and laser confocal were used to detect the survival and differentiation of transplanted NSCs.ResultsTH positive neural cells were significantly reduced in the substantia nigra after MPTP injection. Immunohistology showed that the uni- or bilateral transplanted NSCs could survive in the brains of PD model mice.Some transplanted NSCs could properly differentiate into targeted TH positive neural cells in vivo.ConclusionsThe transplanted multipotent NSCs could survive, differentiate into functional dopamine neurons in the brains of PD model mice.