NGF对新生大鼠小脑皮质神经元凋亡的影响

Effect of NGF on Neuronal Apoptosis in Cerebellar Cortex of Cultured Rats

目的 研究NGF对新生大鼠小脑皮质神经细胞凋亡的影响。为其临床治疗小脑变性疾病奠定理论基础。方法 用原代培养的大鼠新生乳鼠小脑皮质细胞建立谷氨酸诱导的神经细胞凋亡模型 ,实验组的各组提前 2 4h加入NGF ,模型组不给NGF ,并观察NGF的保护作用。通过检测培养液LDH含量 ,反映细胞的代谢情况和细胞功能和状态 ,通过流式细胞仪检测DNA的含量 ,定性的反映细胞凋亡率。结果 谷氨酸 (Glu) 5 0 0 μmol·L-1与细胞作用时间 5min可诱导细胞凋亡。取细胞培养液检测LDH的浓度明显增高 ,流式细胞仪检测显示 ,在G1峰前出现一个“G1亚峰”凋亡细胞 ,所占比例为 5 0 2 %及 4 5 7%。NGF (5 0 ,10 0 μg·L-1)能够使LDH (乳酸脱氢酶 )的含量减低 ,凋亡细胞数减少。

Objective To investigate the effect of NGF on neuronal apoptosis in cultured rat cerebellar neurons. Methods The neuro protective effects of NGF on neuronal apoptosis induced by glutamate were studied in rat primary cerebellum cortical cultures.In treatment groups,NGF was treated at 24h before Glu was added.The content of LDH in the cell supermate and neuronal viability were measured.The rates of neuronal apoptosis were measured through FCM. Results The neuronal apoptosis was induced by glutamate (Glu,500μmol/L)in cultured rat cerebellar neurons.In conrtol group,the LDH activity in the cell supermate significantly increased.FCM was used to detect the rate of neuronal apoptosis.There is a 'Sub-G 1'spike before G 1 spike.The percentage rate of neuronal apoptosis was 50 24% and 45 7% respectively.The aggregation of chromosome and apoptosis body was observed through fluorescent microscope.NGF(50,100μg/L)reduced the number of neuronal apoptosis,and the level of LDH.It can also relieve neuronal morphologic abnormality. Conclusions The results suggest that NGF can relieve neuronal apoptosis induced by EAA.