摘要
目的:探讨神经生长因子(nervegrowthfactor,NGF)对小脑皮质神经元凋亡的影响,为临床治疗小脑变性疾病的提供新的措施。方法:实验于2001-12/2003-03在锦州医学院科技楼实验室完成。以原代培养的新生SD乳鼠小脑皮质细胞建立谷氨酸诱导的神经细胞凋亡模型。通过流式细胞术检测神经细胞凋亡率;利用光学显微镜技术观察细胞凋亡的形态学改变。结果:谷氨酸(500μmol/L)作用5min可诱导小脑神经细胞凋亡。应用流式细胞仪检测发现受损细胞在G1峰前出现一个G1亚峰,凋亡细胞所占比例为50.2%及45.7%,通过荧光染色观察发现模型组细胞核出现染色体聚集,凋亡小体形成等形态学变化。流式细胞仪检测结果显示模型组与正常对照组亦有明显差异,光学和荧光显微镜观察发现受损细胞的形态学变化也得到明显改善。结论:外源性NGF能够减轻大鼠小脑皮质神经细胞凋亡。
AIM: To study the effect of nerve growth factors(NGF) on neural apoptosis in cerebellar cortex of rats so as to provide new therapeutic methods to treat cerebellar degenerated diseases.METHODS: The experiment was finished in Jinzhou Medical College from December 2001 to March 2003.Neuronal apoptosis models induced by aminoglutaric acid were established by using the cells in cerebellar cortex of primary cultured neonatal SD rats. The apoptosis was detected by using flow cytometry (FCM); morphological changes of apoptosis were estimated with light microscope and fluorescence microscope.RESULTS: Injection with glutamic acid(500 μ mol/L) for 5 minutes induced the apoptosis of cultured neurons from the cerebella of SD rats.The results of FCM showed a sub- G1 spike indicated before G1 spike. The rate of neuronal apoptosis was 50.24% and 45.7% , respectively. The aggregation of chromosome and apoptosis body were observed under fluorescent microscope .The results of MTT showed that survival rate of cells of NGF in high and moderate dosage groups was significantly higher than that in model group, the content of LDH was significantly reduced and the concentration SDH was significantly increased. The results of FCM showed that there was significant difference between model group and control group. By the use of light microscope and fluorescence microscope, morphological changes of damaged cells were improved.CONCLUSION:Exogenous NGF can relieve the apoptosis of neurons in cerebellar cortex of rats.
出处
《中国临床康复》
CSCD
北大核心
2005年第9期50-51,i002,共3页
Chinese Journal of Clinical Rehabilitation
基金
辽宁省教育厅重点科研课题(991721597)~~
