心肌营养素-1对抗NMDA受体脑炎致海马神经元损伤的保护作用

Protective Effects of Cardiotrophin-1 on Hippocampal Neurons Injured by Anti-NMDA-receptor Encephalitis

目的:研究营养素-1(CT-1)干预治疗抗N-甲基-D-天门冬氨酸(NMDA)受体脑炎诱导的神经元损伤的保护作用及机制,为抗NMDA受体脑炎提供新的治疗靶点。方法:选择刚出生的SD大鼠培养原代神经元细胞,把原代神经元细胞分为实验组﹑对照组和正常组,分别建立抗NMDA受体脑炎诱导的神经元损伤模型,在实验组中加入CT-1(10 ng/mL),应用台盼蓝染色法、免疫组织化学和TUNEL细胞凋亡检测等技术,检测CT-1对抗NMDA受体脑炎诱导神经元损伤的保护作用以及细胞凋亡基因Caspase-3表达水平。结果:各种浓度的谷氨酸均能诱导神经元凋亡,谷氨酸浓度为100﹑200μmol/L时,细胞凋亡率高而死亡率并不高;加入CT-1神经元细胞,第1、2、3天,实验组细胞存活率均高于对照组,细胞凋亡率、Caspase-3阳性细胞率均低于对照组,差异均有统计学意义(P<0.05)。结论:CT-1对抗NMDA受体脑炎导致的神经元损伤有保护作用,可能是通过促使凋亡基因Caspase-3表达下调,减少细胞凋亡发生,从而对抗NMDA受体脑炎诱导的神经元损伤起到保护作用。

Objective:To study the protective effect and mechanism of cardiotrophin-1(CT-1)intervention on neuronal injury induced by anti-NMDA-receptor encephalitis and to provide a new therapeutic target for anti-NMDA-receptor encephalitis.Method:The primary neuron cells were cultured in newly born SD rats,they were divided into experimental group,control group and normal group,neuronal injury models induced by anti-NMDA-receptor encephalitis were established,CT-1(10 ng/mL)was added to experimental group.Trypan blue staining,immunohistochemistry and TUNEL cell apoptosis detection were used to detect the protective effect of CT-1 on neuronal injury induced by NMDA receptor encephalitis and the expression level of Caspase-3.Result:All concentrations of glutamate could induce neuronal apoptosis,when the concentration of glutamate was 100 and 200μmol/L,the apoptotic rate was high and the mortality rate was not high.On the 1st,2nd and 3rd day after adding CT-l neurons,the survival rate of experimental group were higher than those of control group,and the apoptotic rate and Caspase-3 positive cell rate were lower than those of control group,the differences were statistically significant(P<0.05).Conclusion:CT-1 has protective effect against neuronal injury induced by anti-NMDA-receptor encephalitis,which may be through down-regulation of Caspase-3 expression and reduction of apoptosis,thereby protecting neurons from NMDA receptor encephalitis-induced neuronal injury.