一氧化氮在缺氧复氧性神经元凋亡中的作用

目的观察神经元缺氧复氧损伤时NO的动态变化及其与神经元凋亡的关系。方法取孕13~15d ICR小鼠,无菌条件下对胎鼠大脑皮质神经元进行原代分离培养,建立缺氧复氧诱导的皮质神经元凋亡模型。用流式细胞仪检测细胞凋亡率,用硝酸还原酶法测定细胞培养上清液中NO的含量,用MTT测定细胞活性,并在电镜下观察细胞形态学变化。结果经缺氧复氧处理的神经元,随着缺氧时间延长细胞存活率逐渐下降,神经元凋亡率呈时间依赖性升高,NO含量逐渐升高,至缺氧8h复氧24h达到高峰,与正常对照组比较有显著性差异(P<0.01)。细胞超微结构呈现凋亡样改变。结论NO介导了缺氧复氧性神经元凋亡过程。

Effect of nitric oxide on cortical neuron apoptosis induced by anoxia-reoxygen

Objective To explore the relationship between cortical neuron apoptosis and the dynamic changes of nitric oxide(NO) during brain injury induced by anoxic - reoxygen. Methods Cortical neurons were removed from 13 ~ 15 - day - old embryonic ICR mice fetuses aseptically and primary cultivated in the L- 15 medium.The model of damage induced by anoxia - reoxygen was established on cultured cortical neurons.The neuron viability was measured by MTT assay, NO contents were determined by Nitrate reductase assay and the rate of neuronal apoptosis was measured by flow cytometer system.The changes of morphology were observed by electron microscopy. Results Compared to the control group, the cortical neuron survival decreased, the rate of neuron apoptosis step - up and the NO contents increased time - dependently in anoxia - reoxygen group and reach peak after H8R24( P < 0.01) .The most striking ultrastructural changes of apoptosis were observed by electron microscopy in anoxia - reoxygen neurons. Conclusion NO mediates neuron apoptosis during brain injury induced by anoxic - reoxygen.