体外培养原代神经元损伤模型的建立及损伤后脑红蛋白的表达

Establishment of a mechanical injury model of primary cultured neurons in vitro and expression of neuroglobin after injury

目的建立原代神经元机械性损伤的体外细胞模型并探讨神经元损伤后脑红蛋白(NGB)的表达变化。方法取出生12h内Wistar鼠的大脑皮质,培养10￣12d后,采用所设计的标准模板,直视下进行损伤,采用免疫组化方法和图像分析技术观测损伤前、后不同时间点神经元的NGB免疫组化反应及其阳性信号的灰度值。结果皮质神经元损伤2h后,NGB阳性信号强度即开始增加,16h后达高峰,随后逐渐下降,128h后恢复至损伤前水平。结论皮质神经元体外损伤模型可用于在体外观察神经元受到机械性损伤后细胞内蛋白表达变化的一些分子事件,简单实用。机械性损伤可以诱导神经元NGB在损伤后一定时间段内表达上调。

Objective To establish a mechanical injury model of primary cultured cerebral neurons in vitro, and explore the expression of neuroglobin in the injured neuron. Methods Brain of Wistar rat （within 12 h post birth） was taken and the neurons were cultured for 10-12 days, then injury was made on a self-made standard template under direct vision. The immunohistochemical response of neuroglobin in the neurons and gray values of immunoreactive products were studied by immunohistochemical method and image analysis before and after injury at different intervals. Results The expression of neuroglobin began to increase at 2 h after injury and reached its peak at 16 h, then decreased gradually and recovered to the level before injury at 128 h. Conclusion This model is easy and reliable for mechanical injury of cultured neurons in vitro from brain and suitable for observing molecular events such as expression changes of a certain protein. And mechanical injury can induce the expression up-regulation of NGB in neuron injury in a certain time.