神经元损伤修复机制研究的离体模型--体外培养原代神经元损伤模型的建立及损伤后c-Fos蛋白表达

目的：建立一个模拟脊髓全横断损伤后脊髓组分——原代培养的脊髓神经元损伤的模型。方法：取Wistar大鼠(孕14d)脊髓，培养10～12d后，采用所设计的标准模板，直视下进行损伤。观察损伤前后不同时间神经元的形态及即刻早期基因c-fos的表达变化。结果：在损伤8～12h后，损伤两侧存活的神经元突起便开始朝着损伤部位生长，随着时间的延长，神经元突起甚至可跨越损伤处而延伸至对侧。c-fos基因在损伤后10min即有表达，2h达到最高峰，随后逐渐下降。结论：此模型可用于模拟体外培养的脊髓神经元受到机械损伤以及观察神经元损伤后进行修复的一些分子事件，简便而可靠，实用性较强。

Establishment of a model for exploring molecular mechanism in repair of neuron injury:a method of mechanically injuring in vitro cultured neurons and expression of c-Fos protein after injury

Objective:To establish a model to investigate the mechanism of injury and regeneration of primary neuron, the most important element in spinal cord.Methods:Spinal cord of fetal Wistar rats (14 th gestation day) was taken and its neurons were cultured for 10-12 days, then injury was made on the culture disk with a self_made standard template under direct vision. Changes in neuronal morphology and expression of c_fos gene, one of the immediate_early genes, were observed before and after injury at different intervals.Results:The remaining proximal processes of survived neurons in both sides of the injury track began to grow toward the track 8-12 hours after injury, and they were even across the track and reached the opposite sides thereafter. The expression product of c_fos gene was detectable 10 min after injury and reached its peak at 2?hours, then decreased gradually.Conclusion:This model is easy and reliable for mechanically injuring in vitro cultured neurons from spinal cord and is suitable for observing molecular events involved in their repair process. Hence, it is of certain value in practical usage. [