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目的:探讨体外培养大鼠海马神经元癫模型中神经元丢失的分子机制。方法:制备大鼠海马神经元癫样放电细胞模型,以膜片钳全细胞记录对模型放电进行检测,采用RT PCR法克隆大鼠全长caspase 3 cDNA,采用原位杂交和流式细胞术检测模型中caspase 3基因表达和神经元凋亡情况。结果:成功克隆获得大鼠全长caspase 3 cDNA,其序列与文献报道一致。模型组海马神经元癫样放电3 h后caspase 3基因表达开始增多,6 h后凋亡细胞开始明显增加,两者均较对照组明显增加。结论:癫样放电可能启动神经元caspase 3表达,继而介导神经元凋亡。
Objective: To investigate the molecular mechanism of neuron loss in epileptic model of rat hippocampal neurons cultured in vitro. Methods: The epileptiform discharge cell model of rat hippocampal neurons was prepared. Whole-cell patch clamp recording was used to detect the model discharge. The full-length rat caspase 3 cDNA was cloned by RT-PCR. The in situ hybridization and flow cytometry Detection of caspase 3 gene expression and neuronal apoptosis in the model. Results: The full-length rat caspase 3 cDNA was successfully cloned and its sequence was consistent with that reported in the literature. The expression of caspase 3 began to increase 3 hours after epileptiform discharge in hippocampal neurons of model group, and the apoptotic cells began to increase significantly after 6 hours, both of which increased significantly compared with the control group. CONCLUSION: Epileptiform discharges may activate caspase 3 expression in neurons and subsequently induce neuronal apoptosis.