胞二磷胆碱对大鼠弥漫性轴索损伤后神经元保护作用的研究

The effect of cytidine diphosphate choline on neuroprotection following diffuse axonal injury in rats

目的探讨胞二磷胆碱对大鼠弥漫性轴索损伤(diffuse axonal injury,DAI)后脑细胞功能变化的影响,评估药物的治疗价值。方法采用大鼠头部瞬间旋转损伤装置制备DAI模型,通过胞二磷胆碱及神经元凋亡通路抑制剂辛伐他汀干预控制,对比观察胞二磷胆碱对DAI大鼠神经修复的作用,于术后24、48、72 h及7d提取脑组织标本,western blot法检测SDF-1和神经元凋亡及死亡相关蛋白激酶1(DAPK1)的表达变化,并检测β-APP的表达来指示轴突的损伤程度,用免疫荧光染色指示Rh OA和NOGO-A表达的相关性。结果 DAI模型组24 h后均出现明显的神经细胞坏死和轴突变性等病理改变;大脑皮层Rh OA、NOGO-A、SDF-1和DAPK1随着DAI后时间的延长而增加,经免疫荧光双染色显示两者在脑内的表达有显著的同一性;胞二磷胆碱及辛伐他汀能明显降低SDF-1和DAPK1的表达,胞二磷胆碱作用更加显著。结论胞二磷胆碱可明显改善DAI后神经功能并减轻轴突损伤。

Objective To investigate the effect of cytidine diphosphate cholineon neuroprotection following diffuse axonal injuryso as to evaluate the therapeutic effect. Methods The DAI model was prepared by using a coronal rotation device,and the brain tissue was obtained at 24 h,48 h and 72 h after DAI. The effect of neuroprotection of cytidine diphosphate and neuronal apoptosissignaling pathway inhibitor simvastatin was observed contrastively,The expression of SDF-1 and DAPK1 was detected by western blot,and the expression of β-APP as an indicater for the severity of axonal injury. The immunofluorescence double staining was used to observe the correlation of NOGO-A and Rh OA expression.Results There were pathologic changes in the rats at 24 h after DAI such as neuronal death and axonal disruption.The expression of Rh OA,SDF-1,NOGO-A and DAPK1 were in the consistent trend in that all of them increased with prolonged time duration and were significantly increased in cerebral cortex at 24 h,48 h,72 h and 7d after DAI compared with that in the control group.The expression of β-APP was also significantly increased at 24 h,48 h,72 h and 7d after DAI compared with that in the control group. On the contrary,after the administration of cytidine diphosphate choline and simvastatin,the level of Rh OA,SDF-1,NOGO-A and DAPK1 were also markedly decreased in the cytidine diphosphate choline and simvastatin group compared with those in DAI group,further more the cytidine diphosphate cholinegroup has a more obvious suppression.Conclusion The cytidine diphosphate choline can significantly improve the neurological function and repress the axonaldisruptionafter DAI.