摘要
目的 观察急性脑创伤后Caspase 3在神经元凋亡中的作用。方法 采用大鼠脑创伤模型 ,以TUNEL法观察皮层、海马区神经元凋亡 ;以Northern杂交、原位杂交、免疫组化、Western杂交等对Caspase 3在mRNA与蛋白质水平表达和酶活性变化进行观察 ;通过脑室注射给予Z DEVD .fmk ,观察其对神经元凋亡的治疗作用。结果 大鼠打击后 2小时可见TUNEL染色阳性神经元 ,1天最明显 ,7天时仍高。Northern杂交和原位杂交显示 ,皮层、海马区神经元致伤后 2小时Caspase 3mRNA表达增高 ,1天最明显 ,7天仍高于正常。免疫组化法见Caspase 3P2 0亚单位在打击后 2小时表达增高 ,1～ 3天最明显 ,可持续至 7天。Western杂交显示Caspase 3下游底物PARP被降解。脑室内注射Z DE VD .fmk可使神经元凋亡减少。结论 急性脑创伤后神经元中Caspase 3表达增高 ,酶活性增强 ,可导致神经元凋亡 ,Caspase 3特异性酶活性抑制剂能减少神经元凋亡。
Objective To analyze the role of Caspase 3 on neuronal apoptosis after acute brain trauma.Methods Experiments were based on rat brain injury model. After impact, neuronal apoptosis was observed by TUNEL stain.mRNA and protein expression and enzyme activation of Caspase 3 were observed by northern blot, in situ hybridization, immunohistochemistry stain and western blot.Z DEVD.fmk, a special Caspase 3 enzyme inhibitor was given to rat via intraventricular injection, and its therapeutic effect on neuronal apoptosis was observed.Results TUNEL stain show that neuronal apoptosis appeared 2 hours after injury, most peaked on 1 day,last to 7 day.Northern blot and in situ hybridization show that Caspase 3 mRNA expressed in cortex and hippocampal neurons, increased on 2 hours after injury, peaked on 1 day, which was 3 times higher than that of controls, last to 7 day.With the antibody for Caspase 3 P20 subunit, immunohistochemistry stain show that the active Caspase 3 expression increased 2 hours after injury, peaked on 1 3 days, continuously higher than that of controls to 7 days.The electrophoresis band of PARP degradation would be seen by western blot.Caspase 3 enzyme inhibitor could reduce neuronal apoptosis.Conclusions The increased expression of Caspase 3 mRNA and protein and enzyme activity in neurons after brain trauma are related to neuronal apoptosis, special Caspase 3 enzyme inhibitor can apparently improve the therapeutic effect on neuronal apoptosis.
出处
《中华神经外科杂志》
CSCD
北大核心
2002年第6期375-378,共4页
Chinese Journal of Neurosurgery
