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目的观察人脑急性创伤性脑损伤后神经细胞凋亡及其与caspase-3基因表达的关系。方法术中采集12例重型颅脑损伤患者挫裂伤脑组织和脑叶切除组织标本15个,分别应用凋亡细胞原位末端标记法(TUNEL)、DNA琼脂糖凝胶电泳、透射电镜等检测神经细胞凋亡的变化,采用免疫组织化学技术检测caspase-3蛋白的表达,并借助caspase-3荧光分析试剂盒检测caspase-3蛋白酶活性的变化。结果12个挫裂伤脑组织标本中10个透射电镜发现有散在分布、数量不等的凋亡神经细胞；TUNEL染色11个出现阳性凋亡神经细胞；5个DNA出现凋亡特异性的“梯状”电泳带；9个出现caspase-3蛋白超表达；11个caspase-3蛋白酶活性明显增高；凋亡出现时间从伤后4h直到216h,高峰在23～72h。结论急性创伤性脑损伤患者存在神经细胞凋亡,并且具有时间和空问依赖性。Caspase-3的超表达与激活参与了人脑创伤性脑损伤后的细胞凋亡过程。
Objective To observe the relationship between neuronal apoptosis and caspase-3 gene expression after acute traumatic brain injury in human brain. Methods Fifteen patients with contusion and laceration were collected from 12 patients with severe craniocerebral injury during operation. TUNEL, DNA agarose gel electrophoresis and transmission electron microscopy The changes of caspase-3 protein expression were detected by immunohistochemistry and the changes of caspase-3 protease activity were detected by caspase-3 fluorescence analysis kit. Results Ten apoptotic brain tissue samples were found by transmission electron microscopy. A total of 11 neuronal apoptotic neurons were found in TUNEL staining, and 5 apoptotic cells 9 "caspase-3 protein overexpression; 11 caspase-3 protease activity was significantly increased; apoptosis occurred from 4h until 216h after injury, peaked at 23 ~ 72h. Conclusion Apoptosis of neurons exists in patients with acute traumatic brain injury and is dependent on time and space. Overexpression and activation of Caspase-3 are involved in the process of apoptosis after traumatic brain injury in human brain.