摘要
目的揭示人脑急性创伤性脑损伤后Omi/Htr A2介导的神经细胞凋亡线粒体途径的激活。方法选择100例脑外伤开颅手术中获得的脑挫裂伤脑组织作为外伤组,另外选择100例脑出血患者手术径路中获得的正常脑组织作为对照组,采用流式细胞仪检测神经细胞凋亡,采用Western-blot检测Omi/Htr A2、X染色体连锁凋亡抑制蛋白(XIAP)、pro-caspase 3、pro-caspase 9和剪切聚腺苷二磷酸-核糖多聚酶(PARP)表达,采用四肽荧光底物法检测caspase 3和caspase 9活性。结果外伤组脑组织凋亡神经细胞比例、Omi/Htr A2、pro-caspase 3、pro-caspase 9和剪切PARP表达及caspase 3和caspase 9活性较对照组显著升高(均P<0.01),外伤组脑组织XIAP表达较对照组显著下降(P<0.01)。外伤组脑组织Omi/Htr A2表达与凋亡神经细胞比率、pro-caspase 3、pro-caspase 9和剪切PARP表达及caspase 3和caspase 9活性呈显著正相关(均P<0.01),与XIAP表达呈显著负相关(P<0.01);外伤组脑组织XIAP表达与凋亡神经细胞比率、pro-caspase 3、pro-caspase 9和剪切PARP表达及caspase 3和caspase 9活性呈显著负相关(均P<0.01)。结论 Omi/Htr A2介导的线粒体途径可能参与人急性创伤性脑损伤后神经细胞凋亡过程。
Objective To investigate the effect of Omi/Htr A2 mediated mitochondrial signaling pathway in neuronal apoptosis after acute traumatic brain injury. Methods Brain tissue samples were collected during craniotomy from 100 patients with cerebral contusion and laceration(trauma group), and normal brain tissue samples were collected from 100 patients with intracerebral hemorrhage(control group). Flow cytometry was applied to determine apoptotic neuronal cells, Western-blotting was performed to test expressions of Omi/Htr A2, X-linked inhibitor of apoptosis protein(XIAP), pro-caspase-3, pro-caspase-9 and cleaved poly ADP-ribose polymerase(PARP), and four peptide fluorescence substrate method was employed to detect activities of caspase 3 and caspase 9 proteins. Results Percentage of apoptotic nerve cells, expressions of Omi/Htr A2, pro-caspase 3,pro-caspase 9 and cleaved PARP as well as activities of caspase 3 and caspase 9 proteins were significantly higher and the expression of XIAP was significantly lower in brain tissues of trauma group than those in control group(all P <0.01). In trauma group, expression of Omi/Htr A2 was positively correlated with percentage of apoptotic nerve cells, expressions of pro-caspase 3,pro-caspase 9 and cleaved PARP as well as activities of caspase 3 and caspase 9 proteins, and negatively correlation with the expression of XIAP. The expression of XIAP was negatively correlated with percentage of apoptotic nerve cells, expressions of Omi/Htr A2, pro-caspase 3, pro-caspase 9 and cleaved PARP as well as activities of caspase 3 and caspase 9 proteins.Conclusion Omi/Htr A2 mediated mitochondrial signaling pathway may be involved in neuronal apoptosis after acute traumatic brain injury.
引文
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