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目的观察早期使用大剂量特异性半胱氨酸蛋白酶-3(caspase-3)抑制剂对兔蛛网膜下腔出血(SAH)后脑皮层神经元的凋亡的影响。方法枕大池二次注血法建立兔SAH模型。一次注血后第5天灌流固定法处死动物。用免疫组化及凋亡细胞原位末端标记技术(TUNEL)分别检测颞叶皮层神经元caspase-3的表达及凋亡情况。结果免疫组化显示SAH+Z-DEVD-FMK(一种特异性caspase-3抑制剂)组皮层神经元caspase-3表达较SAH组及SAH+二甲基亚砜(DMSO,有机溶剂)组明显降低(P＜0．05)；TUNEL染色结合凋亡指数(AI)显示SAH组及SAH+DMSO组较对照组存在明显皮层神经元凋亡(P＜0．01),而SAH+Z-DEVD-FMK组皮层神经元凋亡较SAH组及SAH+DMSO组明显减轻(P＜0．01)。结论早期使用特异性大剂量caspase-3抑制剂能减轻兔SAH后脑皮层神经元的凋亡,起到脑保护的作用。
Objective To observe the effect of early high-dose caspase-3 inhibitor on the apoptosis of cortical neurons in rabbits after subarachnoid hemorrhage (SAH). Methods The rabbit model of SAH was established by the secondary injection of occipital cistern. Animals were sacrificed by perfusion fixation on day 5 after a blood injection. The expression and apoptosis of caspase-3 in temporal cortex neurons were detected by immunohistochemistry and TUNEL. Results Immunohistochemistry showed that the expression of caspase-3 in cortical neurons of SAH + Z-DEVD-FMK (a specific caspase-3 inhibitor) group was significantly lower than that of SAH group and SAH + dimethyl sulfoxide (DMSO, organic solvent) (P <0.05). The apoptosis of SAH group and SAH + DMSO group was significantly higher than that of the control group (P <0.01) by TUNEL staining and apoptosis index (AI) Compared with SAH group and SAH + DMSO group, the apoptosis of cortical neurons in FMK group was significantly reduced (P <0.01). Conclusion Early use of specific high-dose caspase-3 inhibitors can reduce apoptosis of cerebral cortex neurons in rabbits after SAH, play a role in brain protection.