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目的探讨内皮细胞凋亡在兔蛛网膜下腔出血(SAH)后脑血管痉挛形成中的作用。方法实验分正常组、对照组(枕大池注入生理盐水)、SAH3d组、SAH5d组、SAH7d组和SAH10d组。采用自体动脉血枕大池注入方法建立SAH模型。应用脑血管造影观察基底动脉形态改变,透射电镜和TUNEL技术观察内皮细胞凋亡,免疫组化法检测Caspase-3蛋白表达。结果脑血管造影发现,SAH后第3天基底动脉狭窄,第7天达高峰,第10天缓解。SAH5d组和SAH7d组基底动脉壁出现胞浆浓缩、核染色质浓缩边集等凋亡样改变的内皮细胞和较多TUNEL染色阳性内皮细胞,同时内皮细胞中Caspase-3蛋白表达上调,SAH7d组尤为明显;SAH3d组和SAH10d组内皮细胞凋亡样病理改变减轻,TUNEL染色阳性细胞明显减少,内皮细胞Caspase-3蛋白表达减弱;正常组和对照组未见明显病理改变。结论脑血管壁存在的内皮细胞凋亡可能在SAH后脑血管痉挛形成中起重要作用。
Objective To investigate the role of endothelial cell apoptosis in the formation of cerebral vasospasm after subarachnoid hemorrhage (SAH) in rabbits. Methods The experiment was divided into normal group, control group (physiological saline injected into occipital cistern), SAH3d group, SAH5d group, SAH7d group and SAH10d group. The model of SAH was established by injecting autologous arterial blood-fat pool. Cerebral angiography was used to observe the morphological changes of basilar artery. The apoptosis of endothelial cells was observed by transmission electron microscopy and TUNEL technique. Caspase-3 protein expression was detected by immunohistochemistry. Results Cerebral angiography showed basilar artery stenosis on the third day after SAH, reaching the peak on the seventh day and remission on the tenth day. In the SAH5d group and the SAH7d group, apoptotic-like endothelial cells and more TUNEL-stained endothelial cells appeared in the basilar artery wall of the SAH5d group, and the expression of Caspase-3 protein was up-regulated in the endothelial cells, especially in the SAH7d group Obviously, the apoptotic pathological changes of endothelial cells in SAH3d group and SAH10d group were alleviated, the number of TUNEL-positive cells and the expression of Caspase-3 in endothelial cells were decreased. No obvious pathological changes were observed in normal group and control group. Conclusion The apoptosis of endothelial cells in cerebrovascular wall may play an important role in the formation of cerebral vasospasm after SAH.