半胱氨酸蛋白酶-3抑制剂对蛛网膜下腔出血后脑血管痉挛的作用

Effect of caspase-3 inhibitor on cerebral vasospasm after subarachnoid hemorrhage

目的观察早期使用大剂量特异性半胱氨酸蛋白酶-3(Caspase-3)抑制剂对蛛网膜下腔出血(SAH)后脑血管痉挛(CVS)的作用。方法将20只雄性新西兰白兔随机等分为4组:①对照组:枕大池注入生理盐水;②SAH组;③SAH+二甲基亚砜(DMSO)组;④SAH+Z-DEVD-FMK(特异性Caspase-3抑制剂)组。采用枕大池2次注血法建立兔SAH模型。2次注血后第5天采用灌流固定法处死动物,留取基底动脉标本。用免疫组化及原位细胞凋亡检测法(TUNEL)分别检测基底动脉内皮细胞Caspase-3的表达及凋亡情况,并通过测定基底动脉血管横截面积判断有无CVS。结果免疫组化显示SAH+Z-DEVD-FMK组基底动脉内皮细胞Caspase-3表达较SAH组及SAH+DMSO组显著性降低(P<0.05)。TUNEL染色示SAH+Z-DEVD-FMK组基底动脉内皮细胞凋亡程度较SAH组及SAH+DMSO组显著性减轻(P<0.01)。基底动脉横截面积测定结果提示SAH组及SAH+DMSO组较对照组显著性缩小(P<0.01),而SAH+Z-DEVD-FMK组较SAH组及SAH+DMSO组显著性增大(P<0.01)。结论早期使用大剂量Z-DEVD-FMK能减少兔SAH模型基底动脉内皮细胞的凋亡并减轻CVS。

Objective To observe the effect of early high-dose specific caspase-3 inhibitor on cerebral vasospasm （CVS） after subarachnoid hemorrhage （SAH）. Methods Twenty male New Zealand rabbits were equally randomized into 4 groups： ① control, injection of normal saline to the cistema magna,② SAH, ③ SAIl treated with DMSO, and ④ SAH treated with z-DEVD-fmk （a specific Caspase-3 inhibitor）, The rabbit SAH model was constructed by blood injection into the cistema magna twice. The perfusion-fixation was performed for sacrifice of the rabbits on the 5th day after the second blood-injection in vivo and the basilar artery was taken. Immunohistochemical technique and TUNEL were used to observe the expression of caspase-3 and apoptosis of endothelial cells in the basilar artery. The cross-sectional area of each basilar arterial lumen was measured to evaluate the degree of cerebral vasospasm. Results Immunohistochemical technique showed that the expression of endothelial caspase-3 of the basilar artery was significantly lower in group SAH treated with z-DEVD-fmk than in group SAH and group SAH treated with DMSO （P〈0.05）, and TUNEL revealed that apoptosis level of endothelial cells in group SAH treated with z：DEVD-fmk was significantly lower than in group SAH and group SAH treated with DMSO （P〈0.01）. The cross-sectional areas of the basilar arterial lumen in group SAH and group SAH treated with DMSO were smaller than that of control group （P〈0.01）, while those of group SAH treated with z-DEVD-fmk were larger than that of the group SAId and group SAH treated with DMSO （P〈0.01）. Conclusion Early high dose specific caspase-3 inhibitor can decrease endothelial apoptosis in the basilar artery in rabbit SAH model, and relieve CVS after SAH .