目的 观察黄芪甲苷(AS-Ⅳ)对蛛网膜下腔出血(SAH)后迟发性脑血管痉挛(DCVS)的作用.方法 将雄性SD大鼠分为Sham组(A)、SAH+ AS-Ⅳ组(B)、SAH组(C)、SAH+vehicle组(D).采用枕大池二次注血法制作蛛网膜下腔出血后DCVS模型.造模后1～5 d,每天给予B组5％酒精助溶的AS-Ⅳ混悬液20 mg/kg;D组大鼠等体积的5％酒精;A、C组大鼠等体积的生理盐水.采用苏木素-伊红(HE)染色法观察脑基底动脉的形态学改变;原位缺口末端标记法(TUNEL)检测基底动脉内皮细胞和血管平滑肌细胞凋亡;免疫组织化学染色测定基底动脉上裂解的半胱氨酰天冬氨酸特异性蛋白酶-3(cleaved Caspase-3)的表达;采用Western blot检测颅底血管B细胞淋巴瘤/白血病-2(bcl-2)、bcl-2相关X蛋白(bax)、cbaved Caspase-3的表达.结果 与Sham组大鼠比较,B、C、D组的基底动脉管壁增厚,管腔横截面积变小;基底动脉上bcl-2表达减少(P＜0.05),bax和Cleaved Caspase-3的表达增加(P＜0.05).给予AS-Ⅳ干预后,与SAH+ vehicle、SAH组比较,SAH+ AS-Ⅳ组中大鼠的bel-2表达增加(P＜0.05),bax和cleaved Caspase-3的表达减少(CleavedCaspase-3:0.030 8±0.003 2比0.0704±0.0060、0.0690±0.005 7,P＜0.05).结论 AS-Ⅳ可缓解大鼠SAH后DCVS,其机制可能与bcl-2、bax、Caspase-3介导的凋亡信号通路有关.%Objective Investigate the effect of Astragaloside Ⅳ (AS-Ⅳ) on delayed cerebral vasospasm (DCVS) following subarachnoid hemorrhage.Methods The male SD rats were divided into Sham group (A),SAH +AS-Ⅳ group (B),SAH model group (C) and SAH +vehicle group (D).The model of DCVS following subarachnoid hemorrhage was made by the double injection method of occipital cistern.During the 5 days after model success,rats in the B group were treated with daily AS-Ⅳ (20 mg/kg) which was dissolved in 5％ alcohol;rats in the D group were injected with identical volume of physiological saline containing 5％ alcohol;rats in the SAH group and the Sham group were inject with identical volume of physiological saline.Morphological changes of basilar artery was observed by hematoxylin and eosin (HE) staining.The apoptosis of basilar artery endothelial cells and vascular smooth muscle cells was detected by terminal-deoxynucleotidyl transferase mediated nick end labeling (TUNEL) staining.The cleaved cysteinyl aspartate-specific protease (Caspase)-3 expression in basilar artery was detected by immunohistochemical staining.The expression of B cell lymphoma/leukemia-2 (bcl-2),bcl-2 associated X protein (bax) and cleaved Caspase-3 was detected by Western blotting.Results Compared with Sham group,the basilar artery wall became thicker and lumen cross-sectional area became smaller.The expression of bax and cleaved Caspase-3 (P ＜ 0.05) was decreased in basilar artery (P ＜0.05).Compared with SAH + vehicle group and SAH group,the expression of cleaved Caspase-3 in SAH + AS-Ⅳ group increased (P ＜0.05) and the expression of bax and cleaved Caspase-3 decreased after AS-Ⅳ intervention (Cleaved Caspase-3:0.030 8 ± 0.003 2 vs.0.070 4 ± 0.006 0,0.069 0 ±0.005 7,P ＜ 0.05).Conclusion AS-Ⅳ can relieve delayed cerebral vasospasm and its mechanism may be related to the fact that AS-Ⅳ can interfere with the apoptosis signaling pathway mediated by bcl-2,bax and Caspase-3.
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