藁本内酯对大鼠蛛网膜下腔出血后迟发性血管痉挛的影响  

The effects of Z-Ligustilide on delayed cerebral vasospasm after subarachnoid hemorrhage in rats

作　　者：汪小青 竹梅 李英博 陈笛 Wang Xiaoqing;Zhu Mei;Li Yingbo;Chen Di(Department of Physiology,Faculty of Basic Medical Sciences,Chongqing Medical University,Chongqing 400010;Department of Physiology,Faculty of Basic Medical Sciences,Southwest Medical University,Luzhou 646000,China)

机构地区：[1]重庆医科大学基础医学院生理学教研室,重庆400010 [2]西南医科大学基础医学院生理学教研室,泸州646000

出　　处：《神经解剖学杂志》2020年第4期389-396,共8页Chinese Journal of Neuroanatomy

基　　金：重庆市科学技术委员会基础科学与前沿技术研究项目(cstc2017jcyjAX0358)。

摘　　要：目的:研究藁本内酯(LIG)对大鼠蛛网膜下腔出血(SAH)后迟发性脑血管痉挛(DCVS)的作用及其机制。方法:将100只SD大鼠采用随机数字表法分为:假手术组(Sham,n=22)、蛛网膜下腔出血组(SAH+vehicle,n=28)、藁本内酯5 mg组(SAH+LIG5,n=25)和藁本内酯20 mg组(SAH+LIG20,n=25)。采用改良的枕大池二次注血法制备大鼠SAH模型。第1次注血后30 min,SAH+LIG5组和SAH+LIG20组分别尾静脉注射藁本内酯5 mg/kg或20 mg/kg,SAH+vehicle组则注射等体积的溶剂(3%Tween 80),1次/d,连续7 d。根据Garcia神经功能评分法每天评定大鼠的神经功能。于第1次注血后7 d灌注取脑。尼氏染色观察皮层和海马区神经元形态改变;HE染色测量基底动脉横截面积及管壁厚度评估血管痉挛情况;TUNEL染色观察基底动脉内皮细胞的凋亡情况;Western Blot及免疫组化染色检测基底动脉p53、Bcl-2、Bax和cleaved caspase-3的表达情况。结果:LIG治疗可促进SAH大鼠的神经功能缺陷恢复(P <0.01),减轻脑组织损伤程度。与SAH+vehicle组比较,SAH+LIG20组基底动脉管壁变薄,管径增大(P <0.01),痉挛缓解;基底动脉内皮细胞凋亡减轻,p53、cleaved caspase-3蛋白的表达减少(P <0.05),Bcl-2蛋白的表达增加(P <0.05)。结论:在大鼠SAH模型中,LIG可能通过减少基底动脉内皮细胞的凋亡,缓解脑血管痉挛,从而发挥神经保护作用。Objective: To investigate the effects of Z-Ligustilide(LIG) on delayed cerebral vasospasm(DCVS) after subarachnoid hemorrhage(SAH) in rats.Methods: One hundred SD rats were randomly divided into four groups:Sham group(n = 22),SAH + vehicle group(n = 28),SAH + 5 mg/kg LIG group(SAH + LIG5,n = 25),and SHA +20 mg/kg LIG group(SAH + LIG20,n = 25).The rat SAH model was prepared by modified occipital cisterna secondary blood injection.Thirty minutes after the first blood injection,rats in SAH + LIG5 group and SAH + LIG20 group were injected with 5 mg/kg or 20 mg/kg of LIG in tail vein,while rats in SAH + vehicle group were injected with an equal volume of vehicle(3% Tween 80).The injections were administered once a day for 7 days.The daily neurological function of rats was evaluated according to the Garcia neurobehavioral score.Morphological changes of neurons in rats cortex and hippocampus were detected by Nissl staining.The wall thickness and cross-sectional area of basilar artery(BA) were measured by HE staining to evaluate the condition of vasospasm.Apoptotic endothelial cells of BA were determined by TUNEL staining.The expressions of p53,Bcl-2,Bax,and cleaved caspase-3 in BA were detected by Western Blot and immunohistochemistry.Results: LIG treatment(20 mg/kg) improved neurobehavioral deficits(P <0.01) and alleviated brain damage.Compared with the SAH + vehicle group,the SHA + LIG20 group showed mild vasospasm,which is characterized by reduced wall thickness and increased cross-sectional area of BA(P < 0.01).Moreover,administration of LIG reduced the number of apoptotic endothelial cells of BA,which accompanied downregulation of p53 and cleaved caspase-3 expressions(P < 0.05) and up-regulation of Bcl-2 expression(P < 0.05).Conclusion: LIG may display neuroprotective effect through reducing apoptosis of basal artery endothelial cells,and subsequently relieving cerebral vasospasm in rat SAH model.

关 键 词：蛛网膜下腔出血 迟发性脑血管痉挛 藁本内酯 凋亡 大鼠 

分 类 号：R743.35[医药卫生—神经病学与精神病学]