柴胡疏肝汤对颞叶癫痫小鼠的脑保护作用  

Neuroprotective effects of Chaihu Shugan Decoction on mice with temporal lobe epilepsy

作　　者：朱晓煜 聂玲辉 丁月文 梁小珊[1] 莫孙炼[1] 伍志勇[1] 谢炜[1] ZHU Xiao-yu;NIE Ling-hui;DING Yue-wen;LIANG Xiao-shan;MO Sun-lian;WU Zhi-yong;XIE Wei(College of Traditional Chinese Medicine,Southern Medical University,Guangzhou 510515,Guangdong Province,China;Guangdong Traditional Medical and Sports Injury Rehabilitation Research Institute,Guangzhou 510317,Guangdong Province,China)

机构地区：[1]南方医科大学中医药学院,广州510515 [2]广东省第二人民医院传统医学与运动伤害康复研究所,广东广州510317

出　　处：《中国临床药理学杂志》2021年第10期1158-1162,共5页The Chinese Journal of Clinical Pharmacology

基　　金：国家自然科学基金资助项目(81703926,81873158);广东省自然科学基金资助项目(2017A030310289);广州市科技计划基金资助项目(2018-1002-SF-0635)。

摘　　要：目的研究柴胡疏肝汤对颞叶癫痫小鼠海马区星形胶质细胞活化、星形细胞上调基因-1 (AEG-1) mRNA表达、兴奋性氨基酸转运蛋白2 (EAAT2) mRNA表达的影响,探究柴胡疏肝汤抗癫痫的可能机制。方法将造模成功的慢性颞叶癫痫小鼠随机分为模型组(n=10),丙戊酸钠组(n=10),柴胡疏肝汤高剂量组(n=10),柴胡疏肝汤中剂量组(n=10),柴胡疏肝汤低剂量组(n=10),另取10只小鼠为对照组。丙戊酸钠组小鼠给予丙戊酸钠0.2 g·kg^(-1)灌胃,柴胡疏肝汤高、中、低剂量组分别予柴胡疏肝汤60,30,15 g·kg^(-1)灌胃,对照组与模型组小鼠给予等容积生理盐水灌胃,每天2次,连续干预28 d。同时进行行为学视频监测。用苏木素-伊红(HE)染色、尼氏染色观察小鼠脑组织海马区病变;以免疫荧光法检测小鼠脑组织星形胶质细胞活化情况;以酶联免疫吸附法检测小鼠血清白细胞介素-1β(IL-1β)表达量的变化;以实时荧光定量聚合酶链反应(Real-time PCR)检测海马组织AEG-1、EAAT2基因相对表达水平。结果与对照组比较,模型组小鼠海马CA1、CA3区神经元细胞数量减少,排列紊乱,细胞肿胀破裂,胞浆内尼氏体数量减少,胞核边聚、固缩、核仁消失;海马区GFAP阳性细胞明显增多。对照组、模型组、丙戊酸钠组和柴胡疏肝汤高、中、低剂量组小鼠血清IL-1β浓度分别为(34.00±10.36),(62.18±9.55),(34.46±11.75),(24.37±3.65),(30.82±5.65),(29.09±3.47)pg·mL^(-1),与对照组比较,模型组血清IL-1β表达量和海马组织AEG-1 mRNA表达升高(P<0.05);而海马组织EAAT2 mRNA表达降低(P<0.05)。与模型组比较,丙戊酸钠和柴胡疏肝汤高、中剂量组小鼠海马组织CA1和CA3区的病理改变明显改善,海马区GFAP阳性细胞明显减少,血清IL-1β表达量及海马组织AEG-1 mRNA表达降低(均P<0.05),而海马组织EAAT2 mRNA表达升高(P<0.05)。结论柴胡疏肝汤可以抑制星形胶质细胞活化,下调AEG-1 mRNA的表达,上�Objective To study the effects of Chaihu Shugan Decoction(CHSGD) on both the activation of astrocytes and the expression of astrocyte elevated gene-1(AEG-1) and excitatory acid transporter 2(EAAT2) in hippocampus region of mice with temporal lobe epilepsy, in order to explore the possible antiepileptic mechanism of Chaihu Shugan Decoction. Methods Mouse models with chronic temporal lobe epilepsy were randomly divided into model group(n=10), sodium valproate group(n=10) and CHSGD high(n=10), medium(n=10), low dose ( n = 10) groups,another 10 normal mice were taken as control group. Mice in sodium valproate group were administered intragastrically with 0. 2 g·kg^(-1) sodium valproate;mice in CHSGD high,medium and low dose groups were administered intragastrically with 60,30 and 15 g·kg-1 CHSGD,mice in normal control group and model group were administered intragastrically with equal volume of normal saline,twice a day,for continuous intervention for 28 d.Behavioral video monitoring was conducted simultaneously. Hematoxylin and eosin( HE) stain and Nissl stain were used to observe pathological changes in hippocampus region of mice. Immunofluorescence was used to detect the activation of astrocytes. Enzyme-linked immunosorbent assay( ELISA) was used to measure the expression of interleukin-1β( IL-1β) in serum. Real-time quantitative fluorescence PCR was used to detect the relative expression levels of AEG-1 mR NA and EAAT2 mR NA in hippocampus region. Results Compared to control group,in the hippocampal CA1 and CA3 regions of model group,the number of neurons was decreased,the cell arrangement was disordered and the morphology was swollen,the number of Nissl bodies in cytosol was decreased,chromatin agglutination and pyknosis and nucleolus disappeared. GFAP positive cells increased significantly in hippocampus region. Serum IL-1β expression level in control group,model group,sodium valproate group and CHSGD high,medium and low dose groups were( 34. 00 ± 10. 36),( 62. 18 ± 9. 55),( 34. 46 ± 11. 75),( 24. 37 ±

关 键 词：颞叶癫痫 柴胡疏肝汤 白细胞介素-1Β 星形胶质细胞 星形细胞上调基因-1 兴奋性氨基酸转运蛋白2 

分 类 号：R28[医药卫生—中药学]