加味柴胡疏肝汤对急性癫痫小鼠海马miR-187/Wnt/β-catenin信号通路表达的影响  

Effect of Modified Chaihu Shugan Decoction(加味柴胡疏肝汤)on Expression of miR-187/Wnt/β-catenin Signaling Pathway in Hippocampus of Acute Epileptic Mice

作　　者：吕婷婷 李瑞暖 李洁玉 黄其柳 黄小妹 黄柳瑛 周艳英[2] 罗玉珍 梁霜[2] 刁丽梅[2] LYU Tingting;LI Ruinuan;LI Jieyu;HUANG Qiliu;HUANG Xiaomei;HUANG Liuying;ZHOU Yanying;LUO Yuzhen;LIANG Shuang;DIAO Limei(Guangxi University of Chinese Medicine,Nanning 530001,Guangxi,China;First Affiliated Hospital of Guangxi University of Chinese Medicine,Nanning 530023,Guangxi,China)

机构地区：[1]广西中医药大学,广西南宁530001 [2]广西中医药大学第一附属医院,广西南宁530023

出　　处：《中华中医药学刊》2023年第9期89-93,I0027,I0028,共7页Chinese Archives of Traditional Chinese Medicine

基　　金：国家自然科学基金项目(81960858);广西中医脑病临床研究中心项目(桂科AD 20238028);广西研究生教育创新计划项目(YCXJ2021031,YCSW2021225)。

摘　　要：目的探讨加味柴胡疏肝汤对急性癫痫小鼠miR-187/Wnt/β-catenin信号通路表达的影响。方法制备匹罗卡品急性癫痫小鼠模型后,将小鼠随机分为模型组、加味柴胡疏肝汤组、卡马西平组、加味柴胡疏肝汤+miR-187过表达组、加味柴胡疏肝汤+miR-187抑制组,每组10只,除模型组之外,其余组别分别给予相应药物灌胃进行干预,2次/d,连续14 d。脑电图检测小鼠痫性波,苏木素-伊红染色法(hematoxylin-eosin staining,HE)染色法进行正常组与模型组病理组织学观察,实时荧光定量聚合酶链式反应(Real-time PCR,RT-PCR)检测小鼠海马组β-catenin、cyclin D1、c-myc、wnt3 mRNA表达水平,免疫蛋白印迹法(western blot,WB)检测小鼠海马组织β-catenin、cyclin D1、c-myc、wnt3蛋白表达水平,免疫荧光检测小鼠海马组织β-catenin、cyclin D1、c-myc、wnt3的表达水平。结果脑电图结果显示,与正常组相比,模型组小鼠脑电图观察到连续性痫性放电,表现为棘波、尖波、棘-慢综合波;与模型组比较,加味柴胡疏肝汤组与卡马西平组均能减少癫痫小鼠脑电图痫性波次数;与加味柴胡疏肝汤组比较,加味柴胡疏肝汤+miR-187过表达组小鼠脑电图中痫性放电次数明显减少,加味柴胡疏肝汤+miR-187抑制组小鼠脑电图中痫性放电次数明显增加。RT-PCR结果显示,与正常组相比,模型组β-catenin、cyclin D1、c-myc和wnt3基因表达显著升高;与模型组相比,加味柴胡疏肝汤组和加味柴胡疏肝汤+miR-187过表达组小鼠海马组织中β-catenin、cyclin D1、c-myc和wnt3基因表达均明显减低,且较单纯使用加味柴胡疏肝汤组明显减低;加味柴胡疏肝汤+miR-187抑制组cyclin D1较模型组明显降低;卡马西平组β-catenin、cyclin D1、c-myc、wnt3基因表达水平均明显降低。WB结果显示,与正常组相比,模型组β-catenin、cyclin D1、c-myc和wnt3蛋白表达显著升高;与模型组相比,加味柴胡疏肝汤组和卡马西平组Objective To investigate the effect of Modified Chaihu Shugan Decoction(加味柴胡疏肝汤)on the expression of miR-187/Wnt/β-catenin signaling pathway in acute epilepsy mice.Methods After preparing a mouse model of acute epilepsy induced by pilocarpine,the mice were randomly divided into model group,Modified Chaihu Shugan Decoction group,carbamaz-epine group,Modified Chaihu Shugan Decoction+miR-187 overexpression group,Modified Chaihu Shugan decoction+miR-187 inhibition group,with 10 mice in each group.In addition to the model group,the other groups were given the corresponding drugs by gavage twice a day for 14 days.The epileptic wave of mice was detected by electroencephalogram.Hematoxylin-eosin staining(HE)was used to observe the histopathology of normal group and model group.Real-time quantitative polymerase chain reaction(RT-PCR)was used to detect the expression levels ofβ-catenin,cyclin D1,c-myc and wnt3 mRNA in hippocampus of mice.The protein expression levels ofβ-catenin,cyclin D1,c-myc and wnt3 in hippocampus of mice were detected by Western blot(WB).The expression levels ofβ-catenin,cyclin D1,c-myc and wnt3 in hippocampus of mice were detected by immunofluorescence.Results Compared with the normal group,continuous epileptic discharges were observed in the EEG of mice in the model group,which were characterized by spikes,sharp waves and spike slow complex waves.Compared with the model group,the Modified Chaihu Shugan Decoction group and the carbamazepine group could reduce the number of epileptic waves in the EEG of epileptic mice.Compared with the modified Chaihu Shugan Decoction group,the number of epileptic discharges in the EEG of the Modified Chaihu Shugan Decoction+miR-187 overexpression group was significantly reduced,and the number of ep-ileptic discharges in the EEG of the Modified Chaihu Shugan Decoction+miR-187 inhibition group was significantly increased.RT-PCR results showed that compared with the normal group,the expressions ofβ-catenin,cyclin D1,c-myc and wnt3 genes in the model g

关 键 词：加味柴胡疏肝汤 癫痫 海马 miR-187 WNT/Β-CATENIN信号通路 

分 类 号：R285.5[医药卫生—中药学]