芪仙通络方对脑梗死大鼠突触可塑性与功能恢复的影响及机制研究  

Effect and Mechanism of Qixian Tongluo Formula on Synaptic Plasticity and Functional Recovery in Rats with Cerebral Infarction

作　　者：周胜强 寿雅琨[2] 王琦 李博 刘芳 邓奕辉[2] ZHOU Shengqiang;SHOU Yakun;WANG Qi;LI Bo;LIU Fang;DENG Yihui(Hunan Academy of Chinese Medicine,Changsha 410006 Hunan,China;Hunan University of Chinese Medicine,Changsha 410208 Hunan,China;The First Affillated Hospital of Hunan University of Chinese Medicine,Changsha 410007 Hunan,China)

机构地区：[1]湖南省中医药研究院,湖南长沙410006 [2]湖南中医药大学,湖南长沙410208 [3]湖南中医药大学第一附属医院,湖南长沙410007

出　　处：《中药新药与临床药理》2021年第7期912-919,共8页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：国家中医药管理局“刘祖贻国医大师传承工作室建设项目”(2015年);湖南省教育厅重点项目(18A225);湖南省教育厅优秀青年项目(18B253);湖南省中医药管理局一般课题(2021083);湖南省中医药研究院院级重点课题(201902);湖南省“国内一流培育学科”中西医结合开放基金项目(2019ZXYJH01)。

摘　　要：目的观察芪仙通络方对脑梗死大鼠突触可塑性与功能恢复的影响,从脑源性神经营养因子(BDNF)/酪氨酸激酶受体B(TrkB)/环磷酸腺苷应答元件结合蛋白(CREB)信号通路角度探讨其作用机制。方法线栓法复制大脑中动脉阻塞(MCAO)大鼠模型。将36只模型复制成功的大鼠随机分为模型组、芪仙通络方组(15.66 g·kg^(-1),灌胃给药)、K252a组(50μg·kg^(-1),腹腔注射)、芪仙通络方(15.66 g·kg^(-1),灌胃给药)+K252a(50μg·kg^(-1),腹腔注射)组、胞磷胆碱组(0.054 g·kg^(-1),灌胃给药)以及溶剂对照组,另设假手术组6只。术后第3天开始干预给药,每日1次,连续干预12 d。于术后第3、7、14天采用Longa评分法评定大鼠神经功能;术后第14天采用镀银染色和透射电镜分别观察缺血周边组织神经元突触一般结构与超微结构变化,免疫组化法检测缺血周边组织中突触可塑性标志蛋白突触素(SYN)、突触后致密物-95(PSD-95)表达,Western Blot法检测缺血周边组织中BDNF、TrkB、CREB以及磷酸化环磷酸腺苷应答元件结合蛋白(p-CREB)表达。结果与假手术组比较,模型组大鼠神经功能缺损症状明显,各时间点神经功能评分明显升高(P<0.01);缺血周边组织神经元树突分支数量、树突棘与突触数量明显减少,突触结构模糊,部分突触囊泡出现膨大破损,突触后致密物质减少;突触可塑性标志蛋白SYN与PSD-95表达均明显减少(P<0.01);BDNF与p-CREB蛋白表达均增加(P<0.05),而TrkB与CREB蛋白表达相当(P>0.05)。与模型组比较,芪仙通络方组第14天神经功能评分明显降低(P<0.01);缺血周边组织神经元树突分支数量、树突棘与突触数量明显增加,突触结构较为清晰,突触后致密物质与突触囊泡数目增多;SYN、PSD-95、BDNF与p-CREB蛋白表达均明显增加(P<0.05,P<0.01),但TrkB与CREB蛋白表达相当(P>0.05)。与芪仙通络方组比较,芪仙通络方+K252a组第14天神经功能评分有所升高(P<0.01);Objective To observe the effect of Qixian Tongluo formula on synaptic plasticity and functional recovery in rats with cerebral infarction and explore the mechanism of action from the perspective of brain derived neurotrophic factor(BDNF)/tyrosine kinase receptor B(TrkB)/cAMP response element binding protein(CREB)signaling pathway.Methods The middle cerebral artery occlusion(MCAO)rat model was established by suture method.Thirty-six model rats were randomly divided into model group,Qixian Tongluo formula group(15.66 g·kg^(-1),intragastric administration),K252a group(50μg·kg^(-1),intraperitoneal injection),Qixian Tongluo formula(15.66 g·kg^(-1),intragastric administration)+K252a(50μg·kg^(-1),intraperitoneal injection)group,Citicoline group(0.054 g·kg^(-1),intragastric administration)and solvent control group.In addition,six rats underwent sham operation.The intervention administration was started on the 3rd day after operation once a day for 12 consecutive days.On the 3rd,7th,and 14th day after operation,neurological function was evaluated by Longa score.On the 14th day after operation,the general structure and ultrastructural changes of synapse in the peri-ischemia tissue were observed by silver staining and transmission electron microscope,respectively.The expression of synaptic plasticity marker protein[synaptophysin(SYN)and postsynaptic density-95(PSD-95)]in the peri-ischemia tissue were detected by immunohistochemistry,and the expression of BDNF,TrkB,CREB and phosphorylated cAMP response element binding protein(p-CREB)in the peri-ischemia tissue were detected by Western Blot.Results Compared with the sham operation group,model group had obvious symptoms of neurological deficits,the neurological function scores were significantly increased at each time point(P<0.01),the number of neuronal dendritic branches,dendritic spines and synapses in the peri-ischemia tissue were significantly reduced,the synapse structure was blurred,some synaptic vesicles were enlarged and damaged,the post-synaptic dense material

关 键 词：芪仙通络方 脑梗死 突触可塑性 BDNF/TrkB/CREB信号通路 大鼠 

分 类 号：R285.5[医药卫生—中药学]