益气活血方对脑缺血再灌注大鼠学习记忆能力及海马突触可塑性的影响  

Effects of Yiqi-Huoxue decoction on learning,memory and hippocampal synaptic plasticity in cerebral ischemia/reperfusion rats

作　　者：李佩佩[1,2] 时潇[1,2] 王瑞 王又闻[3] 王丽娜[1] 胡建鹏[1,2] LI Pei-pei;SHI Xiao;WANG Rui;WANG You-wen;WANG Li-na;HU Jianpeng(Key Laboratory of Xin'an Medical Ministry of Education,Anhui University of Traditional Chinese Medicine,Hefei 230012,China;Graduate School of Anhui University of Traditional Chinese Medicine,Hefei 230012,China;School of Pharmacy,Shanghai University of Chinese Medicine,Shanghai 201203,China)

机构地区：[1]安徽中医药大学新安医学教育部重点实验室,安徽合肥230012 [2]安徽中医药大学中医学院,安徽合肥230012 [3]上海中医药大学药学院,上海201203

出　　处：《中国病理生理杂志》2022年第12期2249-2257,共9页Chinese Journal of Pathophysiology

基　　金：安徽省第十三批115产业创新团队资助项目(皖人才办【2020】4号);安徽省自然科学基金面上项目(No.2208085MH265);国家中医药管理局学术流派传承工作室第二轮建设项目(国中医药办人教函[2019]62号);安徽中医药大学2021年度科技创新基金重点项目(No.2021ZD02)。

摘　　要：目的:探讨益气活血方(YQHXD)对脑缺血再灌注(I/R)大鼠学习记忆功能,海马区突触超微结构及脑源性神经营养因子(BDNF)、突触小泡蛋白(SYN)和突触后致密蛋白95(PSD95)蛋白的影响。方法:采用改良线栓法制备大脑中动脉阻塞再灌注(MCAO/R)大鼠模型,将造模成功大鼠随机分为模型组(I/R组)及YQHXD低、中、高剂量组,并设置假手术(sham)组作为对照,每组12只。YQHXD低、中、高剂量组分别按照4.2、8.4和16.8 g·kg^(-1)·d^(-1)的剂量灌胃,sham组和I/R组给予等体积生理盐水,于用药14 d后采用Longa评分法观察各组大鼠神经功能缺损情况,Morris水迷宫实验评估各组大鼠学习与记忆功能,苏木素-伊红(HE)染色观察缺血侧海马CA1区及齿状回(DG)病理变化,透射电镜观察海马区突触数量、突触后致密物(PSD)厚度和结构变化,Western blot检测海马组织BDNF、SYN和PSD95蛋白表达水平。结果:与sham组比较,I/R组大鼠神经功能缺损评分显著增高(P<0.01),水迷宫逃避潜伏期显著延长(P<0.01),平台穿越次数显著减少(P<0.05),海马CA1区和DG区病理损伤严重,海马区突触超微结构散乱,突触数密度显著减少(P<0.01),PSD厚度显著变薄(P<0.01),BDNF、SYN和PSD95蛋白表达显著下降(P<0.01);与I/R组相比,YQHXD各组均能改善模型大鼠神经功能缺损情况,修复海马区组织病理损伤和突触超微结构;其中YQHXD中、高剂量组能显著减少水迷宫逃避潜伏期(P<0.01),显著提高平台穿越次数(P<0.05),显著增加突触数密度和PSD厚度(P<0.05),同时提高BDNF、SYN和PSD95蛋白表达水平。结论:YQHXD可改善MCAO/R大鼠神经功能,提高学习记忆能力,减轻海马神经元损伤,修复突触结构,提高突触数密度和PSD厚度,其作用机制可能与促进缺血侧海马组织BDNF、SYN和PSD95突触相关蛋白表达,提高突触可塑性有关。AIM:To investigate the effect of Yiqi-Huoxue decoction(YQHXD)on learning and memory ability of rats with cerebral ischemia/reperfusion(I/R).METHODS:Middle cerebral artery occlusion/reperfusion(MCAO/R)model in rats was prepared by the modified suture method,and the successful model rats were randomly divided into model group(I/R group),and low-,medium-and high-dose YQHXD(YQHXD-low,-medium and-high)groups.Sham operation(sham)group served as control.There were 12 rats in each group.The rats in YQHXD-low,-medium and-high groups were given 4.2,8.4 and 16.8 g·kg^(-1)·d^(-1)YQHXD by intragastric administration,respectively.The rats in sham group and I/R group were given an equal volume of normal saline.After 14 d of administration,the Longa score was used to observe the neurological deficits of the rats in each group.Morris water maze test was used to evaluate the learning and memory function of the rats in each group.Hematoxylin-eosin(HE)staining was used to observe the pathological changes in CA1 area and dentate gyrus(DG)of the ischemic hippocampus.The number of synapses,postsynaptic density(PSD)thickness,and structural changes in the hippocampus were observed by transmission electron microscopy.The protein expression levels of brain-derived neurotrophic factor(BDNF),synaptophysin(SYN)and PSD95 in the hippocampus were detected by Western blot.RESULTS:Compared with sham group,the neurological deficit score of the rats in I/R group was significantly increased(P<0.01),the water maze escape latency was significantly prolonged(P<0.01),and the number of platform crossings was significantly decreased(P<0.05).The pathological damage in the CA1 and DG area was severe,the synaptic ultrastructure in the hippocampus was scattered,the synaptic numerical density was significantly reduced(P<0.01),the PSD thickness was significantly thinner(P<0.01),and the protein expression levels of BDNF,SYN and PSD95 were significantly decreased(P<0.01).Compared with I/R group,each dose of YQHXD attenuated the neurological deficits and the hist

关 键 词：益气活血方 脑缺血再灌注损伤 突触可塑性 学习记忆能力 脑源性神经营养因子 

分 类 号：R743.3[医药卫生—神经病学与精神病学] R965[医药卫生—临床医学]