海风藤提取物对Aβ_(1-42)寡聚体诱导阿尔茨海默氏病模型鼠认知功能障碍干预的可能机制  

Possible mechanism of intervention of Piper Kadsura Ohwi Extracts on cognitive impairment induced by Aβ_(1-42) oligomer in Alzheimer's disease model rats

作　　者：崔瑶瑶 李雷[2] 隋少梅 王琪 郑梅梅 杜怡峰 贺燕[2] CUI Jao-yao;LI Lei;SUI Shao-mei;WANG Qi;ZHENG Mei-mei;DU Yi-feng;HE Yan(Shandong first Medical University&Shandong Academy of Medical Sciences,Titian,Shandong 271016,China;Department of Neurology,The First Affiliated Hospital of Shandong First Medical University&Shandong Provincial Qianfoshan Hospital,Jinan,Shandong 250014,China;Department of Neurology,Shandong First Medical University Affiliated Provincial Hospital&Shandong Provincial Hospital,Jinan,Shandong 250022,China)

机构地区：[1]山东第一医科大学山东省医学科学院,山东泰安271016 [2]山东第一医科大学第一附属医院山东省千佛山医院神经病学,山东济南250014 [3]山东第一医科大学附属省立医院山东省立医院神经内科,山东济南250022

出　　处：《阿尔茨海默病及相关病杂志》2022年第1期46-52,共7页Chinese Journal of Alzheimer's Disease and Related Disorders

基　　金：国家自然科学基金(30973816);山东省自然科学基金(NO.ZR2015HL120);济南市临床医学科技创新计划(202019158)。

摘　　要：目的:探讨海风藤提取物(kadsura ohwi extract,PKO)对Aβ_(1-42)寡聚体(AβO)诱导阿尔茨海默氏病(Alzheimer disease)模型大鼠认知功能障碍干预的可能机制。方法:健康雄性SD大鼠36只,随机数字表法分为6组:正常对照组(Control组)、假手术组(Sham组)、载体组(高密度脂蛋白和轻乙基哌嗪乙磺酸组)、AβO组、AβO+海风藤组(AβO+PKO组)、AβO+DMSO组。采用微渗泵向侧脑室持续灌注AβO法制作动物模型。应用水迷宫实验观察大鼠的学习记忆功能变化,电镜下观察海马CA1区神经细胞超微结构,免疫印迹法(Western blot)检测NF-κB,TLR4表达,酶标记免疫吸附法(ELISA)检测TNF-α的表达。结果:水迷宫实验结果显示,与Control组、Sham组和载体组比较,AβO组、AβO+DMSO组和AβO+PKO组逃避潜伏期显著延长、穿越平台的次数显著减少、时间比率和路程比率也显著减低(P均<0.05);与AβO组、AβO+DMSO组比较,AβO+PKO组逃避潜伏期显著缩短、穿越平台的次数显著增加、时间比率和路程比率也显著增高(P均<0.05)。电镜下Control组、Sham组、载体组海马CA1区神经细胞超微结构未见明显异常,AβO组和AβO+DMSO组均见到大量凋亡神经细胞,存活的神经细胞线粒体、内质网、终池结构呈现不同程度的损伤。AβO+PKO组可见少量凋亡神经细胞,细胞肿胀轻,细胞器较完整,存活的神经元细胞较多。WB、ELISA结果显示,AβO组、AβO+DMSO组与Control组、Sham组、载体组相比,TLR4、NF-κB、TNF-α表达均显著增多,A0O+PKO组TLR4、NF-κB、TNF-α表达较AβO组、AβO+DMSO组减少(P均<0.05)。偏相关分析显示,TLR4、NF-κB、TNF-α与穿越平台次数均呈负相关(P均=0.00),TLR4、NF-κB、TNF-α之间均为正相关(P均=0.00)。结论:PKO通过降低朋0诱导的TLR4、NF-κB、TNF-α表达,减轻细胞损伤的机制,改善AD模型鼠认知功能。Objective:To explore the possible mechanism of intervention of Kadsura Ohwi extract(PKO)on cognitive impairment in rats with Alzheimer's disease(AD)induced by Aβ_(1-42) oligomer(AβO).Methods:36 healthy male SD rats were randomly divided into 6 groups:normal control group(Control group),sham operation group(Sham group),Carrier group(high density lipoprotein and hydroxyethyl piperazine ethanesulfonic acid group),AβO+carrier group(AβO group),AβO+Kadsura group(AβO+PKO group),AβO+DMSO group.The animal model was made by continuous infiision of AβO into the lateral ventricle by microosmotic pump.The changes of learning and memory function of rats were observed by water maze test,the ultrastructure of neurons in hippocampal CAI region was observed under electron microscope,the expression ofNF-KB and TLR4 was detected by Western blotting(Westemblot),and the expression of TNF-α was detected by enzyme labeled immunosorbent assay(ELISA).Results:The results of water maze test showed that compared with Control group,Sham group and Carrier group,AβO group,AβO+DMSO group and AβO+PKO group significantly prolonged escape latency,significantly decreased the number of times of crossing the platform,and significantly decreased the ratio of time and distance(all P<0.05).Compared with AβO group and Ap+DMSO group,AβO+PKO group significantly shortened the latency of escape,significantly increased the ninnber of times of crossing the platform,and the time ratio and distance ratio were also significantly higher than those ofAβO group and Ap+DMSO group(all P<0.05).under electron microscope,no obvious abnonnality was found in the ultrastructure of hippocampal CAI region in Control group,Sham group and Carrier group.A large number of apoptotic nerve cells were foxind in AβO group and AβO+DMSO group,and the structures of mitochondria,endoplasmic reticulum and terminal cistem of surviving nerve cells were damaged in different degrees.A small number of qx)ptotic nerve cells were found in AβO+PKO group,with mild swelling,intact

关 键 词：阿尔茨海默病 海风藤提取物 Β-淀粉样蛋白 大鼠 认知功能 海马 Toll样受体4 核因子-κB 肿瘤坏死因子-α 

分 类 号：R3[医药卫生—基础医学]