淫羊藿、黄芪、葛根有效组分复方介导铁调素干预AD细胞模型ADAM10表达的研究  

Effect of compound of Epimedium,Astragalus and Radix Puerariae on expression of ADAM10 in Aβ-induced hippocampal neuron HT22 cells with or without hepcidin expression knock-down

作　　者：马冬雪 高维娟[1] 刘真一[1] 张红军 王嘉颖 董贤慧 MA Dong-xue;GAO Wei-juan;LIU Zhen-yi;ZHANG Hong-jun;WANG Jia-ying;DONG Xian-hui(Hebei Key Laboratory of Chinese Medicine Research on Cardio-cerebrovascular Disease,Hebei University of Chinese Medicine,Shjiazhuang 050091,China;Department of Pathophysiology,Chengde Medical College,Chengde 067000,China)

机构地区：[1]河北中医学院,河北省心脑血管病中医药防治研究重点实验室,河北石家庄050091 [2]承德医学院病理生理学教研室,河北承德067000

出　　处：《中国病理生理杂志》2020年第12期2198-2204,共7页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金青年项目(No.81803935);河北省自然基金面上项目(No.H2019423095);河北省教育厅青年拔尖人才项目(No.BJ2018059);河北省卫健委重点科技研究计划(No.20200129);河北省中医药管理局课题(No.2017081);承德市科技计划项目(No.201601A022)。

摘　　要：目的:探讨淫羊藿、黄芪、葛根有效组分复方介导铁调素(hepcidin,HAMP)对阿尔茨海默病(Alzheimer disease,AD)细胞模型淀粉样前体蛋白水解中关键酶去整合素金属蛋白酶10(a disintegrin and metalloproteinase 10,ADAM10)表达的影响。方法:体外培养小鼠海马神经元细胞系HT22,随机分为7组:正常对照组、Aβ组(Aβ25-35诱导HT22细胞建立AD细胞模型)、RNAi组(沉默HAMP基因)、Aβ+RNAi组(建立AD细胞模型并沉默HAMP基因)和Aβ+TCM组(建立AD细胞模型并应用淫羊藿、黄芪、葛根有效组分复方处理)、RNAi+TCM组(沉默HAMP基因并应用淫羊藿、黄芪、葛根有效组分复方处理)、Aβ+RNAi+TCM组(建立AD细胞模型沉默HAMP基因并应用淫羊藿、黄芪、葛根有效组分复方处理)。应用qPCR和Western blot检测HAMP的沉默效率应用免疫荧光、qPCR和Western blot检测各组ADAM10的表达情况。结果:与正常对照组比较,Aβ组、RNAi组和Aβ+RNAi组中的ADAM10表达显著降低(P<0.05);与Aβ组比较,Aβ+RNAi组中ADAM10表达显著降低(P<0.05),Aβ+TCM组中ADAM10表达显著增高(P<0.05);与RNAi组比较,Aβ+RNAi组中的ADAM10表达显著降低(P<0.05),RNAi+TCM组中的ADAM10表达显著增高(P<0.05);与Aβ+RNAi组比较,Aβ+RNAi+TCM组中的ADAM10表达显著增高(P<0.05)。结论:淫羊藿、黄芪、葛根有效组分复方可上调AD细胞模型ADAM10的表达,其机制可能与HAMP表达有关。AIM:To explore the effect of compound of Epimedium,Astragalus and Radix Puerariae on the expression of a disintegrin and metalloproteinase 10(ADAM10)in Aβ-induced hippocampal neuron HT22 cells with or without hepcidin(HAMP)expression knock-down for analyzing the pathogenesis of Alzheimer disease(AD)at cell level.METHODS:Hippocampal neuron HT22 cells were cultured in vitro and randomly divided into 7 groups:control group,Aβgroup(Aβ25-35-induced HT22 cells),RNAi group(HAMP gene was silenced in HT22 cells),Aβ+RNAi group(HAMP gene expression in Aβ25-35-induced HT22 cells was silenced),Aβ+TCM group(Aβ25-35-induced HT22 cells were treated with Epimedium,Astragalus root and Radix Puerariae effective components),RNAi+TCM group(HT22 cells with HAMP gene silence were treated with Epimedium,Astragalus root and Radix Puerariae effective components)and Aβ+RNAi+TCM group(Aβ25-35-induced HT22 cells with HAMP gene silence were treated with Epimedium,Astragalus root and Radix Puerariae effective components).The silence efficiency of HAMP siRNA was detected by qPCR and Western blot.The ADAM10 expression in each group was determined by immunofluorescence,qPCR and Western blot.RESULTS:The HAMP siRNA-3 sequence had the highest interference efficiency.Compared with control group,the expression levels of ADAM10 in Aβgroup,RNAi group and Aβ+RNAi group were decreased(P<0.05).Compared with Aβgroup,the expression levels of ADAM10 in Aβ+RNAi group was also decreased(P<0.05),and the expression levels of ADAM10 in Aβ+TCM group was increased(P<0.05).Compared with RNAi group,the expression levels of ADAM10 in Aβ+RNAi group was decreased(P<0.05),while the expression levels of ADAM10 in RNAi+TCM group was increased(P<0.05).Compared with Aβ+RNAi group,the expression levels of ADAM10 in Aβ+RNAi+TCM group was increased(P<0.05).CONCLUSION:The effective components of Epimedium,Astragalus and Radix Puerariae compound promotes the expression of ADAM10 in Aβ25-35-induced HT22 cells,which mechanism may be related to the expression of HAM

关 键 词：阿尔茨海默病 淫羊藿 黄芪 葛根 铁调素 HT22细胞 

分 类 号：R749.16[医药卫生—神经病学与精神病学] R363.2[医药卫生—临床医学]