淫黄葛合剂对APP/PS1 HAMP^(-/-)小鼠认知功能和铁死亡氨基酸代谢通路的影响  

Effects of icariin-astragaloside IV-puerarin mixture on cognitive function and ferroptosis amino acid metabolism pathway in APP/PS1 HAMP^(−/−)mice

作　　者：刘珊 贺小平 赵炎 钟健民 张烨华 刘怡铭 李佳璇 董贤慧 LIU Shan;HE Xiaoping;ZHAO Yan;ZHONG Jianmin;ZHANG Yehua;LIU Yiming;LI Jiaxuan;DONG Xianhui(Hebei University of Chinese Medicine,Hebei Key Laboratory of Chinese Medicine Research on Cardio-cerebrovascular Dis eases,Shijiazhuang 050091,China)

机构地区：[1]河北中医药大学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

出　　处：《中国病理生理杂志》2024年第3期502-510,共9页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.81803935);河北省自然科学基金资助项目(No.H2023423009);河北省科技计划项目(No.223777145D);河北省中医药管理局课题(No.2023113);河北省卫健委重点科技研究计划(No.20200129)。

摘　　要：目的:探究淫羊藿苷-黄芪甲苷-葛根素(淫黄葛)合剂(YHG)对铁调素(hepcidin,HAMP)基因敲除的APPswe/PS1dE9(APP/PS1 HAMP^(−/−))小鼠认知功能及铁死亡氨基酸代谢通路的影响。方法:实验分为阴性对照(C57BL/6小鼠)组、APP/PS1组、APP/PS1 HAMP^(−/−)组、APP/PS1+YHG组、APP/PS1 HAMP^(−/−)+YHG组、APP/PS1+地拉罗司(DFX)组和APP/PS1 HAMP^(−/−)+DFX组,每组6只。YHG(淫羊藿苷、黄芪甲苷和葛根素的剂量分别为120、80和80 mg/kg)和DFX(100 mg/kg)灌胃给药,C57组、APP/PS1组和APP/PS1 HAMP^(−/−)组小鼠给予等体积蒸馏水灌胃,每日一次,用药2个月。采用组织铁试剂盒检测小鼠脑组织铁离子含量;透射电镜检测小鼠海马神经元线粒体形态学变化;Morris水迷宫检测小鼠学习记忆能力;免疫荧光法检测各组小鼠脑组织神经元核抗原(NeuN)的含量;生化试剂盒检测小鼠脑组织谷胱甘肽(GSH)表达情况;Western blot检测小鼠脑组织谷氨酸半胱氨酸连接酶催化亚基(GCLC)和谷氨酰胺酶2(GLS2)的表达情况。结果:与C57BL/6小鼠相比,APP/PS1小鼠脑铁含量显著升高(P<0.01),线粒体形态破坏严重,学习记忆能力显著下降(P<0.05),脑组织神经元损伤严重(P<0.01),GSH、GCLC和GLS2表达水平均显著降低(P<0.01);与APP/PS1小鼠相比,APP/PS1 HAMP^(−/−)小鼠脑铁含量显著升高(P<0.01),线粒体形态破坏严重,学习记忆能力显著下降(P<0.05),脑组织神经元损伤严重(P<0.01),GSH、GCLC和GLS2表达水平显著降低(P<0.05);使用YHG和DFX后各组小鼠脑铁含量显著降低(P<0.01),线粒体形态破坏减轻,学习记忆能力显著升高(P<0.05),脑组织神经元损伤减轻(P<0.01),GSH、GCLC和GLS2表达水平显著升高(P<0.05)。结论:YHG可改善APP/PS1 HAMP^(−/−)小鼠学习记忆能力,其机制可能与调节铁死亡氨基酸代谢、增强抗氧化能力有关。AIM:To observe the effect of icariin-astragaloside IV-puerarin mixture(Yin-Huang-Ge mixture,YHG)on cognitive function and ferroptosis amino acid metabolism pathway in hepcidin(HAMP)knockout APPswe/PS1dE9(APP/PS1 HAMP^(−/−))mice.METHODS:The mice were divided into 7 groups:negative control(C57BL/6 mice)group,APP/PS1 group,APP/PS1 HAMP^(−/−)group,APP/PS1+YHG group,APP/PS1 HAMP^(−/−)+YHG group,APP/PS1+de ferasirox(DFX)group,and APP/PS1 HAMP^(−/−)+DFX group,with 6 mice in each group.The YHG and DFX were adminis tered intragastrically,while the mice in C57 group,APP/PS1 group and APP/PS1 HAMP^(−/−)group were given intragastric administration of distilled water,once a day for 2 months.The iron content in mouse brain tissues was detected by tissue iron kit.The morphological changes of the mitochondria in hippocampal neurons were observed by transmission electron microscopy.Morris water maze was used to detect the learning and memory ability of the mice.The content of neuronal nu clear antigen(NeuN)in mouse brain tissues was detected by immunofluorescence staining.The expression of glutathione(GSH)in mouse brain tissues was detected by biochemical kit.The expression levels of glutamate-cysteine ligase catalytic subunit(GCLC)and glutamatase 2(GLS2)in mouse brain tissues were detected by Western blot.RESULTS:Compared with C57BL/6 mice,the brain iron content of APP/PS1 mice was significantly increased(P<0.01),the mitochondria were seriously damaged,the learning and memory ability was significantly decreased(P<0.05),the brain neurons were seri ously damaged(P<0.01),and the expression levels of GSH,GCLC and GLS2 were significantly decreased(P<0.01).Compared with APP/PS1 mice,the brain iron content of APP/PS1 HAMP^(−/−)mice was significantly increased(P<0.01),the mitochondria were seriously damaged,the learning and memory ability was significantly decreased(P<0.05),the brain neurons were seriously damaged(P<0.01),and the expression levels of GSH,GCLC and GLS2 were significantly decreased(P<0.05).After tr

关 键 词：阿尔茨海默病 淫羊藿苷 黄芪甲苷 葛根素 铁死亡 

分 类 号：R742[医药卫生—神经病学与精神病学] R285.5[医药卫生—临床医学] R363.2