益智清心方调控线粒体分裂融合改善阿尔茨海默病小鼠认知功能的机制研究  

Study on Mechanism of Yizhi Qingxin Formula Regulating Mitochondrial FissionFusion on Improving Cognitive Function of Alzheimer's Disease Mice

作　　者：李泽惠 张业昊 曹宇[1] 裴卉[1] 马丽娜[1] 杨洋[1] 李浩[1,2] LI Zehui;ZHANG Yehao;CAO Yu;PEI Hui;MA Lina;YANG Yang;LI Hao(Xiyuan Hospital,China Academy of Chinese Medical Sciences,Beijing 100091,China;Wangjing Hospital,China Academy of Chinese Medical Sciences,Beijing 100102,China)

机构地区：[1]中国中医科学院西苑医院,北京100091 [2]中国中医科学院望京医院,北京100102

出　　处：《中国中医基础医学杂志》2023年第7期1096-1102,共7页JOURNAL OF BASIC CHINESE MEDICINE

基　　金：国家自然科学基金项目(82205103,82205233);中国中医科学院基本科研业务费优秀青年科技人才培养专项(ZZ15-YQ-018);国家重点研发计划“中医药现代化”重点专项(2022YFC3501400)。

摘　　要：目的 研究益智清心方(Yizhi Qingxin formula,YQF)改善认知功能和调控线粒体分裂/融合的机制。方法 采用APPswe/PS1De9(APP/PS1)双转基因小鼠为阿尔茨海默病(Alzheimer disease,AD)模型,随机分为模型组、YQF 2.6 g/kg组、YQF 5.2 g/kg组、多奈哌齐组、PRE-084组和YQF 5.2 g/kg+BD1047组,以C57BL/6J小鼠为对照组。连续干预8周后,水迷宫实验检测小鼠的认知功能,免疫组化法检测小鼠海马区的β-淀粉样蛋白(β-amyloid,Aβ)沉积,透射电镜观察和定量分析小鼠海马神经元内线粒体的数量和长轴长度,免疫荧光检测sigma-1受体(sigma-1 receptor,Sig-1R)与神经元核蛋白(neuronal nuclear antigen,Neu N)的共定位情况,Western blot法检测海马组织动力相关蛋白1(dynamin-related protein 1,DRP1)、磷酸化DRP1(phospho-DRP1,p-DRP1)(Ser616)、分裂蛋白1(fission 1,Fis1)、线粒体融合蛋白2(mitofusin 2,MFN2)、视神经萎缩蛋白1(optic atrophy 1,OPA1)以及Sig-1R的表达。结果与模型组比较,YQF 5.2 g/kg组小鼠的水迷宫逃避潜伏期缩短(P<0.05),穿台次数增多(P<0.05);Aβ沉积减少(P<0.01);海马神经元线粒体数量减少(P<0.05),线粒体长轴长度延长(P<0.01);Fis1表达下降(P<0.05),OPA1和MFN2的表达升高(P<0.01);Sig-1R表达升高(P<0.01),Sig-1R与Neu N的共定位增多(P<0.01)。且以上作用可以被Sig-1R抑制剂BD1047抑制。结论 YQF能够恢复海马神经元线粒体分裂和融合的平衡,从而改善APP/PS1小鼠的认知功能,其机制与Sig-1R激活有关。Objective To investigate the mechanism of Yizhi Qingxin formula(YQF)in improving cognitive function and regulating mitochondrial fission/fusion.Methods APP/PS1 double transgenic mice were used as Alzheimer disease,(AD)models and randomly divided into model group,YQF 2.6 g/kg group,YQF 5.2 g/kg group,donepezil group,PRE-084 group,and YQF 5.2 g/kg+BD1047 group.C57BL/6J mice were used as a control group.After 8 weeks of continuous intervention,the cognitive function of mice was tested by water maze test.β-amyloid(Aβ)deposition in the hippocampus of mice was detected by immunohistochemistry.The number and length of mitochondria in the hippocampal neurons of mice were observed by transmission electron microscopy.The co-localization of Sigma-1 receptor(Sig-1R)and neuronal nuclear antigen(NeuN)was detected by immunofluorescence.The expressions of dynamin-related protein 1(DRP1),phospho-DRP1(p-DRP1)at Ser616,fission 1(Fis1),mitofusin 2(MFN2),optic atrophy 1(OPA1),and Sig-1R in the hippocampus were detected by Western blot.Results Compared with the model group,the water maze escape latency of the YQF 5.2 g/kg group was shortened(P<0.05),the number of times of crossing the platform was increased(P<0.05),the Aβdeposition decreased(P<0.01).The number of mitochondria in neurons was decreased(P<0.05),and the length of of mitochondria was prolonged(P<0.01).The expression of Fis1 was decreased(P<0.05),and the expression of OPA1 and MFN2 was increased(P<0.01).The expression of Sig-1R protein was increased(P<0.01),and the colocalization of Sig-1R and NeuN was increased(P<0.01).And the above effects can be inhibited by the Sig-1R inhibitor BD1047.Conclusion YQF can restore the balance of mitochondrial fission and fusion in hippocampal neurons,thereby improving the cognitive function of APP/PS1 mice,and the mechanism is related to the activation of Sig-1R.

关 键 词：阿尔茨海默病 益智清心方 sigma-1受体 线粒体分裂和融合 

分 类 号：R285[医药卫生—中药学]