地黄饮子通过SIRT2调节突触可塑性改善学习记忆障碍小鼠认知功能的作用  

Dihuang Yinzi Improves Cognitive Function of Mouse Model of Learning and Memory Impairments by Regulating Synaptic Plasticity via SIRT2

作　　者：王文婷 郝阳晶 苏文娜 李钦青 楚世峰 张俊龙 贺文彬 WANG Wenting;HAO Yangjing;SU Wenna;LI Qinqing;CHU Shifeng;ZHANG Junlong;HE Wenbin(Shanxi Key Laboratory of Chinese Medicine Encephalopathy,National International Joint Research Center for Molecular Chinese Medicine,Shanxi University of Chinese Medicine,Jinzhong 030619,China;State Key Laboratory of Bioactive Substances and Functions of Natural Medicines,Institute of Materia Medica&Neuroscience Center,Chinese Academy of Medical Sciences and Peking Union Medical College,Beijing 100050,China)

机构地区：[1]山西中医药大学,分子中医药学国家国际联合研究中心,中医脑病学山西省重点实验室,山西晋中030619 [2]中国医学科学院药物研究所神经科学中心天然药物活性物质与功能国家重点实验室,北京100050

出　　处：《中国实验方剂学杂志》2025年第6期9-17,共9页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(82274388);山西中医药大学研究生创新创业教育项目(X2024CX027)。

摘　　要：目的:研究地黄饮子对东莨菪碱(SCOP)致学习记忆障碍小鼠认知功能的影响并探索其作用机制。方法:采用腹腔注射SCOP诱导学习记忆障碍小鼠模型。将60只雄性C57BL/6J小鼠随机分为正常组(0.9%NaCl,n=10),模型组(SCOP 1 mg·kg^(-1)·d^(-1),n=10),地黄饮子低、中、高剂量组(SCOP 1 mg·kg^(-1)·d^(-1)+地黄饮子5.5、11.0、22.0 g·kg^(-1)·d^(-1),n=10)和多奈哌齐组(SCOP 1 mg·kg^(-1)·d^(-1)+多奈哌齐0.84 mg·kg^(-1)·d^(-1),n=10),对应药物连续干预6周,第4周开始造模,除正常组外,其余各组小鼠于每日灌胃40 min后腹腔注射SCOP造模,第5周开始行为学测试,于每日造模30 min后开始。采用Morris水迷宫、新物体识别实验评估学习记忆障碍小鼠空间学习记忆功能,尼氏染色观察海马CA1区神经元细胞存活数及尼氏小体,蛋白免疫印迹法(Western blot)检测海马沉默信息调节因子2(SIRT2)、α-氨基-3-羟基-5-甲基-4-异恶唑丙酸(AMPA)受体1(GluA1)、蛋白激酶A(PKA)、cAMP反应元件结合蛋白(CREB)、磷酸化(p)-CREB、突触后致密物95(PSD95)、生长相关蛋白-43(GAP-43)、突触素(SYN)的蛋白表达,免疫荧光法检测海马DG区双皮质素(DCX)的蛋白表达情况。结果:与正常组比较,模型组小鼠学习记忆能力显著下降(P<0.01),海马CA1区神经元损伤明显,神经元存活数显著减少(P<0.01),海马DG区DCX蛋白表达显著降低(P<0.01),海马组织GluA1、PKA、p-CREB/CREB、PSD95、SYN、GAP-43蛋白水平明显降低(P<0.05,P<0.01)、SIRT2蛋白水平显著增加(P<0.01);与模型组比较,地黄饮子中、高剂量组和多奈哌齐组学习记忆能力明显改善(P<0.05,P<0.01),地黄饮子低、中、高剂量组和多奈哌齐组神经元存活数显著增加(P<0.01),地黄饮子中剂量组和多奈哌齐组DCX蛋白表达明显增加(P<0.05,P<0.01),地黄饮子中、高剂量组和多奈哌齐组GluA1、PKA、p-CREB/CREB、PSD95、SYN、GAP-43蛋白水平明显上调(P<0.05,P<0.01),SIRT2蛋白水平显�Objective:To investigate the effects of Dihuang Yinzi on the cognitive function in the mouse model of learning and memory impairments induced by scopolamine(SCOP)and explore the treatment mechanism.Methods:A mouse model of learning and memory impairment was induced by intraperitoneal injection of SCOP.Sixty male C57BL/6J mice were randomized into six groups:control(0.9%NaCl,n=10),model(SCOP 1 mg·kg^(-1)·d^(-1),n=10),low-,medium-,and high-dose Dihuang Yinzi(SCOP 1 mg·kg^(-1)·d^(-1)+Dihuang Yinzi 5.5,11.0,and 22.0 g·kg^(-1)·d^(-1),n=10),and donepezil(SCOP 1 mg·kg^(-1)·d^(-1)+donepezil 0.84 mg·kg^(-1)·d^(-1),n=10).Mice were administrated with corresponding drugs for 6 weeks.Modeling started in the 4th week,and mice in other groups except the control group were injected with SCOP intraperitoneally 40 min after daily gavage.Behavioral testing began in the 5th week,30 min after modeling each day.The Morris water maze and novel object recognition tests were carried out to evaluate the spatial learning and memory function of mice.Nissl staining was employed to observe the survival of neurons and Nissl bodies in the hippocampal CA1 region.Western blot was employed to determine the protein levels of silent information regulator 2(SIRT2),α-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid(AMPA)receptor 1(GluA1),protein kinase A(PKA),cAMP response element-binding protein(CREB),phosphorylated-CREB(p-CREB),postsynaptic density protein 95(PSD95),growth-associated protein-43(GAP-43),and synaptophysin(SYN)in the hippocampus.Immunofluorescence was used to detect the expression of doublecortin(DCX)in the hippocampal dentate gyrus(DG)region.Results:Compared with the control group,the model group showed impaired learning and memory(P<0.01),obvious neuronal damage in the hippocampal CA1 region,a reduction in neuron survival(P<0.01),a decrease in DCX expression in the hippocampal DG region(P<0.01),down-regulated proteins levels of GluA1,PKA,p-CREB/CREB,PSD95,SYN,and GAP-43 in the hippocampal tissue(P<0.05,P<0.01),and an up

关 键 词：地黄饮子 学习记忆障碍 蛋白激酶A(PKA)/cAMP反应元件结合蛋白(CREB)信号通路 沉默信息调节因子2 突触可塑性 

分 类 号：R285[医药卫生—中药学] R289[医药卫生—中医学] R259