异功散通过调控脑水液代谢改善APP/PS1转基因小鼠的学习记忆能力  

Yigong San improves learning and memory functions of APP/PS1 transgenic mice by regulating brain fluid metabolism

作　　者：曾静 花雷 阳勇[1,2] 张小梅 魏江平[1,2] 李利生 ZENG Jing;HUA Lei;YANG Yong;ZHANG Xiaomei;WEI Jiangping;LI Lisheng(Sichuan-Chongqing Joint Key Laboratory of Innovation of New Drugs of Traditional Chinese Medicine;Third Level Laboratory of Traditional Chinese Medicine Chemistry of the National Administration of Traditional Chinese Medicine,Chongqing Academy of Chinese Materia Medica,Chongqing 400065,China;Key Laboratory of Basic Pharmacology of Ministry of Education&Ministry of Education International Cooperation Joint Laboratory of Characteristic Ethnic Medicine,Zunyi Medical University,Zunyi 563000,China;Department of Pharmacology&Guizhou Provincial Key Laboratory of Basic Pharmacology,Zunyi Medical University,Zunyi 563000,China)

机构地区：[1]中药新药创制川渝共建重点实验室 [2]国家中医药管理局中药化学三级实验室,重庆市中药研究院,重庆400065 [3]遵义医科大学基础药理教育部重点实验室暨特色民族药教育部国际合作联合实验室,贵州遵义563000 [4]遵义医科大学贵州省基础药理重点实验室//药学院药理学教研室,贵州遵义563000

出　　处：《南方医科大学学报》2024年第10期2015-2023,共9页Journal of Southern Medical University

基　　金：重庆市技术创新与应用发展专项重点项目(cstc2021jscxdxwtBX0012);重庆市自然科学基金项目博士后项目(cstc2021jscxbshX0054);重庆市中药研究院基本科研业务费项目(jbky20210003)。

摘　　要：目的观察异功散对APP/PS1小鼠学习记忆能力的影响,并从调控脑水液代谢系统探讨异功散抗痴呆的作用机制。方法将3月龄雄性APP/PS1转基因小鼠及同龄同背景野生型C57BL/6小鼠随机分为4组(8只/组):对照组、模型组、多奈哌齐(1.67 mg/kg)组、异功散(7.5 g/kg)组。灌胃给予各组对应药物1次/d,1月后采用Morris水迷宫实验考察小鼠的学习记忆能力;HE染色观察海马皮层组织病理形态学变化;免疫组化染色、硫磺素S染色观察脑内淀粉样蛋白斑块数量变化;ELISA法检测脑组织和血清Aβ_(1-40)和Aβ_(1-42)含量;伊文思蓝法检测血脑屏障(BBB)通透性变化;免疫荧光共定位考察AQP4在星型胶质细胞上极化情况;Western blotting观察血管内皮钙黏蛋白(VE-cadherin)、闭锁连接蛋白1(ZO-1)、闭合蛋白(Occludin)、β-淀粉样前体蛋白(APP)、β-分泌酶(BACE1)、胰岛素降解酶(IDE)、低密度脂蛋白受体相关蛋白1(LRP1)、晚期糖基化终末产物受体(RAGE)、水通道蛋白4(AQP4)蛋白表达情况。结果与对照组比较,APP/PS1小鼠第5天的逃避潜伏期显著延长(P<0.001),平台象限停留时间降低(P<0.05);海马和皮层神经细胞变性或坏死细胞数量增加且病理评分升高(P<0.001);Aβ阳性斑块明显升高以及硫磺素S荧光强度明显增强(P<0.05);脑组织和血清Aβ_(1-40)、Aβ_(1-42)含量升高(P<0.05);BBB通透性增加(P<0.01),RAGE蛋白表达上调(P<0.01)而VE-cadherin、LRP1、ZO-1、Occludin、AQP4蛋白表达下调(P<0.05),且AQP4在GFAP阳性细胞上表达的数量减少(P<0.05)。与模型组比较,异功散组小鼠第5天的逃避潜伏期缩短(P<0.001),平台象限停留时间增加(P<0.05)且平均游泳速度加快(P>0.05);海马和皮层神经细胞变性或坏死细胞数量减少且病理评分降低(P<0.01);Aβ阳性斑块减少以及硫磺素S荧光强度减弱(P<0.05);脑组织和血清Aβ_(1-40)、Aβ_(1-42)含量降低(P<0.05);维持了BBB通透性(P<0.01),RAGE蛋白表达下调(PObjective To explore the mechanism by which Yigong San(YGS)improves learning and memory abilities of APP/PS1 transgenic mice in light of cerebral fluid metabolism regulation.Methods Three-month-old male APP/PS1 transgenic mice and wild-type C57BL/6 mice were both randomized into control group,model group,donepezil(1.67 mg/kg)group,and YGS(7.5 g/kg)group and received the corresponding treatments via gavage once daily for one month.After the treatments,the mice were assessed for learning and memory functions using Morris water maze test and examined for hippocampal and cortical pathologies and amyloid plaques using HE,immunohistochemical and thioflavin S staining;ELISA and Evans blue method were used for detecting Aβ_(1-40) and Aβ_(1-42) levels in the brain tissue and serum and assessing blood-brain barrier(BBB)integrity.Immunofluorescence colocalization was used to investigate AQP4 polarization on astrocytes.Western blotting was performed to detect the expressions of VE-cadherin,ZO-1,occludin,β-amyloid precursor protein(APP),BACE1,insulin-degrading enzyme(IDE),LRP1,RAGE,and AQP4 proteins.Results Compared with the control mice,APP/PS1 mice showed significant impairment of learning and memory abilities,increased degeneration or necrosis of hippocampal and cortical neurons,pathological scores,Aβ-positive plaques,elevated Aβ_(1-40) and Aβ_(1-42) levels in the brain tissue and serum,increased BBB permeability,upregulated RAGE expression,lowered expressions of VE-cadherin,LRP1,ZO-1,occludin,and AQP4 proteins,and reduced AQP4-expressing GFAP-positive cells.YGS treatment significantly improved the performance of the transgenic mice in Morris water maze test,reduced hippocampal and cortical pathologies and Aβ-positive plaques,and ameliorated the abnormal changes in Aβ_(1-40) and Aβ_(1-42) levels,BBB permeability,protein expressions of RAGE,VE-cadherin,LRP1,ZO-1,occludin and AQP4,and the number of AQP4-expressing GFAP-positive cells.Conclusion YGS improves learning and memory changes in APP/PS1 mice by ameliorating

关 键 词：异功散 痴呆症 脑水液代谢系统 APP/PS1小鼠 AQP4 

分 类 号：R285.5[医药卫生—中药学]