电针“智三针”对5xFAD小鼠Notch信号通路及突触可塑性的影响  

Effects of electroacupuncture with “Zhi San Zhen” on Notch signaling pathway and synaptic plasticity in 5xFAD mice

作　　者：温华能 林润 王逸潇 王冰水 刘璐 刘传耀 蔡灿鑫 崔韶阳 许明珠 Wen Huaneng;Lin Run;Wang Yixiao;Wang Bingshui;Liu Lu;Liu Chuanyao;Cai Canxin;Cui Shaoyang;Xu Mingzhu(Department of Rehabilitation Medicine,Shenzhen Hospital,Southern Medical University,Shenzhen 518101,Guangdong Province,China;The Third Clinical Medical College of Southern Medical University,Shenzhen 518101,Guangdong Province,China;Deparment of Rehabilitation Medicine,Shenzhen Hospital(Futian),Guangzhou University of Chinese Medicine,Shenzhen 518034,Guangdong Province,China)

机构地区：[1]南方医科大学深圳医院康复医学科,广东省深圳市518101 [2]南方医科大学第三临床医学院,广东省深圳市518101 [3]广州中医药大学深圳医院(福田)康复医学科,广东省深圳市518034

出　　处：《中国组织工程研究》2024年第32期5148-5153,共6页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金青年基金(81904275),项目负责人:许明珠;深圳市科技计划基础研究面上项目(JCY20190801182730737),项目负责人:许明珠;深圳市科技计划基础研究面上项目(JCYJ20190814114207451),项目负责人:王冰水;深圳市福田区卫生公益性科研项目(FTWS2020060),项目负责人:崔韶阳;南方医科大学深圳医院科研基金项目(PT2019GZR08),项目负责人:许明珠。

摘　　要：背景:阿尔茨海默病是以认知功能障碍为主要临床表现的退行性神经系统疾病,针刺是治疗阿尔茨海默病的传统特色疗法,但作用机制尚不明晰。目的:观察电针“智三针”对5xFAD小鼠Notch信号通路、β-淀粉样蛋白及突触可塑性的影响。方法:将16只SPF级、雄性、6月龄5xFAD小鼠随机分为电针“智三针”组和模型组,每组8只,另取8只同条件C57BL/6小鼠作为野生对照组。电针“智三针”组进行电针“智三针”干预,每周5次,连续4周;模型组、野生对照组不进行干预。干预结束后,采用Morris水迷宫初步评价学习记忆能力;硫磺素S染色检测β-淀粉样蛋白斑块沉积情况;Western blot和qPCR检测跨膜受体蛋白Notch 1、Notch 1胞内段(Notch 1 intracellular domain,NICD)、Split多毛增强子1(hairy and enhancer of split 1,Hes 1)、Split多毛增强子5(hairy and enhancer of split 5,Hes 5)、突触生长蛋白(synaptophysin,SYN)、突触后密度蛋白95(postsynaptic density protein-95,PSD-95)及β-淀粉样蛋白的表达水平。结果与结论:①与模型组相比,野生对照组、电针“智三针”组小鼠逃避潜伏期缩短,穿越平台次数及目标象限停留时间增加(P<0.05);②与野生对照组相比,模型组小鼠海马区β-淀粉样蛋白斑块沉积显著增加,而电针“智三针”抑制了β-淀粉样蛋白斑块沉积(P<0.05);③与野生对照组相比,模型组小鼠海马组织中SYN、PSD-95、Notch 1、NICD、Hes 1及Hes 5 mRNA表达降低,β-淀粉样蛋白mRNA表达增加(P<0.05);与模型组相比,电针“智三针”组SYN、PSD-95、Notch 1、NICD、Hes 1及Hes 5 mRNA表达升高,β-淀粉样蛋白mRNA表达降低(P<0.05);④与野生对照组相比,模型组小鼠海马组织中SYN、PSD-95、Notch 1、NICD、Hes 1及Hes 5蛋白表达降低,β-淀粉样蛋白表达增加(P<0.05),与模型组相比,电针“智三针”组SYN、PSD-95、Notch 1、NICD、Hes 1及Hes 5蛋白表达增加,β-淀粉样蛋白表达降低(P<0.05);�BACKGROUND:Alzheimer’s disease is a degenerative neurological disorder characterized primarily by cognitive impairment.Acupuncture is a kind of traditional Chinese medicine therapy for treating Alzheimer’s disease,but its mechanism is not yet clear.OBJECTIVE:To observe the effects of electroacupuncture with“Zhi San Zhen”on the Notch signaling pathway,β-amyloid protein(Aβ)and synaptic plasticity in 5xFAD mice.METHODS:Sixteen male,6-month-old 5xFAD mice,SPF-grade,were randomly divided into the electroacupuncture with“Zhi San Zhen”group(electroacupuncture group)and the model group,with eight mice in each group.Eight SPF-grade,male,6-month-old C57BL/6 mice were used as the wild control(wild)group.The electroacupuncture group received electroacupuncture with“Zhi San Zhen”intervention,5 times a week for 4 consecutive weeks.The model group and the wild group did not receive electroacupuncture intervention.The Morris water maze was used to preliminarily assess their learning and memory abilities.Thioflavin S staining was performed to detect Aβplaque deposition.Western blot and real-time quantitative polymerase chain reaction(RT-qPCR)were used to measure the expression levels of transmembrane receptor protein Notch-1,Notch 1 intracellular domain(NICD),hairy and enhancer of split 1(Hes 1),hairy and enhancer of split 5(Hes 5),synaptophysin(SYN),postsynaptic density protein-95(PSD-95),and Aβ.RESULTS AND CONCLUSION:Compared with the model group,the wild group and the electroacupuncture group showed shortened escape latency,increased platform crossing times,and longer target quadrant dwell time(P<0.05).Compared with the wild group,the model group had significantly increased deposition of Aβplaques,while electroacupuncture with“Zhi San Zhen”inhibited the deposition of Aβplaques in the hippocampus of 5xFAD mice(P<0.05).Compared with the wild group,the model group had decreased mRNA levels of SYN,PSD-95,Notch 1,NICD,Hes 1,and Hes 5 in the hippocampal tissue of mice,and increased mRNA levels of Aβ(P<0.05

关 键 词：阿尔茨海默病 智三针 电针 NOTCH信号通路 5xFAD小鼠 突触可塑性 Β-淀粉样蛋白 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学] R741.05