基于TLR4/NF-κB/NLRP3通路探讨电针对阿尔茨海默病小鼠认知障碍的影响  

Effect of electroacupuncture on cognitive impairment in APP/PS1mice based on TLR4/NF-κB/NLRP3pathway

作　　者：廖冬梅 庞芳 周敏 李怡 杨云昊 郭啸 唐成林[1] LIAO Dong-mei;PANG Fang;ZHOU Min;LI Yi;YANG Yun-hao;GUO Xiao;TANG Cheng-lin(College of Traditional Chinese Medicine,Chongqing Medical University,Chongqing 400016,China)

机构地区：[1]重庆医科大学中医药学院,重庆400016

出　　处：《针刺研究》2022年第7期565-572,共8页Acupuncture Research

基　　金：国家自然科学基金项目(No.81273870);重庆市科卫联合中医药科技项目(No.ZY201801007);重庆市渝中区科技计划项目(No.20200116);重庆市教委青年项目(No.KJQN201900)。

摘　　要：目的:观察电针对APP/PS1双转基因小鼠结肠和海马中紧密连接相关蛋白Claudin-5、ZO-1及海马Toll样受体4/核转录因子κB/NOD样受体蛋白3(TLR4/NF-κB/NLRP3)通路的影响,探讨电针改善阿尔茨海默病认知障碍的机制。方法:5月龄雄性APP/PS1双转基因小鼠随机分为模型组、电针组,同月龄雄性C57BL/6小鼠作为对照组,每组9只。电针组电针“百会”“大肠俞”“足三里”,15min/次,每周5次,干预5周。干预结束后,用Morris水迷宫评估小鼠学习记忆能力,尼氏染色法观察小鼠海马区病理形态,免疫组织化学法检测小鼠脑组织β淀粉样蛋白(Aβ)表达,ELISA法检测小鼠结肠、血清、海马组织中脂多糖(LPS)含量,Western blot法检测小鼠结肠Claudin-5、ZO-1和海马Claudin-5、ZO-1及TLR4/NF-κB/NLRP3通路相关蛋白的表达。结果:与对照组比较,从第3天开始,模型组小鼠逃避潜伏期延长(P<0.05,P<0.01),穿越原平台次数、目标象限停留时间百分比显著减少(P<0.01),结肠、血清及海马组织中LPS含量显著升高(P<0.01),海马TLR4、NF-κB p65、NLRP3、Caspase-1、白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)蛋白表达及脑组织Aβ表达均显著升高(P<0.01),结肠及海马Claudin-5、ZO-1蛋白表达均显著降低(P<0.01)。与模型组比较,电针组小鼠第4、5天逃避潜伏期缩短(P<0.05,P<0.01),穿越原平台次数、目标象限停留时间百分比增加(P<0.01,P<0.05),血清及海马组织LPS含量降低(P<0.05),海马TLR4、NF-κB p65、NLRP3、Caspase-1、IL-1β、TNF-α蛋白表达及脑组织Aβ表达均降低(P<0.05,P<0.01),结肠及海马Claudin-5、ZO-1蛋白表达升高(P<0.05,P<0.01)。对照组小鼠海马CA1和CA3区神经元排列整齐、紧密;模型组神经元排列疏散,层数减少,部分细胞固缩、深染,尼氏小体数量减少;电针组神经元排列相对紧密,层数增加,无明显固缩,尼氏小体数量增加。结论:电针可能通过上调Claudin-5、ZO-1的表达,减�Objective To observe the effect of electroacupuncture(EA)on the expressions of tight junction related proteins Claudin-5,ZO-1in the colon and hippocampus,Toll-like receptor 4/nuclear factor-kappa B/NOD-like receptor protein 3(TLR4/NF-κB/NLRP3)pathway in the hippocampus of APP/PS1mice,so as to explore its mechanisms underlying improvement of cognitive impairment.Methods Eighteen 5-month-old male APP/PS1mice were equally randomized into model and EA groups,and nine 5-month-old male C57BL/6mice were used as the normal control.EA(2Hz,1mA)was applied to“Baihui”(GV20),“Dachangshu”(BL25)and“Zusanli”(ST36)for 15min,once daily,5days a week for 5weeks.The Morris water maze swimming test was used to evaluate the mice’s cognitive impairment.Nissl staining was used to observe the pathological morphology of hippocampus.The expression of amyloidβ-peptide(Aβ)in brain tissue was detect by immunohistochemistry;the contents of lipopolysaccharide(LPS)in colon,serum and hippocampus were detected by ELISA;the expression levels of Claudin-5,ZO-1in colon and hippocampus,and TLR4/NF-κB/NLRP3pathway related proteins in hippocampus were detected by Western blot.Results Compared with the normal group,the escape latency of the mice in the model group was prolonged from the 3^(rd) day(P<0.05,P<0.01),the number of crossing the platform and the percentage of target quadrant residence time were significantly decreased(P<0.01),and the contents of LPS in colon,serum and hippocampus were significantly increased(P<0.01),the expression levels of TLR4,NF-κB p65,NLRP3,Caspase-1,interleukin(IL)-1βand tumor necrosis factor(TNF)-αin hippocampus and Aβin brain tissue were significantly increased(P<0.01),while the expression levels of Claudin-5,ZO-1in colon and hippocampus were significantly decreased(P<0.01).Compared with the model group,the escape latency of mice in the EA group was shortened from the 4^(th) day(P<0.05,P<0.01),the number of crossing the platform and the percentage of target quadrant residence time were increased(P<0.

关 键 词：阿尔茨海默病 脑-肠轴 电针 TLR4/NF-κB/NLRP3通路 认知障碍 

分 类 号：R245.97[医药卫生—针灸推拿学]