补肾活血针刺法对SAMP8小鼠认知功能及海马神经元沉默信息调节因子2调控作用的研究  

Effect of Bushen Huoxue acupuncture on cognitive function and silent information regulator 2 in hippocampal neurons of SAMP8 mice

作　　者：杨聘[1] 徐融[1] 谢立全 YANG Pin;XU Rong;XIE Liquan(Department of Geriatrics,Hangzhou Hospital of Traditional Chinese Medicine,Hangzhou,Zhejiang 310007,China)

机构地区：[1]杭州市中医院老年病科,杭州310007

出　　处：《浙江中西医结合杂志》2025年第1期6-12,共7页Zhejiang Journal of Integrated Traditional Chinese and Western Medicine

基　　金：浙江省中医药科技计划项目(No.2023ZL536)。

摘　　要：目的探究补肾活血针刺法对加速衰老的小鼠模型SAMP8小鼠认知功能和海马神经元沉默信息调节因子2(SIRT2)介导的内质网蛋白4B(RTN4B)/β淀粉样前体蛋白裂解酶1(BACE1)通路的影响。方法6只SAMR1小鼠为正常对照组,18只SAMP8小鼠随机分为模型对照组、针刺治疗组、非针刺治疗组,各6只。针刺治疗组血海、膈俞施捻转泻法,肾俞、百会施捻转补法,运针1 min,留针10 min;非针刺治疗组在非经非穴点进行抓捉刺激;均干预8周。通过Morris水迷宫试验评估小鼠认知功能。采用免疫组织化学染色和酶联免疫吸附试验评估海马神经元小胶质细胞的激活和β淀粉样蛋白(Aβ)沉积。通过蛋白质免疫印迹检测SIRT2途径相关蛋白SIRT2、RTN4B、BACE1和β淀粉样前体蛋白C-末端片段(APP-CTF)的表达水平。结果与正常对照组比较,模型对照组逃避潜伏期延长,目标象限停留时间减少,穿越平台次数减少(P<0.05),海马组织Aβ42和CD68阳性面积增大(P<0.01),血清Aβ42水平增高,海马组织中SIRT2、BACE1和APP-CTF表达明显增加,RTN4B表达降低(P<0.05)。与模型对照组及非针刺治疗组比较,针刺治疗组逃避潜伏期缩短[第1天:(37.80±10.42)s比(49.80±6.14)s、(44.60±7.40)s,P<0.05;第2天:(36.80±12.69)s比(48.80±5.97)s、(44.20±7.72)s,P<0.05;第3天:(38.60±9.71)s比(51.20±5.54)s、(43.60±6.46)s,P<0.05;第4天:(36.00±11.20)s比(46.40±5.81)s、(45.20±7.36)s,P<0.05;第5天:(36.60±11.37)s比(47.80±5.31)s、(43.80±9.44)s,P<0.05],目标象限停留时间增加[(7.83±0.98)s比(1.00±0.63)s、(3.33±0.52)s,P<0.05],穿越平台次数增加[(13.33±1.03)次比(3.17±1.17)次、(7.33±0.52)次,P<0.05];海马组织Aβ42和CD68阳性面积减少(P<0.01),血清Aβ42水平显著降低[(11.38±1.57)μg/mL比(23.14±2.41)μg/mL、(17.16±1.27)μg/mL,P<0.05];海马组织中SIRT2[(1.98±0.19)比(4.21±0.31)、(3.22±0.23),P<0.05或P<0.01]、BACE1[(1.81±0.14)比(2.80±0.19)、(2.43±0.13),P<0.05或P<0.01]和APP-CObjective To investigate the effects of Bushen Huoxue acupuncture on cognitive function and the silent information regulator 2(SIRT2)-mediated reticulon 4B(RTN4B)/beta-site amyloid precursor protein cleaving enzyme 1(BACE1)pathway in hippocampal neurons of senescence-accelerated mouse prone 8(SAMP8)mice.Methods Six SAMR1 mice served as a normal control group,while 18 SAMP8 mice were randomly divided into three groups:a model control group,an acupuncture treatment group,and a non-acupuncture treatment group,with six mice in each group.The acupuncture treatment group received twisting and purging technique at Xuehai and Geshu,and twisting and tonifying technique at Shenshu and Baihui,with needle manipulation for 1 minute and retention for 10 minutes.The non-acupuncture treatment group underwent grasping stimulation at non-meridian and non-acupoint sites.All interventions lasted for 8 weeks.Cognitive function was evaluated using the Morris water maze test.Immunohistochemistry and enzyme-linked immunosorbent assay were used to assess the activation of hip-pocampal neuron and beta-amyloid protein(Aβ)deposition.Western blotting was used to examine the expression levels of SIRT2 pathway-related proteins,including SIRT2,RTN4B,BACE1,and beta-amyloid precursor protein C-terminal fragment(APP-CTF).Results Compared with the normal control group,the model control group showed prolonged escape latency,reduced time spent in the target quadrant,and decreased platform crossing times(P<0.05).Additionally,the model control group exhibited increased Aβ42 and CD68-positive areas in hippocampal tissue(P<0.01),elevated serum Aβ 42 levels,significantly upregulated expression of SIRT2,BACE1,and APP-CTF,and downregulated expression of RTN4B(P<0.05).Compared with the model control group and the non-acupuncture treatment group,the acupuncture treatment group showed shortened escape latency on day 1 to day 5[day 1:(37.80±10.42)s vs.(49.80±6.14)s,(44.60±7.40)s,P<0.05;day 2:(36.80±12.69)s vs.(48.80±5.97)s,(44.20±7.72)s,P<0.05;day 3:(3

关 键 词：小鼠 补肾活血针刺法 AΒ沉积 认知功能 沉默信息调节因子2 内质网蛋白4B/β淀粉样前体蛋白裂解酶1 

分 类 号：R24[医药卫生—中医临床基础]