Nrf2 /ARE 信号通路对快速老化小鼠认知功能和神经炎症的影响  

Effect of the Nrf2 / ARE signaling pathway on cognitive function and neuroinflammation in senescence accelerated mice

作　　者：徐玉珠 步玮[2] 杨玲[1] 王锦阳[1] 任慧玲[1] XU Yuzhu;BU Wei;YANG Ling(Department of Neurology,the Third Hospital of Hebei Medical University,Hebei,Shijiazhuang 050017,China;不详)

机构地区：[1]河北医科大学第三医院神经内科,石家庄市050017 [2]河北医科大学第三医院神经外科,石家庄市050017

出　　处：《河北医药》2024年第21期3205-3211,共7页Hebei Medical Journal

基　　金：河北省医学科学研究课题计划(编号:20190607)。

摘　　要：目的探讨核因子E2相关因子2(Nrf2)/抗氧化反应元件(ARE)信号通路对快速老化小鼠(SAMP8)认知功能和神经炎症的影响。方法选用4个月大的雄性抗快速老化系小鼠1(SAMR1)及SAMP8小鼠,颅脑海马区域注射慢病毒介导的Nrf2-ShRNA或对照的GFP,靶向性下调目的基因Nrf2在小鼠体内的表达。小鼠随机分为4组:R1+GFP组、R1+Nrf2 ShRNA组、P8+GFP组、P8+Nrf2 ShRNA组,每组10只。新异物体识别实验、水迷宫实验和避暗实验等检测小鼠的认知能力,免疫组织荧光染色法检测微管结合蛋白2(MAP2)和目的基因Nrf2在小鼠海马区的表达水平,蛋白免疫印迹(Western blot)法检测4组小鼠海马组织中Nrf2、白介素-6(IL-6)以及肿瘤坏死因子-α(TNF-α)水平。结果新异物体识别实验中,9个月龄快速老化SAMP8实验小鼠与同月龄的SAMR1小鼠相比,在分辨新物体的能力方面表现较差(P<0.05),而2组SAMP8小鼠相比,在瞬时的1 h测试时,2组小鼠对新物体的认知指数差异无统计学意义(P>0.05),然而在进行延迟记忆的24 h的时间测试时,P8+Nrf2 ShRNA组小鼠的认知指数出现明显下降(P<0.05)。在Morris水迷宫测试中,SAMP8组小鼠与同一年龄段的SAMR1组小鼠相比,逃避潜伏期延长(P<0.05),而2组SAMR1小鼠在逃避潜伏期差异无统计学意义(P>0.05),老化的SAMP8小鼠在空间探索实验中的穿越平台次数较SAMR1组明显降低,其中,P8+Nrf2 ShRNA组的小鼠穿越平台次数下降更为严重(P<0.05),而2组SAMR1组的小鼠相比,其穿越平台次数并无显著变化(P>0.05)。在避暗实验中,9月龄的SAMP8组小鼠比同月龄的SAMR1组小鼠电击次数增加,P8+Nrf2 ShRNA组小鼠的电击次数比P8+GFP组增加更明显,且此组小鼠的逃避潜伏时呈中等程度下降趋势(P<0.05)。在免疫组织荧光染色中,SAMP8小鼠的海马区MAP2阳性面积比SAMR1小鼠显著降低(P<0.05),相比于对照的P8+GFP组小鼠,试验组的P8+Nrf2 ShRNA小鼠在海马CA3区的MAP2表达强度和阳Objective To explore the impact of the nuclear factor E2-related factor 2(Nrf2)/antioxidant response element(ARE)signaling pathway on cognitive function and neuroinflammation in the Senescence Accelerated Mouse-Prone 8(SAMP8)mice.Methods Male Senescence Accelerated Mouse Resistant 1(SAMR1)and SAMP8 mice with 4 months old were selected.SAMR1 or SAMP8 mice were injected with lentivirus-mediated Nrf2-ShRNA in the hippocampus to down-regulate Nrf2,or negative control(GFP),with 10 mice per group.The cognitive ability of mice was tested by novel object recognition test,water maze experiment and light-dark box test.Positive expressions of microtubule-associated protein 2(MAP2)and target gene Nrf2 in the hippocampus were observed by immunofluorescence staining.Western blot was used to examine protein levels of Nrf2,interleukin 6(IL-6),and tumour necrosis factor alpha(TNF-α)in the hippocampus.Results The novel object recognition experiment data showed a worse ability to discriminate novel objects in 9-month-old SAMP8 mice than age-matched SAMR1 mice(P<0.05).No significant difference in the 1-h recognition index between SAMP8 mice transfected with Nrf2-ShRNA or GFP(P>0.05).Recognition index at 24 h of delayed memory was significantly lower in SAMP8 mice transfected with Nrf2-ShRNA than those transfected with GFP(P<0.05).The results of the Morris water maze test showed that the escape latency was significantly longer in SAMP8 mice than age-matched SAMR1 mice(P<0.05),while no significant difference in the escape latency was detected in SAMR1 mice transfected with Nrf2-ShRNA or GFP(P>0.05).In the spatial exploration experiment,the number of traveling across the platform was significantly less in SAMP8 mice than age-matched SAMR1 mice,which was much less in SAMP8 mice transfected with Nrf2-ShRNA than those transfected with GFP(P<0.05),while no significant difference in the number of traveling across the platform was detected in SAMR1 mice transfected with Nrf2-ShRNA or GFP(P>0.05).In the light-dark box test,9-month SAMP8 mice

关 键 词：Nrf2/ARE信号通路 阿尔茨海默病 炎性因子 认知功能 

分 类 号：R741.02[医药卫生—神经病学与精神病学]