CREB与miR-132-3p在铝致大鼠海马tau蛋白异常磷酸化过程中的作用  

Effects of CREB and miR-132-3p on aluminum-induced abnormal phosphorylation of tau protein in rat hippocampus

作　　者：肖雯洁 徐旭 雷敏敏 杨晓娟[1,2] XIAO Wenjie;XU Xu;LEI Minmin;YANG Xiaojuan(School of Public Health,Shanxi Medical University,Taiyuan,Shanxi 030001,China;Department of Geriatrics,First Hospital of Shanxi Medical University,Taiyuan,Shanxi 030001,China)

机构地区：[1]山西医科大学公共卫生学院,山西太原030001 [2]山西医科大学第一医院老年科,山西太原030001

出　　处：《环境与职业医学》2024年第11期1270-1276,共7页Journal of Environmental and Occupational Medicine

基　　金：山西省留学人员科技活动择优资助项目(20200008);山西省基础研究计划项目(202103021224398);山西省回国留学人员科研资助项目(2023-188)。

摘　　要：[背景]环磷酸腺苷反应元件结合蛋白(CREB)与miR-132-3p已被证实与认知功能障碍有关。课题组前期已证明铝的神经毒性与tau蛋白异常磷酸化有关,但关于铝是否通过CREB与miR-132-3p影响tau蛋白异常磷酸化的研究尚未报道。[目的]探讨铝致大鼠海马tau蛋白异常磷酸化过程中CREB与miR-132-3p的变化情况。[方法]随机将28只体重相近的2月龄SD大鼠均匀划分为4个组,包括对照组(使用生理盐水)和低、中、高剂量的铝染毒组[使用10、20、40μmol·kg^(-1)麦芽酚铝Al(mal)3]。大鼠隔天称重染毒,染毒周期为3个月。染毒后通过水迷宫行为学实验反应大鼠学习记忆能力的变化情况;运用实时荧光定量聚合酶链式反应(qRT-PCR)技术来测定miR-132-3p基因的表达水平;使用蛋白免疫印记(WB)技术来测定CREB、磷酸化CREB(p-CREB)(Ser133)、RAS GTP酶激活蛋白1(RASA1)、tau和p-tau(Ser396)蛋白的表达水平。[结果]Morris水迷宫行为学实验结果:在定位航行实验阶段(第1天至第5天)记录实验动物的逃避潜伏期时长,3种剂量的铝染毒组大鼠的逃避潜伏期较于对照组大鼠均有延长(P<0.05);在第6天进行的空间探索实验中,与对照组和低剂量铝染毒组的大鼠相比,中、高剂量铝染毒组大鼠在目标象限的停留时间和穿越平台的次数均减少(P<0.05),且高剂量铝染毒组的大鼠在目标象限的停留时间比中剂量铝染毒组短(P<0.05)。qRT-PCR结果:随着Al(mal)3剂量的升高,大鼠海马miR-132-3p基因表达水平递减(P<0.05)。WB结果显示,与对照组及低剂量铝染毒组相比,中、高剂量铝染毒组大鼠海马的CREB和p-CREB(Ser133)蛋白表达水平均降低(P<0.05),同时低剂量铝染毒组的p-CREB(Ser133)蛋白表达水平也低于对照组(P<0.05),经方差分析趋势检验和Pearson相关分析后发现铝染毒剂量的进一步升高将导致CREB、p-CREB(Ser133)蛋白表达水平的进一步降低(F=36.429,P<0.001;F=78.672,P<0.001),铝�[Background]Cyclic AMP response element binding protein(CREB)and miR-132-3p have been proved to be related to many neurodegenerative diseases.Our research group previously has demostrated that the neurotoxicity of aluminum is relevant to abnormal phosphorylation of tau protein,but whether aluminum affects the abnormal phosphorylation of tau protein through GREB and miR-132-3p has not been reported yet.[Objective]To investigate the effect of aluminum on CREB and miR-132-3p during abnormal phosphorylation of tau protein in rat hippocampus.[Methods]Twenty-eight two-month-old SD rats with comparable weigh,were randomly assigned to four groups:control group(saline)and low,middle,and high dose exposure groups[10,20,and 40μmol·kg^(−1) Al(mal)3]with each group containing 7 rats,and the exposureperiod was 3 months by intraperitoneal injection every other day.After rats’exposure to aluminum,Morris water maze was employedto assess their capabilities of learning and memory.The miR-132-3p gene expression level was detected by quantitative real-time PCR(qRT-PCR).The levels of CREB,phosphorylated CREB(p-CREB)(Ser133),RAS p21 protein activator 1(RASA1)tau,and p-tau(Ser396)proteinswere determined by Western blot.[Results]The results of Morris water maze showed that in the navigation experiment(from first day to the fifth day),the average escapelatency of the rats exposed to three doses of aluminum was longer than that of the control rats(P<0.05).The middle dose group and thehigh dose group demonstrated shorter duration and lower frequency of platform traversal in the designated quadrant when comparedto the control group and the low dose group(P<0.05).Moreover,the duration in the target quadrant of the rats exposed to high dosealuminum was shorter than that of the rats exposed to medium dose aluminum(P<0.05).The results of Morris water maze suggestedthat aluminum could damage the learning and memory ability of rats.The qRT-PCR findings indicated a decline in miR-132-3p gene expressionin rat hippocampus correlating with highe

关 键 词：铝 miR-132-3p 环磷酸腺苷反应元件结合蛋白 TAU蛋白磷酸化 学习记忆 

分 类 号：R12[医药卫生—环境卫生学]