亚慢性铝暴露致大鼠认知障碍ALKBH5/PTEN/AKT信号通路机制  

Sub-chronic aluminum exposure induces cognitive impairment through ALKBH5/PTEN/AKT signaling pathway in rats

作　　者：刘衍 张晓宇 张伟涛 萧非凡 崔平 王彬鸿 陈薪茹 姜斌 陈欢 林立[1,2] 张璟 李欢[1,2] LIU Yan;ZHANG Xiaoyu;ZHANG Weitao;XIAO Feifan;CUI Ping;WANG Binhong;CHEN Xinru;JIANG Bin;CHEN Huan;LIN Li;ZHANG Jing;LI Huan(School of Public Health,Jining Medical University,Jining,Shandong 272000,China;不详)

机构地区：[1]济宁医学院公共卫生学院,山东济宁272000 [2]滨州医学院公共卫生学院,山东滨州256600 [3]山东第一医科大学(山东省医学科学院)公共卫生与健康管理学院,山东济南250117 [4]嘉祥县疾病预防控制中心,山东嘉祥272400

出　　处：《中国职业医学》2024年第2期144-149,155,共7页China Occupational Medicine

基　　金：山东省自然科学基金青年项目(ZR2022QH114);贺林院士新医学临床转化工作站科研基金(JYHL2022MS07);山东省医药卫生科技发展计划(2019WSB33008);济宁医学院博士科研启动基金(600941001);济宁医学院高层次科研培育计划项目(JYGC2023KJ004)。

摘　　要：目的探讨麦芽酚铝暴露对miR-193a-3p、去甲基化酶AlkB同源蛋白5(ALKBH5)、第10号染色体缺失的磷酸酶和张力蛋白同源物基因(PTEN)和蛋白激酶B(AKT)的影响,以及miR-193a-3p是否通过调控ALKBH5/PTEN/AKT信号通路参与铝致认知障碍的机制。方法将无特定病原体级健康雄性SD大鼠随机分为对照组和低、中、高剂量组,每组8只。3个剂量组大鼠分别予浓度为10.00、20.00和40.00μmol/kg体质量的麦芽酚铝溶液腹腔注射染毒,对照组大鼠予等体积0.9%氯化钠溶液;每周连续染毒5 d,持续3个月。染毒结束后,以新物体识别实验检测大鼠学习记忆能力,以实时荧光定量聚合酶链反应法检测大鼠海马组织中miR-193a-3p和B淋巴细胞瘤-2(Bcl-2)、Bcl-2相关X蛋白(Bax)、半胱氨酸天冬氨酸蛋白酶-3(Caspase-3)的mRNA相对表达水平,以蛋白质印迹法检测大鼠海马组织中ALKBH5、PTEN和AKT2蛋白的表达。结果高剂量组大鼠辨别指数和偏好指数分别低于对照组和低剂量组(P值均<0.05)。高剂量组大鼠海马组织中miR-193a-3p和Bcl-2 mRNA相对表达水平分别低于对照组和低剂量组(P值均<0.05),Bax mRNA相对表达水平分别高于对照组和低剂量组(P值均<0.05),Caspase-3 mRNA相对表达水平分别高于其他3组(P值均<0.05)。高剂量组大鼠海马组织中ALKBH5蛋白相对表达水平低于对照组(P<0.05),PTEN蛋白相对表达水高于对照组和低剂量组(P值均<0.05),AKT2蛋白相对表达水平低于对照组和低剂量组(P值均<0.05)。结论亚慢性铝暴露可抑制大鼠海马组织中miR-193a-3p的表达,从而破坏ALKBH5/PTEN/AKT通路,影响正常的神经元稳态和细胞功能;此机制可能在铝诱导的认知障碍过程中发挥重要作用。Objective To investigate the effects of maltol aluminum exposure on miR-193a-3p,demethylase AlkB homolog 5(ALKBH5),phosphatase and tensin homolog deleted on chromosome ten(PTEN)and protein kinase B(AKT),and whether miR-193a-3p is involved in aluminum-induced cognitive impairment by regulating ALKBH5/PTEN/AKT signaling pathway.Methods Specific pathogen-free male SD rats were randomly divided into control group and low-,medium-and high-dose groups according to their body weight,with eight rats in each group.Rats in the low-,medium-,and high-dose groups were intraperitoneally injected with maltol aluminum solution at concentrations of 10.00,20.00,and 40.00μmol/kg body weight,respectively,while the rats in control group were given an equal volume of 0.9%sodium chloride solution.Rats were injected for five days every week for three months.After injection,the novel object recognized test was used to assess the learning and memory ability of the rats.The relative expression of miR-193a-3p and B-cell lymphocytoma-2(Bcl-2),Bcl-2 associated X protein(Bax)and cysteine aspartate protease-3(Caspase-3)mRNA in rat hippocampus was detected using the real-time quantitative polymerase chain reaction.The relative protein expression of ALKBH5,PTEN,and AKT2 in the rat hippocampus was detected using Western blot.Results The discrimination index and the preference index of the new object recognition test of the rats in high-dose group were lower than those in control group and low-dose group(all P<0.05).The relative expression of miR-193a-3p and Bcl-2 mRNA in the hippocampus of the rats in high-dose group was lower than those in control group and lowdose group(all P<0.05).The relative mRNA expression of Bax in the high-dose group was higher than those in the control group and low-dose group(both P<0.05).The relative mRNA expression of Caspase-3 of the rats in the high-dose group was higher than that in the other three groups(both P<0.05).The relative protein expression of ALKBH5 in the hippocampus of the rats in the high-dose group wa

关 键 词：铝 miR-193a-3p 去甲基化酶AlkB同源蛋白5 张力蛋白同源基因 蛋白激酶B 认知障碍 信号通路 大鼠 

分 类 号：R135.1[医药卫生—劳动卫生]