胞外信号调节激酶信号通路对亚砷酸钠诱导的小鼠海马神经元细胞凋亡的调控作用  

Role of the extracellular signal-regulated kinase pathway in arsenic-induced apoptosis in mouse hippocampal neuron cells

作　　者：陈峣 于漫[1] 李昕[2] 尉玉婷 刘旭丹[2] 王欢欢 王梅[1] CHEN Yao;YU Man;LI Xin;WEI Yuting;LIU Xudan;WANG Huanhuan;WANG Mei(Department of Public Health and Preventive Medicine,School of Integrated Traditional and Western Medicine,Liaoning University of Traditional Chinese Medicine,Shenyang 110847,China;Department of Occupational Health,School of Public Health,China Medical University,Shenyang 110122,China)

机构地区：[1]辽宁中医药大学中西医结合学院公共卫生与预防医学教研室,沈阳110847 [2]中国医科大学公共卫生学院劳动卫生教研室,沈阳110122

出　　处：《中国医科大学学报》2025年第1期24-29,共6页Journal of China Medical University

基　　金：辽宁省科学技术计划(2022-MS-285)。

摘　　要：目的探讨活性氧(ROS)及其介导的胞外信号调节激酶(ERK)/cAMP反应元件结合蛋白(CREB)信号通路在亚砷酸钠(NaAsO_(2))诱导的小鼠海马神经元HT-22细胞凋亡中的作用机制。方法体外培养NaAsO_(2)暴露HT-22细胞,建立体外NaAsO_(2)暴露模型。将细胞分为对照组,4、6、8、10μmol/L NaAsO_(2)暴露组,N-乙酰半胱氨酸(NAC,5 mmol/L)独立干预组,NAC(5 mmol/L)与NaAsO_(2)(10μmol/L)联合干预组。二氯荧光素检测ROS水平;Western blotting检测ERK/CREB信号通路相关蛋白和凋亡相关蛋白水平;流式细胞术检测细胞凋亡率。结果与对照组相比,4、6、8、10μmol/L NaAsO_(2)暴露组细胞中ROS水平显著升高,细胞核中p-ERK蛋白水平显著降低(P<0.05),细胞中p-CREB、Bcl-2蛋白水平显著降低(P<0.05)。与对照组相比,8、10μmol/L NaAsO_(2)暴露组细胞凋亡率显著升高(P<0.05)。与10μmol/L NaAsO_(2)暴露组相比,NAC与NaAsO_(2)联合干预组细胞中ROS水平显著降低,细胞核中p-ERK蛋白水平显著升高(P<0.05),细胞中p-CREB、Bcl-2蛋白水平显著升高,细胞凋亡率显著降低(P<0.05)。结论NaAsO_(2)暴露激活细胞氧化应激反应,通过阻碍p-ERK核易位过程抑制CREB信号通路,从而诱发细胞凋亡。Objective To investigate the role of reactive oxygen species(ROS)and the extracellular signal-regulated kinase(ERK)/cAMP response element binding protein(CREB)signaling pathway in arsenic-induced apoptosis in HT-22 cells.Methods HT-22 cells were cultured in vitro and exposed to NaAsO_(2).The cells were divided into the following groups:control group,4,6,8,and 10μmol/L NaAsO_(2) groups,NAC(5 mmol/L)group,and NAC(5 mmol/L)+NaAsO_(2)(10μmol/L)group.ROS levels were measured using a DCFH-DA assay.The expression of ERK/CREB signaling pathway-related and apoptosis-related proteins were analyzed by Western blotting.Apoptosis rates were evaluated using flow cytometry.Results Compared with the control group,ROS levels in HT-22 cells significantly increased,while p-ERK protein level in the nucleus and p-CREB and Bcl-2 protein levels in the cells significantly decreased in the NaAsO_(2) only exposed groups(P<0.05).Additionally,the apoptosis rate significantly increased in the 8 and 10μmol/L NaAsO_(2) groups compared with the control group(P<0.05).In the NAC+NaAsO_(2) group,ROS levels and the apoptosis rate significantly decreased while p-ERK protein level in the nucleus and p-CREB and Bcl-2 protein levels in the cells significantly increased compared with the 10μmol/L NaAsO_(2) group(P<0.05).Conclusion Arsenic exposure induces oxidative stress,inhibits the nuclear translocation of p-ERK,and disrupts the CREB signaling pathway,leading to apoptosis.

关 键 词：砷 胞外信号调节激酶 CAMP反应元件结合蛋白 凋亡 神经损伤 

分 类 号：R994.6[医药卫生—毒理学]