富马酸二甲酯对亚砷酸钠诱发小鼠胰岛MIN6细胞氧化应激损伤的保护作用  

Protective role of dimethyl fumarate against sodium arsenite-induced oxidative stress injury in MIN6 cells of pancreatic islet

作　　者：赵紫汐 王司琪 李雪研 徐春雪 赵曦 杨蓓[1] Zhao Zixi;Wang Siqi;Li Xueyan;Xu Chunxue;Zhao Xi;Yang Bei(Department of Histology and Embryology,College of Basic Medical Sciences,China Medical University,Shenyang 110122,China;Clinical Medicine Major of Grade 2019,China Medical University,Shenyang 110122,China;Basic Medicine Major of Grade 2017,China Medical University,Shenyang 110122,China)

机构地区：[1]中国医科大学基础医学院组织学与胚胎学教研室,沈阳110122 [2]中国医科大学,沈阳110122

出　　处：《解剖学杂志》2023年第2期120-125,共6页Chinese Journal of Anatomy

基　　金：国家自然科学基金(82171882);辽宁省教育厅资助项目(JCZR2020002);2021年辽宁省大学生创新创业训练计划资助项目(S202110159043)。

摘　　要：目的:探讨富马酸二甲酯(DMF)对亚砷酸钠(NaAsO2)诱发小鼠胰岛MIN6细胞氧化应激损伤的保护作用及相关机制。方法:筛选DMF最适处理浓度,将MIN6细胞随机分为正常对照组、NaAsO2组、低剂量DMF预处理+NaAsO2组、高剂量DMF预处理+NaAsO2组。MTT法检测细胞活性;TUNEL荧光染色法检测细胞凋亡;DCFH-DA荧光探针法检测活性氧自由基(ROS)水平;免疫细胞荧光法观察细胞内核因子E2相关因子2(Nrf2)定位表达;免疫印迹检测Nrf2蛋白表达水平;Real-time RT-PCR检测Nrf2、血红素加氧酶1(Ho-1)、硫氧还蛋白1(Trx1)、γ-谷氨酰半胱氨酸连接酶催化亚单位(Gclc)、γ-谷氨酰半胱氨酸连接酶调节亚单位(Gclm)mRNA表达水平。结果:与NaAsO2组比较,DMF预处理可以显著提高MIN6细胞活性,减少细胞凋亡数量,降低细胞内ROS的水平。DMF预处理+NaAsO2组MIN6细胞内的Nrf2 mRNA和蛋白表达明显高于NaAsO2组,特别是细胞核内Nrf2表达显著增多,且呈显著的剂量依赖性。DMF预处理+NaAsO2组MIN6细胞Ho-1、Trx1、Gclc和Gclm mRNA表达亦出现明显增加。结论:DMF可能通过上调Nrf2/ARE信号通路,拮抗亚砷酸钠对胰岛MIN6细胞氧化应激损伤,发挥保护作用。Objective:To investigate the protective role of dimethyl fumarate(DMF)against sodium arsenite(NaAsO2)-induced oxidative stress injury in MIN6 cells of pancreatic islet and related mechanisms.Methods:MIN6 cells were randomly divided into four groups:the control group,NaAsO2 group,low-dose DMF pretreatment+NaAsO2 group,and high-dose DMF pretreatment+NaAsO2 group according to the screening of DMF optimal treatment concentration.Cell viability was measured by MTT.Apoptosis was detected by TUNEL fluorescence staining.Reactive oxygen species(ROS)was measured by DCFH-DA fluorescence probe.Localization of nuclear factorerythroid 2-related factor 2(Nrf2)was observed by immunocytofluorescence.Nrf2 protein expression was examined by Western blotting.Real-time RT-PCR was performed to detect mRNA expressions of Nrf2,Heme oxygenase 1(Ho-1),Thioredoxin 1(Trx1),Catalytic subunit of glutamate cysteine ligase(Gclc),and modifier subunit of glutamate cysteine ligase(Gclm).Results:In comparison with the NaAsO2 group,the DMF pretreatment could dramatically increase the cell viability,decrease the number of apoptotic cells and the level of intracellular ROS in MIN6 cells.The mRNA and protein expressions of Nrf2 in the DMF pretreatment+NaAsO2 group was markedly higher than that in the NaAsO2 group,especially the expression of Nrf2 in the nucleus was notably enhanced in a dose-dependent fashion.In addition,mRNA expressions of Ho-1,Trx1,Gclc,and Gclm also appeared to be statistically augmented in the DMF pretreatment+NaAsO2 group.Conclusion:DMF might exert protective roles by upregulating the Nrf2/ARE signaling pathway to antagonize the oxidative stress injury induced by sodium arsenite in MIN6 cells of pancreatic islet.

关 键 词：富马酸二甲酯 亚砷酸钠 核因子E2相关因子2 氧化应激 MIN6细胞 

分 类 号：R587.1[医药卫生—内分泌]