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作 者:宋晨晨 王威 尧青[2] 莫启贵 刘爱梅 SONG Chen-chen;WANG Wei;LIU Ai-mei(School of Pharmacy,Xianning Medical College,Hubei University of Science and Technology,Xianning Hubei 437100,China)
机构地区:[1]湖北科技学院医学部药学院,湖北咸宁437100 [2]湖北科技学院医学部医药研究院
出 处:《湖北科技学院学报(医学版)》2024年第5期414-417,共4页Journal of Hubei University of Science and Technology(Medical Sciences)
基 金:湖北科技学院2022年度医学科研专项(2022YKY17);湖北科技学院博士启动基金项目(BK202315);湖北省青年基金(2023AFB537)。
摘 要:目的探讨铁死亡在呕吐毒素(DON)诱导的L02细胞毒性中的作用。方法用1640培养基培养人正常肝细胞L02,用不同浓度的DON以及铁死亡抑制剂Fer-1处理细胞。荧光定量检测DON对铁死亡相关基因的表达变化,C11 BODIPY581/591荧光探针检测DON对细胞脂质ROS水平的影响,Ferro Orange荧光探针检测DON对细胞内Fe^(2+)水平的影响,CCK-8法检测Fer-1对DON暴露下细胞活力的影响。结果DON抑制铁死亡相关基因的表达,尤其是谷胱甘肽过氧化物酶4(GPX4)、溶质载体家族7成员11(SLC7A11),促进转铁蛋白受体1(TFR1)的表达。增加细胞脂质活性氧(ROS)和细胞内Fe^(2+)水平,诱导铁死亡,使细胞活力降低。Fer-1的预处理抑制了DON诱导的细胞铁死亡,提高了细胞的活力。结论DON可导致L02细胞铁死亡,该过程与SLC7A11/GPX4通路抑制有关。Objective To investigate the role of ferroptosis in vomitoxin(DON)-induced cytotoxicity in L02cells.Methods Human normal hepatocyte L02 were cultured in 1640 medium,and the cells were treated with different concentrations of DON as well as ferroptosis inhibitor Fer-1.qPCR was performed to detect the expression of ferroptosis-related genes induced by DON,C11 BODIPY581/591 fluorescent probe was used to detect the effect of DON on cellular lipid ROS level,and Ferro Orange fluorescent probe was used to detect the effect of DON on intracellular Fe^(2+)level.The effect of Fer-1 on cell viability under DON exposure was detected by CCK-8 assay.Results DON inhibited the expression of ferroptosis-related genes,especially GPX4,SLC7A11 and TFR1,increased cellular lipid ROS and intracellular Fe^(2+)levels,which led to ferroptosis and cell viability reduction.Pretreatment with Fer-1 could inhibit DON-induced cell ferroptosis and increase cell viability.Conclusion DON may induce ferroptosis in L02 cells,and the process is associated with inhibition of SLC7A11/GPX4 pathway.
关 键 词:呕吐毒素 铁死亡 谷胱甘肽过氧化物酶4 溶质载体家族7成员11
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