银杏素通过抑制Nrf2/HO-1/GPX4信号通路诱导神经胶质瘤细胞铁死亡  

Ginkgetin induces ferroptosis of glioma cells by inhibiting Nrf2/HO-1/GPX4 signaling pathway

作　　者：赵印生 鲍龙 孙铁霖 李洋 ZHAO Yinsheng;BAO Long;SUN Tielin;LI Yang(Department of Neurosurgery,the First Affiliated Hospital of Jinzhou Medical University,Liaoning Jinzhou 121000,China)

机构地区：[1]锦州医科大学附属第一医院神经外科,辽宁锦州121000

出　　处：《现代肿瘤医学》2023年第18期3366-3370,共5页Journal of Modern Oncology

基　　金：辽宁省自然科学基金计划项目(编号:2022-BS-313)。

摘　　要：目的:探讨银杏素抑制核因子E2相关因子2(Nrf2)/血红素氧合酶-1(HO-1)/谷胱甘肽过氧化物酶4(GPX4)信号通路,对神经胶质瘤细胞铁死亡的影响。方法:以人神经胶质瘤细胞U251为研究对象,分为对照组、银杏素低(银杏素-L,2.5μmol/L)、中(银杏素-M,5μmol/L)、高(银杏素-H,10μmol/L)浓度组、Nrf2激活剂-SFN(莱菔硫烷,20μmol SFN)组、银杏素-H+SFN(10μmol/L银杏素+20μmol SFN)组;流式细胞仪、CCK-8、DCFH-DA、Western blot分别检测U251细胞的细胞凋亡率、增殖水平、活性氧(ROS)含量以及Nrf2/HO-1/GPX4通路蛋白表达水平;铁离子检测试剂检测细胞内铁含量;透射电镜观察细胞超微结构。结果:与对照组相比,银杏素-L、银杏素-M、银杏素-H组线粒体膜厚度及嵴改变,增殖率、Nrf2、HO-1、GPX4蛋白表达均显著下降,凋亡率、ROS及铁含量显著增加(P<0.05);与银杏素-H组相比,银杏素-H+SFN组增殖率、Nrf2、HO-1、GPX4蛋白表达均显著增加,凋亡率、ROS及铁含量显著下降(P<0.05);与SFN组相比,银杏素-H+SFN组增殖率、Nrf2、HO-1、GPX4蛋白表达均显著下降,凋亡率、ROS及铁含量显著增加(P<0.05)。结论:银杏素通过抑制Nrf2/HO-1/GPX4信号通路,可以诱导神经胶质瘤细胞铁死亡,进而发挥抗肿瘤的作用。Objective:To investigate the impact of Ginkgetin on ferroptosis of glioma cells by inhibiting nuclear factor E2-related factor 2(Nrf2)/heme oxygenase-1(HO-1)/glutathione peroxidase 4(GPX4)signaling pathway.Methods:Human glioma cells U251 were regarded as the research objects,and they were separated into control group,low(Ginkgetin-L,2.5μmol/L),medium(Ginkgetin-M,5μmol/L),and high(Ginkgetin-H,10μmol/L)concentration Ginkgetin group,Nrf2 activator-SFN(sulforaphane,20μmol SFN)group,and Ginkgetin-H+SFN(10μmol/L Ginkgetin+20μmol SFN)group.Flow cytometry,CCK-8,DCFH-DA and Western blot were performed to detect the apoptosis rate,proliferation level,reactive oxygen species(ROS)content and Nrf2/HO-1/GPX4 pathway protein expression level of U251 cells,respectively.Iron ion detection reagent was applied to detect intracellular iron content.Transmission electron microscope was used to observe cell ultrastructure.Results:Compared with the control group,the mitochondrial membrane thickness and ridge changed in Ginkgetin-L,Ginkgetin-M,and Ginkgetin-H groups,the proliferation rate,Nrf2,HO-1,and GPX4 protein expressions decreased obviously,the apoptosis rate,contents of ROS and iron increased obviously(P<0.05).Compared with the Ginkgetin-H group,the proliferation rate,Nrf2,HO-1,and GPX4 protein expressions increased obviously in Ginkgetin-H+SFN group,the apoptosis rate,contents of ROS and iron decreased obviously(P<0.05).Compared with the SFN group,the proliferation rate,Nrf2,HO-1,and GPX4 protein expressions decreased obviously in Ginkgetin-H+SFN group,the apoptosis rate,contents of ROS and iron increased obviously(P<0.05).Conclusion:Ginkgetin can induce ferroptosis in glioma cells by inhibiting the Nrf2/HO-1/GPX4 signaling pathway,thereby exerting an anti-tumor effect.

关 键 词：核因子E2相关因子2/血红素氧合酶-1/谷胱甘肽过氧化物酶4 银杏素 神经胶质瘤 铁死亡 

分 类 号：R730.264[医药卫生—肿瘤]