去铁胺在甲基汞诱导细胞铁死亡中的作用及其机制  

The role of deferoxamine in methylmercury-induced ferroptosis and its mechanism

作　　者：刘婷婷 董丽华 张宇[1] 徐伟 朱海涛[3] 李芳 王苏华[1] 邢光伟[1] 陆荣柱[1] LIU Ting-ting;DONG Li-hua;ZHANG Yu;XU Wei;ZHU Hai-tao;LI Fang;WANG Su-hua;XING Guang-wei;LU Rong-zhu(School of Medicine, Jiangsu University, Zhenjiang Jiangsu 212013;Department of Laboratory, Changzhou First People′s Hospital, Changzhou Jiangsu 213000;Department of Imaging, Affiliated Hospital of Jiangsu University, Zhenjiang Jiangsu 212001, China)

机构地区：[1]江苏大学医学院,江苏镇江212013 [2]常州市第一人民医院检验科,江苏常州213000 [3]江苏大学附属医院医学影像科,江苏镇江212001

出　　处：《江苏大学学报（医学版）》2020年第6期504-509,共6页Journal of Jiangsu University：Medicine Edition

基　　金：国家自然科学基金资助项目(30872139,81273124,31100964)。

摘　　要：目的:探讨去铁胺预处理对甲基汞毒性作用的影响及其潜在的机制。方法:选用大鼠肾上腺嗜铬细胞瘤PC12细胞与大鼠肝BRL细胞,对照组用含10%胎牛血清的高糖培养基培养0.5 h,甲基汞组用不同浓度甲基汞(1.0、2.5、5.0、10.0μmol/L)处理0.5 h,MTT法检测细胞活力,免疫印迹法检测谷胱甘肽过氧化物酶4(glutathione peroxidase 4,GPx4)蛋白表达,筛选最适浓度甲基汞。另取PC12细胞与BRL细胞,对照组用含10%胎牛血清的高糖培养基培养6.5 h,去铁胺+甲基汞组分别用0、50、100、200、400μmol/L去铁胺预处理6 h,再用10.0μmol/L甲基汞处理0.5 h,免疫印迹法检测GPx4及缺氧诱导因子-1α(hypoxia inducible factor-1α,HIF-1α)蛋白表达,MTT法检测细胞活力。结果:与对照组相比,10.0μmol/L甲基汞组两种细胞活力降低最显著,以此浓度进行后续实验;随着甲基汞浓度逐渐增加,两种细胞中GPx4表达逐渐降低。与0μmol/L去铁胺+甲基汞组比较,随着去铁胺浓度增加,两种细胞中GPx4和HIF-1α蛋白表达逐渐增加;400μmol/L去铁胺+甲基汞组PC12细胞活力明显增加,200μmol/L去铁胺+甲基汞组BRL细胞活力明显增加。结论:去铁胺可拮抗甲基汞致细胞的铁死亡,可能与上调HIF-1α及铁死亡关键调控因子GPx4表达相关。Objective:To investigate the role of deferoxamine pretreatment on the toxicity of methylmercury and its potential mechanism.Methods:PC12 and BRL cells were selected and divided into groups:control group,cells were cultured with high sugar medium containing with 10%fetal bovine serum for 0.5 h;methylmercury group,cells were treated with different concentrations(1.0,2.5,5.0,10.0μmol/L)of methylmercury for 0.5 h,MTT assay was used to detect cell viability,Western blotting was used to detect the expression of glutathione peroxidase 4(GPx4).The above two results were used to screen the optimal concentration of methylmercury.In addition,PC12 cells and BRL cells were divided into control group which cells were cultured with high sugar medium containing with 10%fetal bovine serum for 6.5 h,and different concentrations of deferoxamine+methylmercury groups which cells were pretreated with 0,50,100,200 and 400μmol/L deferoxamine for 6 h,then treated with methylmercury(10.0μmol/L)for 0.5 h,Western blotting assay was used to detect the expression of GPx4 and hypoxia inducible factor-1α(HIF-1α),MTT assay was used to detect cell viability.Results:Compared with the control group,10.0μmol/L methylmercury group was shown with significantly decreased cell viability in both cell lines,therefore,10.0μmol/L methylmercury was used for subsequent experiments.Compared with 0μmol/L deferamine+methylmercury group,the expression of GPx4 and HIF-1αenhanced with the increase of deferamine concentration in both cell lines;cell viability was enhanced when deferoxamine concentration reached to 400μmol/L in PC12 cells and 200μmol/L in BRL cells.Conclusion:Deferoxamine may antagonize methylmercury-induced ferroptosis through upregulating the expression of HIF-1αand GPx4.

关 键 词：甲基汞 铁死亡 去铁胺 缺氧诱导因子-1Α 谷胱甘肽过氧化物酶4 

分 类 号：R393[医药卫生—基础医学]