去铁胺在麦芽酚铝诱导大鼠肾上腺嗜铬细胞瘤细胞铁死亡中的作用

The role of DFO in Al(mal)3-induced ferroptosis in PC12 cells

的探讨麦芽酚铝[Al(mal)3]诱导大鼠肾上腺嗜铬细胞瘤(PC12)细胞铁死亡及去铁胺(DFO)在铁死亡发生过程中的作用机制。方法将PC12细胞分为对照组、Al(mal)3组、DMSO组、DFO组、Al(mal)3+DMSO组和Al(mal)3+DFO组,在DMSO组和Al(mal)3+DMSO组、DFO组和Al(mal)3+DFO组中分别加入DMSO和DFO预孵育2 h,后在Al(mal)3组、Al(mal)3+DMSO组和Al(mal)3+DFO组中加入Al(mal)3至终浓度为200μmol/L。采用CCK8法检测细胞存活率,倒置显微镜观察细胞形态和活性氧(ROS)水平,比色法检测细胞增殖毒性和细胞内铁离子含量,生化法检测细胞还原型谷胱甘肽(GSH)含量和谷胱甘肽过氧化物酶(GSH-PX)活力。结果与对照组比较,Al(mal)3组PC12细胞存活率明显降低,乳酸脱氢酶(LDH)活力和铁离子含量明显升高,GSH含量和GSH-PX活力明显降低,差异均有统计学意义(P<0.05)。与Al(mal)3+DMSO组比较,Al(mal)3+DFO组PC12细胞存活率明显提高,LDH活力明显降低,GSH含量和GSH-PX活力明显升高,差异均有统计学意义(P<0.05)。Al(mal)3+DFO组细胞内铁离子含量有所降低,但差异无统计学意义(P>0.05)。与DMSO组比较,DFO组细胞内铁离子含量较低,差异有统计学意义(P<0.05)。与对照组比较,Al(mal)3染毒组ROS荧光增强,荧光细胞数目多;加入DFO后,与Al(mal)3+DMSO组比较,细胞内ROS荧光减弱,荧光细胞数目减少。结论Al(mal)3可导致PC12细胞发生铁死亡,DFO可通过降低细胞内铁离子含量和增加细胞抗氧化损伤能力而抑制铁死亡发生。

Objective To investigate the mechanism of Al(mal)3-induced ferroptosis in rat adrenal pheochromocytoma cells(PC12),to explore the effect of deferoxamine(DFO).Methods Taken PC12 cells growing at logarithmic phase and divided into 6 groups:control group,200μmol/L Al(mal)3 group,0.5%DMSO group,200μmol/L DFO group,Al(mal)3+DMSO group,Al(mal)3+DFO group.DMSO and DFO were added to the DMSO group and the Al(mal)3+DMSO group,the DFO group and the Al(mal)3+DFO group for 2 h,respectively,Al(mal)3 was then added to the Al(mal)3 group,Al(mal)3+DMSO group,and the Al(mal)3+DFO group to a final concentration of 200μmol/L.The cell viability was detected by CCK8,the morphology and ROS levels of PC12 cells was observed by inverted microscope,the cell proliferation toxicity and intracellular iron ion content were detected by colorimetry,the GSH content and GSH-PX activity were detected by biochemical method.Results Al(mal)3 exposure significantly inhibited the growth of PC12 cells and destroyed the cell morphological structure,resulting in increased LDH activity and intracellular iron ion content in PC12 cells,decreased GSH content and GSH-PX activity,increased ROS levels;the combined treatment of Al(mal)3+DFO can significantly improve the cell viability of PC12 cells,improved cell morphology,decreased cell LDH activity and intracellular iron ion content(P>0.05),increased GSH content and GSH-PX activity,decreased ROS levels.Conclusion Al(mal)3 can induce ferroptosis in PC12 cells,DFO may inhibit ferroptosis by reducing intracellular iron levels and reducing oxidative damage.