姜黄醇减轻Erastin诱导的小鼠海马神经元铁死亡的研究

Research of Curcumol Alleviating Erastin-induced Ferroptosis in Mouse Hippocampal Neuronal Cells

目的 验证姜黄醇在体外发挥抗铁死亡和细胞保护作用。方法 通过Erastin诱导HT22小鼠海马神经元铁死亡模型,将细胞分为对照组、二甲基亚砜(DMSO)组、Erastin处理组、Erastin+姜黄醇组,检测HT22细胞铁死亡相关蛋白表达水平、细胞内铁离子水平及细胞活力。结果 DMSO组与对照组铁死亡相关蛋白表达水平、铁离子水平、细胞活力基本一致;相比对照组,Erastin组神经元细胞活力显著降低、细胞铁含量显著增加、细胞谷胱甘肽过氧化物酶4(GPX4)活性降低(P<0.05);Erastin+姜黄醇组神经元较Erastin组细胞活力增加、细胞铁含量显著减低、细胞GPX4酶活性增加(P<0.05)。与对照组相比,Erastin组核受体共激活因子4(NCOA4)、长链脂酰辅酶A合成酶4(ASCL4)蛋白水平显著上调,铁蛋白重链1亚基(FTH1)、铁泵蛋白(FPN)、GPX4蛋白水平显著下调(P<0.01);与Erastin组相比,Erastin+姜黄醇组NCOA4、ASCL4蛋白水平显著下调,FTH1、FPN、GPX4蛋白水平显著上调(P<0.01);DMSO组与对照组相比,上述蛋白表达差异无统计学意义(P>0.05)。结论姜黄醇在体外可减轻Erastin诱导的HT22小鼠海马神经元的铁死亡,发挥细胞保护作用。

Objective To verify the anti-ferroptosis and cytoprotective effects of curcumin in vitro.Methods Erastin was used to induce ferroptosis in hippocampal neurons of HT22 mouse.Cells were divided into normal control group,DMSO group,Erastin treatment group and curcumin group.The expression levels of ferroptosis-related proteins in HT22 cells were detected by Western Blot.The intracellular iron level was detected by cell iron ion kit.Cell viability was detected by CCK-8 kit.Results The ferritin-related protein levels,ferrous ion levels and cell viability in DMSO group were basically the same as those in the normal control group.Compared with the normal control group,the cell viability of neurons in Erastin group was significantly decreased(P<0.05),the cell ferrous content was increased(P<0.001),and the cell GPX4 enzyme activity was decreased(P<0.05).Compared with Erastin group,the cell viability of neurons in the curcumin group was increased(P<0.05),the cell ferrous content was decreased(P<0.01),and the GPX4 enzyme activity was increased(P<0.05).Compared with the control group,the protein levels of NCOA4 and ASCL4 in Erastin group were significantly up-regulated,while the protein levels of FTH1,FPN and GPX4 were significantly down-regulated(P<0.01).Compared with the Erastin group,the protein levels of NCOA4 and ASCL4 were significantly down-regulated,while the protein levels of FTH1,FPN and GPX4 were significantly up-regulated in the Erastin and curcumin group(P<0.01).There was no significant difference in the expression of the above proteins between the DMSO group and the control group(P>0.05).Conclusion Curcumin can alleviate erastininduced ferroptosis in hippocampal neurons of HT22 mice in vitro,thus providing cytoprotection.