原花青素通过Nrf2抑制Erastin介导的细胞铁死亡

PROANTHOCYANIDINS INHIBIT ERASTIN-INDUCED CELL FERROPTOSIS VIA NRF2

目的探讨原花青素(proanthocyanidins,PC)对Erastin诱导的人神经母细胞瘤SH-SY5Y细胞铁死亡的影响及作用机制。方法通过MTT法检测细胞活力。采用试剂盒测定细胞内铁离子水平、活性氧(reactive oxygen species,ROS)水平、丙二醛(malondialdehyde,MDA)含量、谷胱甘肽(glutathione,GSH)水平和超氧化物歧化酶(superoxide dismutase,SOD)活性。通过蛋白质免疫印迹法检测细胞内溶质载体家族7成员11(solute carrier family 7 member 11,SLC7A11)、谷胱甘肽过氧化物酶4(glutathione peroxidase 4,GPX4)、核转录因子E2相关因子2(nuclear factor erythroid2-related factor 2,Nfr2)和血红素氧合酶1(heme oxygenase-1,HO-1)的蛋白表达水平。结果PC预处理能够保护SH-SY5Y细胞免受Erastin诱导的细胞铁死亡,主要是通过抑制ROS生成、铁离子水平和MDA含量升高,上调GSH水平和SOD活性保护细胞免受氧化损伤,增强细胞的抗氧化能力。此外,PC还可以通过激活Nrf2/HO-1信号通路上调SLC7A11、GPX4的表达,而Nrf2抑制剂ML385显著消除了PC对细胞内铁死亡的抑制作用,SLC7A11、GPX4、Nrf2和HO-1的蛋白表达降低。结论PC可以抑制Erastin诱导的SH-SY5Y细胞铁死亡,从而发挥其神经保护作用,其潜在机制可能与Nrf2/HO-1信号通路有关。

Objective To investigate the effect of proanthocyanidins(PC)on erastin-induced ferroptosis in human neuroblastoma SH-SY5Y cells and the mechanism of action.Methods Cell viability was detected by thiazolyl blue tetrazolium bromide(MTT)colorimetric method.Intracellular iron ion level,reactive oxygen species(ROS)level,malondialdehyde(MDA)content,glutathione(GSH)level and superoxide dismutase(SOD)activity were detected by corresponding kits.The protein expression levels of intracellular solute carrier family 7 member 11(SLC7A11),glutathione peroxidase 4(GPX4),nuclear factor erythroid 2-related factor 2(Nfr2)and heme oxygenase 1(HO-1)were measured by Western blot assay.Results PC pretreatment was able to protect SH-SY5Y cells from erastin-induced cellular ferroptosis,mainly by inhibiting ROS production,elevated iron ion levels and MDA production,upregulating GSH levels and SOD activity.In addition,PC could also upregulate the expressions of SLC7A11 and GPX4 by activating the Nrf2/HO-1 signaling pathway,while the Nrf2 inhibitor ML385 significantly abolished the inhibitory effect of PC on intracellular ferroptosis,and the protein expressions of SLC7A11,GPX4,Nrf2 and HO-1 were decreased.Conclusion PC can inhibit erastin-induced ferroptosis,thus exerting its neuroprotective effect,and the underlying mechanism may be related to the Nrf2/HO-1signaling pathway.