江苏大学学报（医学版）2025，Vol.35Issue(4)：304-310,317,8.DOI:10.13312/j.issn.1671-7783.y240027
神经干细胞分泌组对人神经母细胞瘤SH-SY5Y细胞铁死亡的抑制作用及其机制
The inhibitory effect and mechanism of neural stem cell secretome on ferroptosis in human neuroblastoma SH-SY5Y cells
摘要
Abstract
Objective:To explore the effect of neural stem cell secretome(NSC-S)on ferroptosis human neuroblastoma SH-SY5Y cells and its underlying mechanism.Methods:Human neuroblastoma SH-SY5Y cells were used to establish neuronal ferroptosis model in vitro.And the cells were divided into three groups:control group,Erastin group and Erastin+NSC-S group;cells were treated with DMEM,DMEM containing 10 μmol/L Erastin,and NSC-S containing 10 μmol/L Erastin,respectively.The activities of lactate dehydrogenase(LDH),intracellular reactive oxygen species and malondialdehyde in the supernatants of cells in each group were detected by commercial kits,and the contents of free ferrous ions were detected by fluorescent probes.The expression level of glutathione peroxidase 4(GPX4)protein in cells was determined by Western blotting.The components of NSC-S were detected by liquid chromatography tandem mass spectrometry(LC-MS/MS)and bioinformatics analysis and screening were conducted.In addition,SH-SY5Y cells were divided into four groups,which were control group,Erastin group,Erastin+NSC-S group,and Erastin+Parkinson's disease protein 7(PARK7)group.And cells were treated with DMEM,DMEM containing 10 μmol/L Erastin,NSC-S containing 10 μmol/L Erastin,and DMEM containing 200 μg/L PARK7+10 μmol/L Erastin,respectively.The viability of SH-SY5Y cells was detected by the CCK-8 assay,and the contents of reactive oxygen species,malondialdehyde,ferrous ions in the cells as well as the expression level of GPX4 protein were also detected.Results:Compared with the Erastin group,the LDH activity of SH-SY5Y cells supernatant in the Erastin+NSC-S group was significantly decreased(P<0.001),the contents of intracellular reactive oxygen species,malondialdehyde and ferrous ions were significantly reduced(P<0.001 or P<0.05),and the relative expression level of GPX4 protein was greatly increased(P<0.05).LC-MS/MS analysis revealed that NSC-S contained PARK7 protein.Compared with the Erastin group,the viability of SH-SY5Y cells in the Erastin+PARK7 group was significantly enhanced(P<0.001),the contents of reactive oxygen species,malondialdehyde and ferrous ions were significantly decreased(P<0.001),and the relative expression level of GPX4 protein was significantly increased(P<0.05).Conclusion:NSC-S could inhibit ferroptosis induced by Erastin in human neuroblastoma SH-SY5Y cells,which may be related to the mediation of PARK7 in NSC-S.关键词
神经干细胞/人神经母细胞瘤/神经元/铁死亡/帕金森病蛋白7Key words
neural stem cells/human neuroblastoma/neuron/ferroptosis/PARK7分类
医药卫生引用本文复制引用
曾宇,仇昀,司雨,郭涛,倪银莹,张婧雯,王子瑜,陈波,胡嘉波..神经干细胞分泌组对人神经母细胞瘤SH-SY5Y细胞铁死亡的抑制作用及其机制[J].江苏大学学报（医学版）,2025,35(4):304-310,317,8.基金项目
校企协同创新项目(20200605) （20200605）
