Ferrostatin-1抑制铁死亡缓解阿霉素诱导的足细胞损伤

Ferrostatin-1 alleviates adriamycin-induced podocyte injury by inhibits ferroptosis

目的探讨ferrostatin-1(Fer-1)在阿霉素(adriamycin,ADR)诱导的肾小球足细胞铁死亡的影响。方法体外ADR刺激足细胞,Western blot及RT-qPCR检测ADR组及Fer-1+ADR组足细胞的铁死亡信号通路及足细胞损伤相关蛋白的表达;CCK-8检测足细胞活性;丙二醛(malondialdehyde,MDA)、谷胱甘肽(glutathione,GSH)、铁池(Fe^(2+))试剂盒检测相关铁死亡指标。体内构建ADR肾病小鼠模型,Western blot及RT-qPCR检测小鼠肾小球足细胞的铁死亡信号通路相关蛋白的表达情况,通过4-HNE免疫组化评估肾脏铁死亡水平。结果ADR刺激下足细胞铁死亡相关标志物ACSL4蛋白表达量明显上调,GPX4和SLC7A11的蛋白表达水平明显下调。与ADR组相比,Fer-1干预下可以部分恢复足细胞铁死亡基因的异常表达,同时改善足细胞损伤蛋白Desmin表达水平。MDA、GSH、Fe^(2+)检测结果显示Fer-1改善足细胞脂质过氧化水平。ADR模型组小鼠ACSL4蛋白表达量明显上调,GPX4和SLC7A11的蛋白表达水平明显上调,铁死亡标志物4-HNE染色阳性增加。结论阿霉素诱导铁死亡加重足细胞损伤,抑制铁死亡可能是防治阿霉素肾脏病进展的新策略。

Aim To investigate the role of ferrostatin-1(Fer-1)in adriamycin(ADR)induced podocyte injury.Methods Western blot and RT-qPCR were used to detect the expression of the ferroptosis signaling pathway and podocyte injury-related proteins in podocytes treated with ADR and Fer-1.The therapeutic effect of Fer-1 on podocytes was screened by CCK-8;malondialdehyde(MDA),glutathione(GSH)and Fe^(2+)kit were used to evaluate the lipid peroxidation level of podocytes.A mouse model of ADR nephropathy was constructed in vivo,and the expression of ferroptosis related proteins in mouse glomerular podocytes was detected by Western blot and RT-qPCR.The ferroptosis level of kidney was evaluated by immunohistochemistry staining of 4-HNE.Results The expression level of ACSL4,a marker related to ferroptosis,was significantly up-regulated.GPX4 and SLC7A11 were significantly down-regulated in podocytes under ADR stimulation.Compared with the ADR group,Fer-1 intervention could partially restore the anomalous change of ferroptosis related protein and improve the expression level of podocellular damage protein desmin.The results of MDA,GSH and Fe^(2+)showed that Fer-1 could improve the lipid peroxidation level of podocytes.The expression level of ACSL4 was significantly up-regulated and GPX4 and SLC7A11 were significantly up-regulated in the ADR model group.IHC staining of 4-HNE increased in ADR model group.Conclusions Adriamycin promotes podocyte injury by induced ferroptosis,and inhibiting ferroptosis may be a potential strategy to prevent the progression of ADR nephropathy.