MPV17 inhibits iron overload-induced ferroptosis of splenic CD3+T cells in mice by blocking ERK pathway
Objective This work aimed to explore the effect of iron overload on splenic injury and the role of MPV17 in the ferroptosis of splenic CD3+T cells from mice subjected to iron overload.Methods Mice were randomly divided into normal diet group,high-iron diet group,high-iron diet combined with Fer-1 treatment group,and high-iron diet combined with adenovirus harboring MPV17 injection group,with 5 mice in each group.After treatment for 8 weeks,mice spleens were harvested and fixed;Histological section and HE staining were performed to observe the structures of the spleens;Cell death of CD3+T cells was detected by propidium iodide(PI)staining;The lipid peroxidation levels were detected by C11 BODIPY581/591 staining;The mRNA levels of Solute carrier family 7 member 11(SLC7A11)and prostaglandin-endoperoxide synthase 2(PTGS2)were detected by qPCR assays;The macrophage phenotype-switching(M1/M2)were detected by flow cytometry;The levels of TNF-α,IL-1β and IL-6 were measured by ELISA assays.Moreover,high-iron diet combined with extracellular signal-regulated kinase(ERK)inhibitor treatment group,ERK agonist treatment group,β-gal combined with ERK agonist treatment group,and MPV17 overexpression combined with ERK agonist treatment group were added.The protein levels of MPV17,glutathione peroxidase 4(GPX4)and phosphorylated ERK(p-ERK)were detected by Western blot;The mitochondrial membrane potential was detected by JC-1 staining and flow cytometry.Results Compared with the normal diet group,the red pulps of the mice spleens from the high-iron diet group showed irregular structures and the white pulps were almost missing;Cell death,lipid peroxides,and the expression levels of SLC7A11 and PTGS2 increased;Both the ratio of M1 macrophages to M2 macrophages and the levels of inflammatory factors increased.Fer-1 treatment or overexpression of MPV17 in the high-iron diet mice group partially recovered the irregular structures of the spleens,reduced cell death and lipid peroxides in CD3+T cells,and decreased the expression levels of SLC7A11 and PTGS2;The ratio of M1/M2 macrophages and the levels of inflammatory factors were decreased.High-iron diet decreased the protein levels of GPX4 while p-ERK were up-regulated.Inhibition of ERK partially recovered the protein levels of GPX4;ERK agonist decreased the protein levels of GPX4;MPV17 inhibited the ERK signaling and partially recovered the protein levels of GPX4 and the decreased mitochondrial membrane potential of CD3+T induced by ERK activation.Conclusion Iron overload resulted in splenic injury and ferroptosis in the splenic CD3+T cells;MPV17 prevented splenic injury and ferroptosis of splenic CD3'T cells of the iron overload mice through blocking ERK signaling pathway.
iron overloadCD3+T cellferroptosismitochondrial inner membrane protein 17(MPV17)extracellular signal-regulated kinases(ERK)
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