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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