Hepcidin Promoted Ferroptosis through Iron Metabolism which Is Associated with DMT1 Signaling Activation in Early Brain Injury following Subarachnoid Hemorrhage

Figure 3

Immunofluorescence staining for hepcidin, DMT1 and GPX4 showed co-localization in neurons 24 h after SAH. Hepcidin, DMT1 and GPX4 were co-localized with NeuN-positive cells (neurons); hepcidin (red), DMT1 (red), GPX4 (red) NeuN (green), DAPI (nucleus, blue); original magnification 400× (a) (). Expression of hepcidin, DMT1, FPN1 and GPX4 at different time points after SAH. Representative WBs of hepcidin, DMT1, FPN1 and GPX4 (b). Densitometric quantification of the optical densities of these protein bands (c-f). All protein expression levels were significantly changed at 24-72 h after SAH, P <0.05 vs the sham group (, each group). Expression of DMT1 and GPX4 after different doses of ebselen were introduced following SAH. Representative WBs of DMT1 and GPX4 (G), densitometric quantification of the optical densities of these protein bands (h-i), all protein expression levels were significantly changed at the dose of 4 mg/kg ebselen after SAH, P <0.05 vs the Abs+SAH group, (, each group). Perl’s staining of iron after 4 mg/kg ebselen was administered. Positive areas 4 mg/kg ebselen decreased, compared with the Abs+SAH groups, final magnification 400× (j) (, each group). The activity of components of ferroptosis (iron content, MDA, GSH, and GPX4) were determined after 4 mg/kg ebselen was introduced. Treatment with 4 mg/kg ebselen decreased MDA while increasing GSH and GPX4 activity and protecting against ferroptosis, compared with the Abs+SAH groups (k-n), P <0.05 vs the Abs+SAH group (, each group).

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