Fig 1: Exogenous r‐Metrnl treatment attenuated neurological deficits and neuronal degeneration after SAH. (a, b) Rat performance evaluated by modified Garcia scores and beam balance scores at 24 h after SAH. n = 6 per group. (c, d) Representative image and quantitative analyses of FJC at 24 h after SAH. Scale bar = 100 μm, n = 4 per group. Data were represented as mean ± SD. *p < 0.05 vs. Sham; # p < 0.05 vs. SAH + PBS.
Fig 2: Inhibition of C‐KIT or AMPK reversed the anti‐ferroptosis effects of r‐Metrnl after SAH. (a) Representative bands of western blot. (b–i) Quantitative analysis of p‐C‐KIT, C‐KIT, p‐AMPK, AMPK, Nrf2, SLC7A11, GPX4, FSP1, FTH, and ACSL4 expression in the ipsilateral hemisphere at 24 h after SAH. n = 6 per group. (j, k) ELISA analysis of MDA and GSH levels. n = 6 per group. (l, m) Representative image and quantitative analyses of 4HNE at 24 h after SAH. Scale bar = 100 μm, n = 4 per group. Data are represented with mean ± SD. *p < 0.05 vs. Sham, # p < 0.05 vs. SAH + PBS, @ p < 0.05 vs. SAH + r‐Metrnl + DMSO (i.c.v.).
Fig 3: r‐Metrnl administration decreased neuronal degeneration in the hippocampus region 28 days after SAH. (a) Representative microphotograph of Nissl staining showed the location of CA1, CA3, and DG regions within the left hippocampus. (b, c) Representative microphotographs of Nissl staining quantifications of the surviving neurons in CA1, CA3, and DG hippocampal regions. Scale bar = 100 μm, n = 6 per group. Data are represented as mean ± SD. *p < 0.05 vs. Sham group; # p < 0.05 vs. SAH + PBS group.
Fig 4: Inhibition of C‐KIT or AMPK reversed the anti‐ferroptosis effects of r‐Metrnl in Hb‐stimulated primary neurons. (a) Representative bands of western blot. (b–i) Quantitative analysis of p‐C‐KIT, C‐KIT, p‐AMPK, AMPK, Nrf2, SLC7A11, GPX4, FSP1, FTH, and ACSL4 expression in the ipsilateral hemisphere at 24 h after SAH. n = 6 per group. (j, k) ELISA analysis of MDA and GSH levels. n = 6 per group. (l, m) Representative image and quantitative analyses of TMRE. Scale bar = 100 μm, n = 6 per group. Data are represented with mean ± SD. *p < 0.05 vs. Vehicle, # p < 0.05 vs. Hb + PBS, @ p < 0.05 vs. Hb + r‐Metrnl + DMSO.
Fig 5: Endogenous brain Metrnl knockdown exacerbated neuronal ferroptosis after SAH. (a–g) Representative western blot bands and densitometric quantification of Metrnl, SLC7A11, GPX4, FSP1, FTH, and ACSL4. n = 6 per group. (h, i) The MDA and GSH levels. n = 6 per group. (j–m) Representative image and quantitative analyses of GPX4 and 4HNE at 24 h after SAH, scale bar = 100 μm, n = 4 per group. (n, o) Representative images and quantitative analysis of impaired mitochondria observed by transmission electron microscope in each group. Red arrows indicate impaired mitochondria in neurons. Scale bar = 500 nm, n = 4 per group. Data were represented as mean ± SD. *p < 0.05 vs. Sham; # p < 0.05 vs. SAH + scr shRNA.
Supplier Page from OriGene Technologies for Metrnl Rat shRNA Lentiviral Particle (Locus ID 316842)