Figure 8

SIRT1 mediates the neuroprotective effect of NeuroHeal. (A) Diagram of the mechanisms of action of spermidine and Ex-527 and bar graph of the percentage of avulsed MNs with high nuclear immunofluorescence intensity for each marker on the ipsilateral side of RA animals treated with different drugs (n = 6 for untreated; n = 3 for Veh DMSO; n = 4 other groups, ANOVA, post hoc Bonferroni*p < 0.05 vs, ^#p < 0.05 vs. C1, ^$p < 0.05 vs. C1 + Sperm, ^&p < 0.05 vs. Sperm). (B) Representative microphotographs of MNs on the ipsilateral sides and associated histogram of the average percentage of MN survival ± SEM in animals intrathecally treated with spermidine (Sperm) or Ex-527 with or without NeuroHeal (C1) (n = 4, ANOVA, post hoc Bonferroni *p < 0.05 vs. untreated, ^#p < 0.05 vs. veh, ^$p < 0.05 vs. C1 + Ex-527, *p < 0.05). Scale bar = 100 μm. (C) Microphotographs of GAP43 immunostaining at the ventral horns of the ipsilateral sides from animals treated with either AAVrh10-GFP, AAVrh10-SIRT1 or C1. Scale bar = 100 µm. Bar graph of the average immunoreactivity in a fixed region of interest of the white matter (n = 3–4, *p < 0.05 vs. AAVrh10-GFP). (D) Representative microphotographs of astrocyte (GFAP) or microglia (Iba1) staining at the ventral horns of the ipsilateral sides of RA animals treated with either NeuroHeal (C1) or spermidine (Sperm). Scale bar = 100 μm. Associated bar graphs of the average immunoreactivity in a fixed region of interest of the grey matter (*p < 0.05 vs. C1).