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Fig. 5 | Journal of Neuroinflammation

Fig. 5

From: Dimethyl fumarate attenuates experimental autoimmune neuritis through the nuclear factor erythroid-derived 2-related factor 2/hemoxygenase-1 pathway by altering the balance of M1/M2 macrophages

Fig. 5

Preventative and therapeutic DMF treatment increases the production of Nrf2, the percentage of Nrf2 in macrophages, and the level of HO-1 in sciatic nerves of EAN rats. DMF was administered orally to EAN rats (n = 6 in each group) from day 1 to day 16 and day 7 to day 16 post-immunization for preventative and therapeutic treatment, respectively. On day 16 post-immunization, sciatic nerves of rats from each group were harvested for fluorescence immunohistochemistry and Western blotting. DAPI counterstaining was used in tissue sections for cellular identification. a Fluorescence photomicrographs of anti-Nrf2 (green), anti-CD68 (red), and anti-HO-1 (red) staining. In both the DMF-preventative and DMF-therapeutic groups, Nrf2 and HO-1 stainings were more intense and robust when compared to the CMC group. Co-staining macrophages with Nrf2 showed that DMF-treated groups had a higher radio of Nrf2 positive macrophages than CMC group. Scale bar is 20 μm. b Quantitation of cellular anti-Nrf2 staining. There were significantly more Nrf2+ cells per square millimeter in the DMF-treated groups compared to the CMC group. c Fluorescence photomicrographs and quantitation of anti-Nrf2 and anti-CD68 staining showed that in both the DMF-preventative and DMF-therapeutic groups, the percentage of Nrf2- and CD68-positive cells in CD68-positive cells was higher when compared to the CMC group. d Quantitation of cellular anti-HO-1 staining. There was significantly more HO-1+ cells in the DMF-treated groups compared to the CMC group. e Western blotting of Nrf2 and HO-1 protein compared to β-actin standard. DMF-preventative and DMF-therapeutic groups both showed increased expression of Nrf2 and HO-1 in sciatic nerves when compared to the CMC group. f, g Quantitation of Western blotting of Nrf2 and HO-1. Relative expression of Nrf2 and HO-1 in DMF-treated groups was significantly greater compared to the CMC group. Comparisons between the CMC group and DMF-treated groups were made by Mann–Whitney U test. Results are expressed as means ± SEM (*p < 0.05 compared to the CMC control group; n = 4–5). The experiment was repeated 3 times with similar results

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