Figure 5

Combined treatment reduces glial reactivity and protects neurons in the EAE mouse spinal cord. (A) Sections are labeled for GFAP (upper panel), Iba-1 (middle panel), and NeuN (bottom panel) immunoreactivity to detect astrocytes, microglia, and neurons, respectively. Intense GFAP immunoreactivity was present in PBS-treated EAE mice and was reduced in EAE mice treated with hBM-MSCs or minocycline alone, but fewer astroglial cells were activated in the combination-treatment group. Iba-1, which identifies activated microglia, displayed the same immunoreactivity patterns as GFAP in the four EAE groups. The opposite pattern was observed for the neuronal marker, NeuN, in all groups. Scale bar, 200 μm. (B) Stereologic analyses of GFAP fluorescence intensity and (C) the number of Iba-1-positive cells revealed significant reductions in astrocytes and microglial activation in combination-treatment mice compared with those treated with hBM-MSCs or minocycline alone (P < 0.05). (D) Stereologic analysis also revealed a significant increase in the number of NeuN-positive cells in spinal cord sections of EAE mice treated with both hBM-MSCs and minocycline compared with hBM-MSCs or minocycline treatment alone (P < 0.01). Columns, mean; bars, SEM. *P < 0.05, **P < 0.01, ***P < 0.001, one-way ANOVA with post hoc Bonferroni corrections. The results are representative of three independent experiments.