Fig. 7

The combination of MS-275 and resveratrol reduces microglia toxicity and activation promoted by NCM. (a) LDH release from mixed glial cultures exposed to NCM. Mixed glial cultures were incubated overnight with NCM obtained from neurons subjected to OGD (NCM-OGD) or from control neurons (NCM-control). NCM-OGD induced significant toxicity in glial cells. The association of MS-275 (0.1 μM) and resveratrol (3 μM) significantly reduced the cellular damage mediated by NCM-OGD. Each value is expressed as the mean ± s.e.m. of 5 different replicate experiments, each performed in triplicate. *p<0.05, one-way ANOVA followed by Tukey multiple comparison test. (b) LDH release from pure astrocyte cultures exposed to NCM. Primary astrocytes were incubated overnight with NCM-OGD or NCM-control. The exposure to NCM did not increase the release of LDH from astrocytes. Each value is expressed as the mean ± s.e.m. of 3 different replicate experiments, each performed in triplicate. p>0.05, one-way ANOVA followed by Tukey multiple comparison test. (c, d) Representative images of (c) Iba1 (green) and GFAP (red) and (d) Iba1 (green) and iNOS (red) immunofluorescence staining on mixed glial cells incubated with NCM-OGD or NCM-control. Cultures incubated overnight with NCM-OGD displayed increased immunofluorescence for iNOS and morphological signs of activation in GFAP+ astrocytes and Iba1+ microglial cells. In (d), arrows in NCM-OGD image represent sites of co-reactivity to iNOS and Iba1. Treatment with MS-275 (0.1 μM) and resveratrol (3 μM) reduced reactivity to iNOS, and promoted recovery of the signs of glial cells activation in Iba1+ and GFAP+ cells. Images are representative of 3 experiments per group. Scale bar: in c = 50 μm for c and d.