Figure 5.

Sal prevents M1 microglia‐induced neuronal apoptosis (A) Schematic of BV‐2 cells treatments and the coculture system. BV‐2 cells were pre‐treated with Sal for 24 hrs, followed by washing three times and stimulation with LPS for 24 hrs, and then cocultured with PC12 cells. (B, C) TUNEL assay was performed in neurons cocultured with microglia (scale bar: 100 μm). Sal ameliorates neuronal apoptosis induced by M1 microglia. (D) CCK‐8 results pertaining to neurons cocultured with microglia. Cell viability was evidently increased in neurons cocultured with Sal‐pre‐treated microglia. (E, F, G) Fluorescence staining results for MMP, as demonstrated by rhodamine 123 and PI. Representative images of stained neurons from the indicated groups (scale bar: 200 μm). MMP increased significantly, and neuronal apoptosis decreased in cells cocultured with Sal‐pre‐treated microglia. (H, I, J, K) Representative Western blots of and quantitative data for Bcl‐2, Bax, cleaved caspase 3 and β‐actin expression in each group of neurons. The neuronal apoptosis induced by mitochondrial dysfunction was significantly attenuated in cells cocultured with Sal‐pre‐treated microglia. Densitometric analysis of all Western blot bands, whose densities were normalized to those of β‐actin. Data are presented as the mean ± S.D., n = 3 independent experiments. Significant differences between groups are indicated as *P < 0.05 and **P < 0.01.