Fig. 4

The effects on neuronal survival following inhibition of the synthesis of glutamate, reactive oxygen species, and inflammatory factors. Following 6 h of re-oxygenation, we added 6-diazo-5-oxo-l-norleucine (DON), anti-TNF-α antibody (aTNF), anti-IL-1β receptor antibody (IL-1ra), and apocynin to neuron-astrocyte co-cultures that were treated with oxygen-glucose deprivation (OGD) to ensure blockade of the synthesis of glutamate, TNF-α, IL-1β, and reactive oxygen species (ROS), respectively. We assessed alterations in cell numbers in the four groups at 1, 3, and 6 h after OGD. a Representative deconvolution fluorescence images of neuron-astrocyte co-cultures. Neurons were stained with an anti-microtubule associated protein II (MAP-II) antibody (green). Astrocytes were stained with an anti-glial fibrillary acidic protein (GFAP) antibody (red). All scale bars represent 100 μm. b The relative neuronal survival rate was quantified. The viability of non-treated neurons was normalized to 1.0. Survival rate of the remaining groups divided by the survival rate of NT group to calculate the relative survival rate. To assess the effect of IP and various drugs on neuronal survival after OGD/R, we calculated the statistical differences between each drug intervention group and the OGD/R group, respectively. Each column indicates the mean ± SEM (n = 5). ***P < 0.001 when compared to the survival rate of OGD/R co-cultures