Fig. 3

Pregabalin inhibited inflammatory response by directly acting on neurons, rather than microglia or astrocyte in vitro. A Q-PCR analysis of Il-6 and Tnf-α mRNA levels in microglia with pregabalin treatment after radiation. B Q-PCR analysis the effects of pregabalin on Il-1β, Tnf-α, and Icam-1 mRNA levels in BV2 cells after radiation. C Schematic diagram of microglia incubated with neuron supernatant. D Q-PCR analysis of Il-1β, Il-6, and Tnf-α mRNA levels in BV2 cells after incubated with the supernatant from pregabalin-treated neurons. E–F Representative immunofluorescent images of IL-6 (E) and TNF-α (F) in BV2 cells among the different groups. Staining with β-tubulin was used to visualize cytoskeleton. G-H The fluorescence intensity data of IL-6 (G) and TNF-α (H) were imaged by confocal microscopy and analyzed. I ELISA analysis of IL-1β protein level in the BV2 cells culture supernatant. Data were analyzed by one-way ANOVA followed by the Student’s t-test analysis. All other groups were compared with the indicated group. n = 3–4 for each group. Data were presented as mean ± SEM, ns = not significant, *p < 0.05, **p < 0.01, and ***p < 0.001