SB225002 treatment inhibits L5-SNL-induced microglia proliferation and activation and the pro-inflammatory response. (A) The number of microglia were analyzed by immunofluorescence in the spinal cord of L5-SNL rats with or without SB225002 treatment compared with sham-operated group. All samples were tested 3 times, and data were presented as mean ± SD. (B) mRNA expression of Iba-1 was assessed by qPCR analysis in spinal microglia isolated from L5-SNL rats with or without SB225002 treatment compared with sham-operated group. (C and D) Western blot (C) and quantitative analysis (D) of Iba-1 protein expression in spinal microglia isolated from L5-SNL rats with or without SB225002 treatment compared with sham-operated group (n=3 of each group). GAPDH was used as loading control in western blot analysis. (E-G) The levels of pro-inflammatory cytokines TNF-α (E), IL-1β (F) and IL-6 (G) were determined by ELISA assay using ELISA kits in spinal cord tissues of L5-SNL rats with or without SB225002 treatment compared with sham-operated group. *P<0.05, **P<0.01. L5-SNL, L5 spinal nerve ligation; TNF, tumor necrosis factor; IL, interleukin; Iba-1, ionized calcium binding adaptor molecule-1.