The effects of i.t. Ad-PTEN on CCI-induced microglial and astrocytic activation and upregulation of TNF-α. Spinal cord sections (10 μm) obtained 14 days post-surgery from sham-operated plus i.t. vehicle (A, D, G), CCI plus i.t. Ad-GFP (B, E, H), and CCI plus i.t. Ad-PTEN (C, F, I) groups. Immunostaining images show cells labeled with OX-42 (red; (A-C)) and GFAP (red; (D-F)), TNF-α (red; (G-I)) in the spinal cord. Quantification of OX-42 (J) and GFAP (K), and TNF-α (L) immunoreactivity in the ipsilateral dorsal horn of the lumbar spinal gray matter. Ad-GFP, adenovirus-mediated green fluorescent protein; Ad-PTEN, adenovirus-mediated phosphatase and tensin homolog deleted from chromosome 10; CCI, chronic constriction injury; GFAP, glial fibrillary acidic protein; TNF, tumor necrosis factor. Each bar in (J-L) represents the mean ± SEM with six rats per group. Ad-PTEN (i.t.) significantly inhibited CCI-induced upregulation of spinal OX-42, GFAP, and TNF-α immunoreactivity. Scale bars: 200 μm for all images (A-I). *P < 0.05 compared with sham-operated plus i.t. vehicle group; #
P < 0.05 compared with CCI plus i.t. Ad-GFP.