19; P = 0.0067 and ANOVA, F(1,8) = 7.903; P = 0.0228, respectively). IL-1β To replicate our results described in Figure 3D and 3F, separate L4–L6 lumbar spinal cord tissue sections were

processed and analyzed. Compared to non-neuropathic sham-operated rats given i.t. AM1241 or equivolume vehicle, CCI-induced neuropathy produced a robust unilateral increase in dorsal horn IL-1β IR (ANOVA, F(1,8) = 10.46; P = 0.0120), while, compared to controls, Inhibitors,research,lifescience,medical no differences in contralateral IL-1β were observed (ANOVA, F(1,8) = 1.627; P = 0.2379) (Fig. 4E and 4F). Conversely, following AM1241 administration, significantly lower levels of IL-1β IR were detected (ANOVA, F(1,8) = 9.431; P = 0.0153). IL-1β IR observed in the contralateral dorsal horn was not substantially elevated when compared to vehicle-injected Inhibitors,research,lifescience,medical animals (ANOVA, F(1,8) = 1.321; P = 0.2836). p38-MAPK and DAPI Spinal p-p38MAPK is widely characterized to click here mediate allodynia through the actions of spinal IL-1β (Ji and Suter 2007; Ji et al. 2009). Therefore, p-p38MAPK Inhibitors,research,lifescience,medical was examined. Compared to non-neuropathic sham-operated rats given i.t. AM1241 or equivolume

vehicle, CCI-induced neuropathy produced a robust p-p38MAPK bilateral IR increase in the spinal cord dorsal horn (ANOVA, F(1,8) = 223.1; P < 0.0001 and ANOVA, F(1,8) = 148.0; P < 0.0001, respectively) (Fig. 4G and 4H). In contrast, tissues from rats treated with i.t. AM1241 revealed dramatically lower levels of p-p38MAPK IR Inhibitors,research,lifescience,medical that were close to or similar to spinal cord tissues from non-neuropathic sham-treated rats (ANOVA, F(1,8) = 85.82; P < 0.0001 and ANOVA, F(1,8) = 187.1; P < 0.0001, respectively). Representative fluorescent images are presented corresponding to the image analysis of either sham treated with i.t. vehicle (Fig. 4K), CCI treated

with i.t. vehicle (Fig. 4L), or CCI treated with AM1241 (Fig. 4M). It is possible Inhibitors,research,lifescience,medical that overall changes in spinal cord cell numbers could dramatically alter dorsal horn immunofluorescent intensity quantification, as proliferation of Metalloexopeptidase microglia (Suter et al. 2009), astrocytes (Tsuda et al. 2011), or leukocyte CNS extravasation (Xu et al. 2007) have been reported. Consequently, cells could simply be constitutively expressing low levels of proteins, thus diminishing interpretation that a protein-specific cellular response has occurred following either CCI and/or i.t. AM1241. However, we observed no change in cell numbers as assessed by quantification of nuclear-specific DAPI fluorescence intensity as a consequence of either CCI procedures (ANOVA, F(1,8) = 0.1076; P = 0.7514 and ANOVA, F(1,8) = 0.7780; P = 0.4035, respectively) or i.t. drug injections (ANOVA, F(1,8) = 0.04328; P = 0.8404 and ANOVA, F(1,8) = 0.06960; P = 0.7986, respectively) (Fig. 4I and 4J).