Figure 6.

MaR1 reduces the number of leukocytes proximal to CGRP⁺ fibres and the release of CGRP by DRG neurons. Hind paw skin was dissected for determination of total leukocytes (CD11b⁺ cells) close to CGRP⁺ fibres (a), which showed an increase of total CD11b fluorescence proximal to CGRP⁺ fibres in the CFA group and reduction after MaR1 treatment. For CGRP release assay (b), naïve DRG neurons received vehicle or different concentrations of MaR1 (0.3, 1, or 3 ng·ml⁻¹) before stimulus with capsaicin. Supernatant was collected 1 hr after capsaicin to determine CGRP levels by EIA. Panels (c) to (e) analysed with further detail the cellular types recruited to the paw skin during CFA inflammation. LysM‐eGFP (C57BL/6 background mice) was used to determine neutrophils and macrophages (c). The staining neutrophils (GR‐1, d) and macrophages (CD68, e) in hind paw skin samples of Swiss mice also showed that MaR1 reduced CFA‐induced recruitment of these cells. Results are expressed as mean ± SEM, n = 6 mice per group per experiment, two independent experiments (*P < 0.05 vs. saline, ^# P < 0.05 vs. 0 mg·kg⁻¹ group; one‐way ANOVA followed by Tukey's post‐test). Results are expressed as mean ± SEM, n = 6 wells per group per experiment, two independent experiments (*P < 0.05 vs. saline, ^# P < 0.05 vs. vehicle group; ^** P < 0.05 vs. 1 ng·ml⁻¹ group; one‐way ANOVA followed by Tukey's post‐test)