Fig. 4

Neutrophil elastase (NE) inhibition reduces the expression and production of inflammatory mediators (cytokines and chemokines) after spinal cord injury (SCI). Samples from sham or injured untreated (Injury) or after sivelestat treatment were prepared 1 day after injury as described in the Methods section. Representative section of tumor necrosis factor-alpha [TNF-α (a)], Interleukin [IL-6 (b)] immunofluorescence 1 day after SCI [3 fields/slide, n = 2–3/group (sham = 2, injury = 3 and sivelestat = 3)]. Total RNA from sham, vehicle (injury)- or sivelestat-treated samples were prepared DPI-1 after injury as described in the Methods section. RT-PCR results showing relative expression levels of TNF-α (c), IL-6 (d), inducible nitric oxide synthase [iNOS (e)], IL-1β (f), IL-10 (g), chemokine (C-C motif) ligand 2 [CCL-2 (h)], chemokine (C-C motif) ligand 3 [CCL-3 (i)] and TGF-β (j) after injury/treatment [n = 2–3/group (sham = 2, injury = 3 and sivelestat = 3) performed in triplicates]. GAPDH was used as internal controls for real-time quantitative reverse transcription–polymerase chain reaction. Data represent means ± S.E.M. ^##p < 0.01, ^###p < 0.001 vs. sham group. *p < 0.05, **p < 0.01, ***p < 0.001 vs. Injury group