Butyrate sensitizes the release of substance P and calcitonin gene-related peptide evoked by capsaicin from primary cultured rat dorsal root ganglion neurons

To investigate whether butyrate increases substance P (SP) and calcitonin gene-related peptide (CGRP) release evoked by capsaicin from primary cultured dorsal root ganglion (DRG) neurons. DRG was dissected out from embryonic 15-day-old Wistar rat and cultured as dissociate cells for 24 h then exposed to butyrate (0.01 mmol/L, 0.1 mmol/L, 1 mmol/L, 10 mmol/L, respectively) for another 48 h. The neurons cultured continuously in media served as normal control. All above cultured samples were processed for detecting expression of mRNA for SP, CGRP and vanilloid receptor 1 (VR1) of DRG neurons by RT-PCR, and VR1 protein expression by Western blot. SP and CGRP basal release levels were measured by radioimmunoassay (RIA). After that, the DRG cells for RIA were stimulated by capsaicin (300 nmol/L) for 5 min and the culture media were harvested for detecting SP and CGRP release levels by RIA. The neurons exposed to vehicle solution served as vehicle controls. Exposure of butyrate on DRG neurons at higher concentrations (1 mmol/L, 10 mmol/L) for 48 h increased expression mRNA for SP and CGRP than that at lower concentrations (0.01 mmol/L, 0.1 mmol/L) and normal control (P