Effects of Phytochemicals on Mast Cell Activation

1 × 10⁶ BMMCs/ml were cultured in triplicates with 10 ng/ml IL-3 and SCF. Cells were activated by pre-sensitizing with 1 µg/ml DNP-IgE (clone SPE7, Sigma) or vehicle (medium), followed by treatment with 200 ng/ml DNP-BSA (5 (link), 24 (link)). Increasing concentrations of curcumin (Sigma), bacitracin (Sigma), quercetin-3-rutinoside hydrate or rutin (Sigma), and propynoic acid carbamoyl methyl amide (PACMA)-31 (Sigma) or vehicle (DMSO) were added to various experimental groups for different time periods (including 30 min and 24 h) prior to challenge with DNP-BSA. Expression of cytokine genes was assessed by RT-PCR between 30 min to 1 h following activation. Assessment of secreted cytokines was performed 6 to 24 h later.