Evaluating Anti-Inflammatory Activity of TAP Extract

The in vitro anti-inflammatory activity of the TAP extract was evaluated by using a cell model previously described [27 (link)]. Briefly, R3/1 NF-κB cells (5000 cells/well) were pre-treated for 18 h with different concentrations of the extract (1–250 µg/mL) in complete medium (DMEM 10% FBS, 1% l-glutamine, 1% Penicillin/Streptomycin). Rosiglitazone 10 µM was used as a positive control [27 (link)]. Then, cells were stimulated for 6 h with 10 ng/mL IL-1α, and for 6 and 24 h with 10 ng/mL TNF-α. Luciferase measurements was performed with a luminometer (Wallac Victor2 1420, Perkin-Elmer™ Life Science, Monza, Italy) after adding 100 µL of ONE-Glo™ Luciferase Assay Substrate (purchased from Promega Corporation, Madison, WI, USA). Experiments were carried out with biological and technical replicates; values are reported as mean ± SD compared to untreated control cells. One-way ANOVA with Bonferroni’s multiple comparisons test (p < 0.05 was considered significant) was used for the statistical analysis.