Fig. 7.

PHAR decreases inflammatory response in Raw264.7 cells stimulated with LPS. Low serum-depleted Raw264.7 cells were pre-treated to 10 μM of PHAR for 8 h. Then, cells were treated to 100 ng/ml of LPS for the indicated times. A, representative immunoblots of NRF2, HO1, p65, pre-IL1β, COX2, NOS2, and GAPDH as a loading control. The black arrowhead marks p65 and NOS2 specific bands. B–C, densitometric analysis of NRF2, pre-IL1b, COX2, and NOS2 protein levels from representative immunoblot from (A), expressed as a ratio of protein levels/GAPDH. Data are mean ± S.D. (n = 3). *p < 0.05; ***p < 0,001 vs vehicle or LPS according to a two-way ANOVA test. D-E, mRNA levels of Il1b, Ptgs2, Nos2, Il6, and Tnf were determined by qRT-PCR and normalized by Gapdh, Tbp, and Actb average. Data are mean ± S.D. (n = 4). *p < 0.05; ***p < 0.001 vs LPS according to a two-way ANOVA test.