Figure 6.

MAPK and NF-κB activation in dendritic cells (DCs). Five-day human DCs pretreated or not with resveratrol (Resv, 50 μM) were cultured for 45 minutes with or without AGE-albumin (AGE, 30 μg/mL). DCs treated with albumin (Alb; 30 μg/mL), resveratrol (Resv; 50 μM), LPS (0.2 μg/mL), or PMA (0.2 μg/mL) were used as controls. Cells were then analyzed by cell-based ELISA MAPK assay to monitor p38 and ERK activation and by NF-κB (p65 and p50) transcription factor assay to monitor NF-κB activation. (a) AGE-albumin stimulation induced the activation of both MAPK p38 and ERK pathways in DCs. Pretreatment of DCs with resveratrol prevented the upregulation of both MAPKs in response to AGE-albumin (n = 4; p-p38/p38: ^∗‡ P < 0.001, ^† P < 0.01; pERK/ERK: ^∗‡ P < 0.001, ^† P < 0.05). (b) AGE-albumin stimulation significantly increased active p65 and p50 levels in DCs. Pretreatment of DCs with resveratrol prevented the upregulation of both active p50 and p65 in response to AGE-albumin. The results are expressed as arbitrary units (n = 4, p50 and p65: *P < 0.05; ^† P < 0.001).