Fig. 7

PM regulates glycer-AGE-mediated cellular signaling for attracting endothelial dysfunction. The HUVECs were co-treated with various concentrations of PM (10 μM) and NAC (1000 μM) including with glycer-AGEs (100 μg/mL) for the indicated treatment times. The specific inhibitors (10 μM) were pre-treated before the glycer-AGE stimulation in all experiments, specifically: PD98059; the ERK inhibitor, SP600125; the JNK inhibitor, SB203580; the p38 inhibitor, BAY11-7082; and the NF-κB inhibitor. Relative expression of control was analyzed using Image J. a The HUVECs were co-treated with PM (10 μM) and NAC (1000 μM) including glycer-AGEs (100 μg/mL) for 4 h, and then the cells were collected with PBS for nucleic fraction and analyzed with 10% SDS-PAGE with PCNA as a control. Relative quantification of the bands was analyzed using Image J software. b The HUVECs were co-treated with PM (10 μM) and NAC (1000 μM) including glycer-AGEs (100 μg/mL) for 24 h, and then the THP-1 cells were co-cultured on the HUVECs for 1 h in a dark CO₂ incubator. Before the co-culture, the THP-1 cells were pre-labeled by BCECF-AM (100 μM) for 30 min. After the co-culture periods, the un-attached monocytes were washed twice with PBS. BCECF-AM-labeled monocytes were detected using confocal microscopy (100× magnification). c The cells were treated with the same method in Fig. 7b. After the co-culture periods, cell lysis by 0.1% SDS in 50 mM Tris-buffer (pH 7.0) was detected using a fluorescence multi-plate reader in excitation 485 nm and emission 535 nm. The fluorescence was quantified and analyzed with Duncan’s multiple range test as means ± SD at p < 0.05