Fig. 5From: Advanced glycation end-products decreases expression of endothelial nitric oxide synthase through oxidative stress in human coronary artery endothelial cellsEffects of AGEs, SeMet and MnTBAP on activities of NOX, CAT, and SOD and eNOS mRNA levels in HCAECs. a Effect of SeMet on NADPH oxidase (NOX) activity. Cells were treated with AGEs (100 μg/ml) for 24 h, and NOX activities were determined by lucigenin-enhanced chemiluminescence with the presence of its substrate-NADPH. O2 − scavenger Tiron or flavoprotein inhibitor DPI was included in the assay to confirm the specificity of NOX activity. b Effect of SeMet on CAT activity. Cells were treated with AGEs (100 μg/ml) and/or antioxidant SeMet (20 μM) for 24 h and CAT activity was determined with acommercial kit. c Effect of SeMet on SOD activity. Cells were treated with AGEs (100 μg/ml) and/or antioxidant SeMet (20 μM) for 24 h and SOD activity was determined with acommercial kit. d Effect of SeMet on eNOS mRNA levels. Cells were treated with AGEs (100 μg/ml) and/or antioxidant SeMet (20 μM) for 24 h, and eNOS mRNA levels were determined by real-time PCR analysis. e Effect of MnTBAP on eNOS mRNA levels. Cells were treated with AGEs (100 μg/ml) and/or MnTBAP (2 μM) for 24 h, and eNOS mRNA levels were determined by real-time PCR analysis. f Effect of MnTBAP on eNOS protein levels. Cells were treated with AGEs (100 μg/ml) and/or MnTBAP (2 μM) for 24 h, and eNOS protein were determined by western blot. Full-length blots are presented. * P < 0.05, compare with control, # P < 0.05, compare with AGEs treatment, n = 3. Data are means and SE of multiple experiments (n)Back to article page