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Fig. 5 | Cardiovascular Diabetology

Fig. 5

From: Advanced glycation end-products decreases expression of endothelial nitric oxide synthase through oxidative stress in human coronary artery endothelial cells

Fig. 5

Effects 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)

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