Fig. 2From: Advanced glycation end-products decreases expression of endothelial nitric oxide synthase through oxidative stress in human coronary artery endothelial cellsEffects of AGEs on eNOS mRNA in HCAECs. HCAECs were cultured with different concentrations of AGEs for different periods of time. The mRNA levels of eNOS and glyceraldehyde-3-phosphatede-hydrogenase (GAPDH) were determined by real-time PCR analysis. a Concentration-dependent study. Cells were treated with different concentrations of AGEs (50, 100, or 200 μg/ml) for 24 h. b Time-dependent study. Cells were treated with AGEs (100 μg/ml) for different times (12, 24 and 48 h). c Effect of anti-RAGE antiboday. Cells were treated with 100 μg/ml AGEs and different concentrations of anti-RAGE antiboday for 30 min and followed with AGEs treatment for 24 h. Isotype IgG was used for a negative control. d eNOS mRNA stability. Cells were treated with 5 μg/ml actinomycin D in the presence or absence of AGEs (100 μg/ml) for indicated time points (0, 2, 4, 8, or 16 h), and eNOS mRNA levels were determined by real-time PCR. * P < 0.05, compare with control, # P < 0.05, compare with AGEs treatment, n = 3 experiments. Data are means and SE of multiple experiments (n)Back to article page