Fig. 2

Effects 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)