Figure 5.

Inhibition of AR prevents HG- and/or TNF-α-induced phosphorylation of cdk-2 and c-myc expression and ROS production in VSMC. A and B, Quiescent VSMC (n = 5 for each treatment) were preincubated with 10 μm sorbinil or carrier for 24 h followed by stimulation with HG or TNF-α for different time intervals (A) and 24 h (B). The pooled total cell lysate was subjected to Western blot analysis using antibodies against phospho-cdk-2, total cdk-2, and c-myc. C, Growth-arrested VSMC were preincubated with sorbinil or carrier for 24 h followed by stimulation with HG and/or TNF-α for 4 h to determine c-myc mRNA expression by RT-PCR. D, For the determination of ROS, growth-arrested VSMC were preincubated with sorbinil or carrier for 24 h followed by stimulation with HG and/or TNF-α for 1 h, and the cells were treated with dichlorodihydrofluorescein for 15 min, washed, and fluorescence determined using excitation and emission wavelengths of 485 and 528 nm, respectively. Densitometric analysis was performed using Kodak 1D image analysis software. Bars, Means ± sem of three independent analyses; #, P < 0.001 compared with control cells; *, P < 0.001; **, P < 0.05 compared with cells treated with HG and/or TNF-α.