Figure 1.
Effect of SNP on cGMP accumulation and cell migration in murine and human aortic VSMCs.
(A) cGMP accumulation was evaluated after treatment with a vehicle or SNP (40 µmol/L) for 5 minutes in murine VSMCs. n = 6 to 8 in each group. (B) Murine VSMCs migration was evaluated using a modified Boyden chamber assay after 24 hours of incubation with a vehicle or SNP (40 µmol/L) and/or Ang II (40 µmol/L). n = 12 to 13 in each group. (C) cGMP accumulation was evaluated after treatment with a vehicle or SNP (40 µmol/L) for 5 minutes in human aortic VSMCs. n = 5 to 7 in each group. (D) Human aortic VSMCs migration was evaluated using a modified Boyden chamber assay after 24 hours of incubation with a vehicle or SNP (40 µmol/L) and/or Ang II (40 µmol/L). n = 12 in each group. *P<0.05 compared to control; §P<0.0001 compared to control; §§P<0.0001 compared to Ang II. **P<0.01 compared to control.
Figure 2.
Effect of SNP on Ang II-induced ROCK activation in murine VSMCs.
Murine VSMCs were treated with a vehicle or SNP (40 µmol/L) for 3 hours and stimulated with saline or Ang II (40 µmol/L) for the last 1 hour. (A) Representative results of Western blot analysis for p-MBS, t-MBS, ROCK1, ROCK2, and actin. (B) Quantitative analysis of relative ROCK activity (p-MBS/t-MBS). (C) Quantitative analysis of relative ROCK1 expression. (D) Quantitative analysis of relative ROCK2 expression. n = 3 in each group. **P<0.01 compared to control; ††P<0.01 compared to Ang II; *P<0.05 compared to control; †P<0.05 compared to Ang II.
Figure 3.
Effect of SNP on Ang II-induced ROCK activation in human aortic VSMCs.
Human aortic VSMCs were treated with a vehicle or SNP (40 µmol/L) for 3 hours and stimulated with saline or Ang II (40 µmol/L) for the last 1 hour. (A) Representative results of Western blot analysis for p-MBS, t-MBS, ROCK1, ROCK2, and actin. (B) Quantitative analysis of relative ROCK activity (p-MBS/t-MBS). (C) Quantitative analysis of relative ROCK1 expression. (D) Quantitative analysis of relative ROCK2 expression. n = 4 in each group. *P<0.05 compared to control; ††P<0.01 compared to Ang II. †P<0.05 compared to Ang II.
Figure 4.
Effect of SNP on Ang II-induced ROCK activation in mouse aortas.
Mice were treated with saline or Ang II via implanted micro-osmotic pump infusion (2 mg/kg/day) and treated with saline or SNP via subcutaneous injection (5 mg/kg once a day) for 14 days. Increased blood pressures in Ang II-treated mice were normalized to levels similar to those in mice without Ang II treatment by hydralazine included in drinking water (20 mg/kg/day). (A) Representative results of Western blot analysis for p-MBS, t-MBS, ROCK1, ROCK2, and actin. (B) Quantitative analysis of relative ROCK activity (p-MBS/t-MBS). (C) Quantitative analysis of relative ROCK1 expression. (D) Quantitative analysis of relative ROCK2 expression. n = 5–11 in each group; **P<0.01 compared to control; ††P<0.01 compared to Ang II.
Table 1.
Body Weight, Systemic Hemodynamics and Heart Weight/Tibial Length in Control, Ang II-infused, SNP-infused, and A combined Ang II and SNP-infused Mice.