Figure 7. Functional role of Nox4 and Rho kinase in the regulation of vascular dysfunction by docetaxel in vivo.
(A) Schematic of the study design for WT mice injected with vehicle, docetaxel (10 mg/kg every 5 days), or docetaxel plus the Nox1/4 inhibitor GKT137831 (40 mg/kg every day) or placebo (solvent) for 3 weeks. (B) Systolic blood pressure by tail-cuff plethysmography (n = 6–7/group). Data are expressed as the mean ± SEM. #P < 0.01 versus baseline; **P < 0.01 versus WT (vehicle + placebo); ***P < 0.001 versus WT treated with docetaxel plus GKT137831; 2-way, repeated-measures ANOVA with Tukey’s test. (C) Endothelium-dependent vasorelaxation responses to ACh (1 nM–10 μM) and endothelium-independent vasorelaxation responses to SNP (1 nM–10 μM) in mouse aortas (n = 5–6/group). Data are expressed as the mean ± SEM. *P < 0.05 versus WT treated with docetaxel plus GKT137831; **P < 0.01 versus WT; 2-way, repeated-measures ANOVA with Tukey’s test. (D) Study design, for WT mice injected with docetaxel (10 mg/kg every 5 days) or docetaxel (10 mg/kg every 5 days) and the ROCK inhibitor fasudil (30 mg/kg every day) or placebo (solvent) for 3 weeks. (E) BP was measured by tail-cuff plethysmography (n = 6–7/group). Data are expressed as the mean ± SEM. #P < 0.01 versus baseline; ***P < 0.01 versus WT mice and WT mice treated with docetaxel plus fasudil; 2-way, repeated-measures ANOVA with Tukey’s test. (F) Endothelium-dependent vasorelaxation responses to ACh (1 nM–10 μM) and endothelium-independent vasorelaxation responses to SNP (1 nM–10 μM) in mouse aortas (n = 6/group). Data are expressed as the mean ± SEM. *P < 0.05 versus WT and WT mice treated with docetaxel plus fasudil; 2-way, repeated-measures ANOVA with Tukey’s test. (G) Effect of fasudil on phosphorylation of eNos at Thr495 in WT mice treated with docetaxel (n = 6/group). Densitometric analysis of proteins normalized to total eNos expression. Immunoblots are from 1 of 2 independent experiments. Data are expressed as the mean ± SEM. *P < 0.05 versus WT; **P < 0.01 versus WT mice treated with docetaxel plus fasudil; 1-way ANOVA with Tukey’s test.