Figure 4. Augmentation of the cGMP pathway improves vascular outcomes in the sFlt1 overexpression model.

(A) Pregnant mice given sFlt1 adenovirus at gd8 (arrow) with sildenafil citrate (sFlt1 + SILD, n = 5) maintained normal blood pressure, whereas untreated mice developed hypertension (n = 3). *P < 0.001, sFlt1 mice compared with earlier gestation days. Two-way repeated-measures ANOVA. Data for sFlt1 mice in this figure were obtained from data presented as sFlt1 group in Figure 1C. (B) Comparison of MAP and plasma sFLT1 levels between sFlt1 (n = 3) and sFlt1 + SILD mice (n = 5). ΔMAP was obtained by subtraction of individual pressure at gd17 from that at gd7. Gd7 was selected as the latest time point when all animals had no pressure differences (prior to intervention, permitting comparison). ΔMAP was significantly lower in sFlt1 + SILD–treated mice (*P < 0.01), while plasma sFLT1 levels at gd17 were similar. Data for sFlt1 mice in this figure were obtained from data presented as sFlt1 group in Figure 1D. (C) Ex vivo contraction of mesenteric vessels in response to Ang II was restored by treatment with oral sildenafil in comparison with sFlt1 mice at gd17 (n = 8 per group). *P < 0.05. (D) Representative images of aorta isolated from sFlt1 (left) and sFlt1 + SILD (right) mice at gd17 staining for MitoSOX Red (top; scale bar: 50 μm) and nitrotyrosine (bottom; scale bar: 20 μm). (E) Quantitation of MitoSOX Red immunofluorescence in aortic tissue from gd17 sFlt1- and sFlt1 + SILD–treated mice. OD per area (pixels²) of tissue was calculated in 5 high-power fields per sample (n = 3 aortas per group). *P < 0.001 versus sFlt1 by 2-way ANOVA and Tukey’s post hoc test. Data for sFlt1 mice in this figure were obtained from summary sFlt1 data presented in Supplemental Figure 3B. (F) Sildenafil treatment increased urinary cGMP compared with sFlt1 alone. (n = 7 per group.) *P < 0.01. Data represent the mean ± SEM, analyzed by unpaired 2-tailed t test with Welch’s correction (except where noted).