Synergistic regulation of uterine radial artery adaptation to pregnancy by paracrine and hemodynamic factors

Figures

• 

  Figure 1.. Illustration of computational model of the human uteroplacental artery network. Schematic showing the modeled physiological scenarios at each time point (left) and the model perturbations (right) that were used for each time point, respectively. AV image was created with a licensed version of BioRender.com.

• 

  Figure 2.. Expression of vascular endothelial growth factor receptor (VEGFR)-1, VEGFR-2, estrogen receptor-α (ERα), and progesterone receptor (PR) in rat radial arteries. Representative images of immunohistochemical staining of nonpregnant (A–D) or pregnant (E–H) rat radial arteries stained for VEGFR-1 (A and E), VEGFR-2 (B and F), ERα (C and G), or PR (D and H). Rat placental tissue was used as positive controls (I–L), with irrelevant IgG on radial arteries used as negative controls (I–L, insets). Scale bars = 100 µm. Violin plots show quantification of the staining intensity of VEGFR-1 (M), VEGFR-2 (N), ERα (O), and PR (P) within all three layers (tunica intima, media, adventitia) of the radial artery walls of nonpregnant (NP) and pregnant (P) animals. n = 3 or 4/group; Mann–Whitney U test. *P < 0.05.

• 

  Figure 3.. Rat radial artery response to increasing flow rates in pressure myography. A–E: graphs showing the change of inner diameter normalized to size before onset of flow across the intramural pressure gradient range (PΔ) as acquired in myography experiments. Data of arteries from nonpregnant control animals (n = 8, pink) and from pregnant animals (n = 10, gray) are taken from Allerkamp et al. (1). Blue lines show treatment groups acquired in this work (nonpregnant radial arteries treated with a single paracrine factor, or combination of all 4; n = 5 or 6 per condition). Means ± SE. Two-way ANOVA + Tukey’s multiple comparison post hoc test. *P < 0.05. F: violin plots showing inner diameter (normalized to size before onset of flow) of all groups at 40-mmHg pressure drop. Dotted line shows mean size of untreated nonpregnant radial arteries. NP, untreated nonpregnant control; P, pregnant control; β-est, β-estradiol; Prog, progesterone; PlGF, placental growth factor; VEGF, vascular endothelial growth factor; Mix, combined factors. One-way ANOVA + Sidak’s multiple comparison post hoc test; n = 5–10/group. *P < 0.05, **P < 0.01, and ***P < 0.001.

• 

  Figure 4.. Influence of endothelial nitric oxide synthase (eNOS) on rat radial artery response to increasing flow rates after treatment with paracrine factors. A–D: graphs showing the change of inner diameter normalized to size before onset of flow across the intramural pressure drop range as acquired in myography experiments using nonpregnant (n = 8, gray) and pregnant (n = 10, pink) control arteries [data taken from Allerkamp et al. (1)], nonpregnant arteries treated with single/combined paracrine factors (n = 5 or 6, dark blue), or nonpregnant arteries treated with Nω-nitro-l-arginine methyl ester (l-NAME) before single/combined paracrine factors (n = 5 or 6, light blue). Means ± SE; two-way ANOVA + Tukey’s multiple comparison post hoc test. a = P < 0.05 for nonpregnant vs. pregnant control; b = P < 0.05 for nonpregnant control vs. nonpregnant + treatment + Nω-nitro-l-arginine methyl ester (l-NAME). E: violin plots showing inner diameter (normalized to size before onset of flow) of all groups at 40-mmHg pressure drop. Dotted line shows mean size of nonpregnant control group. NP, untreated nonpregnant control; β-est, β-estradiol; Prog, progesterone; PlGF, placental growth factor; Mix,…

• 

  Figure 5.. Model predictions of change of shear stress within human radial arteries in different scenarios. A–C: relationship between radial artery radius and shear stress in a plugged (solid line) or unplugged (dotted line) scenario at 6–8 (A), 10–12 (B), and 16–20 (C) wk of pregnancy. The cross and dot mark the physiological radial artery radius in each case. D: schematic of the model perturbations to reflect pathological scenarios. Removing the plug from the spiral arteries is labeled with process 1, reducing radial artery radius to 85% with process 2, and the combined effects with process 3. E: bar graphs showing the x-fold change of shear stress compared with the physiological scenario at 6–8, 10–12, and 16–20 wk of gestation when either 1) plugs are removed (scenario a), 2) radial artery diameter was reduced to 85% of its physiological value (scenario b), or 3) the combination of both (scenario c).

• 

  Figure 6.. Influence of paracrine factors on nitric oxide (NO) production in human microvascular endothelial-cell (HMEC)-1 cells under different shear stress. Graphs showing the relative fluorescence of HMEC-1 cells under shear stress of 7 (A), 12 (B), 16 (C), or 20 (D) dyn/cm2 over time for controls (Ctrl) or HMEC-1s treated with placental growth factor (PlGF), β-estradiol (β-est), progesterone (Prog), or the combination of all factors (Mix). Bar graphs showing the area under the curve (AUC) calculated for each condition are presented alongside. Data are from n = 3 or 4 biological replicates/group, run in technical duplicates. AUC was compared by one-way ANOVA + Dunnett’s post hoc test to each nontreated control group. Symbols and bar graphs show means ± SE. *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001.