Pre-Clinical Investigation of Cardioprotective Beta-Blockers as a Therapeutic Strategy for Preeclampsia

Figures

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  Figure 1. Beta-blockers had limited effect on secreted anti-angiogenic factors across different tissue types. Relative sFlt-1 secretion in HUVCEs (A) was significantly reduced with carvedilol (Carv) treatment, but not bisoprolol (Bis) treatment or metoprolol (Met) treatment. Relative sFlt-1 secretion was unchanged in trophoblasts (B) and explants (C) following beta-blocker treatment with Carv, Bis and Met. In HUVEC, relative sENG secretion (D) was decreased with treatment at the top dose of Met, but unchanged with Carv and Bis treatment. In explants, relative sENG secretion (E) was decreased with treatment at 100 uM Bis, but unchanged at the other doses or with Carv or Met. Data are mean ± SEM, expressed relative to control, n = 3–4, ** p < 0.01, **** p < 0.0001.

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  Figure 2. Endoglin mRNA is upregulated in HUVECs following beta-blocker treatment. Placental explant expression of sFlt-1 isoforms, sFlt-1-e15a (A) and sFlt-1-i13 (B) was not affected by treatment with beta-blockers carvedilol (Carv), bisoprolol (Bis), or metoprolol (Met). HUVEC expression of ENG mRNA (C) was significantly increased with top dose Carv and Met, but not Bis. Data are mean ± SEM, expressed as a percentage of control, n = 3–4, ** p < 0.01, *** p < 0.001.

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  Figure 3. Pro-angiogenic factors upregulated following beta-blocker treatment of HUVECs, but not explants. VEGF mRNA expression in HUVECs (A) was significantly increased with metoprolol (Met) treatment, but not carvedilol (Carv) or bisoprolol (Bis). PGF mRNA expression in HUVECs (C) was significantly increased with the top dose of Carv, Bis, and Met. ADM mRNA expression in HUVECs (E) was significantly increased with top dose Carv treatment and Met treatment, but not Bis. VEGF and PGF mRNA expression were unchanged in placental explants following beta-blocker treatment (B,D, respectively). Data are mean ± SEM, expressed as a percentage of control, n = 3–4, * p < 0.05, ** p < 0.01, **** p < 0.0001.

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  Figure 4. Inflammatory markers are altered following beta-blocker treatment. Expression of IL-1b mRNA in HUVECs (A) was not affected by treatment with beta-blockers carvedilol (Carv), bisoprolol (Bis), or metoprolol (Met). When TNFα is added to HUVECs to induce endothelial dysfunction, IL-1b expression is significantly increased from control levels (B) and decreases with doses of Bis and Met, but not Carv. TNFα induced endothelial dysfunction also significantly increases mRNA levels of IL-6 (C) and NLRP3 (D). Beta-blocker treatment with top dose Carv, Bis, and Met significantly decreases induced IL-6 expression (C). NLRP3 mRNA expression was not affected by beta-blockers treatment (D). Expression of PTGS2 mRNA in HUVECs (E) was significantly increased following top dose beta-blocker treatment with Carv, Bis, and Met. PTGS2 expression is significantly induced with TNFα (endothelial dysfunction) (F), which is further increased with top dose Carv, decreased with 100 uM Bis, and unaffected by Met. Data are mean ± SEM, expressed as a percentage of control (HUVEC) or TNFα control (endothelial dysfunction), n = 3–4, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

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  Figure 5. Antioxidant HO-1 is increased with beta-blocker treatment. Expression of HO-1 mRNA is significantly increased in HUVECs following top dose beta-blocker treatment with carvedilol (Carv), bisoprolol (Bis), and metoprolol (Met). Data are mean ± SEM, expressed as a percentage of control, n = 3–4, *** p < 0.001, **** p < 0.0001.

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  Figure 6. In a model of endothelial dysfunction, VCAM mRNA expression is decreased with beta-blocker treatment. In HUVECs treated with TNFα to induce endothelial dysfunction (A), VCAM mRNA is significantly reduced with top dose beta-blockers, carvedilol (Carv), bisoprolol (Bis), and metoprolol (Met). In the same model, ET-1 mRNA (B) and ETB mRNA (C) expression are not affected by Carv, Bis, or Met. Data are mean ± SEM, expressed as a percentage of TNFα control, n = 3–4, * p < 0.05, ** p < 0.01, **** p < 0.0001.