Effects of vitamin D-induced supernatant of placental explants from preeclamptic women on oxidative stress and nitric oxide bioavailability in human umbilical vein endothelial cells

Figures

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  Figure 1. Levels of vitamin 25 (OH) D in plasma (A) and vitamin D receptor (VDR) gene expression (B) in the placental explants from preeclamptic (PE) (n=8) and normotensive (NT) pregnant women (n=8). Data are reported as means±SD. *P<0.05 (Mann-Whitney U test). a.u.: arbitrary units.

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  Figure 2. Concentration of cytokines in supernatant of placental explants from preeclamptic (PE) (n=8) and normotensive (NT) pregnant women (n=8) under different treatments. Data are reported as median with range. *P<0.05 (Mann-Whitney U test). IL-1β: interleukin-1 beta; TNF-α: tumor necrosis factor-alpha; IL-18: interleukin-18; VD: vitamin D.

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  Figure 3. Levels of nitrite (A) and nitrate (B) in the supernatant of human umbilical vein endothelial cells treated with supernatant of placental explants from preeclamptic (PE) (n=8) and normotensive (NT) pregnant women (n=8), cultured in the absence (Control) or presence of H2O2, H2O2+VD, and VD. Data are reported as means±SD for one independent experiment, performed in quintuplicates. *P<0.05 (ANOVA followed by Bonferroni's multiple comparison test). VD: vitamin D.

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  Figure 4. Nitric oxide fluorescence signal measured using DAF-FM™ after human umbilical vein endothelial cells (HUVEC) were exposed to placental explant supernatants from preeclamptic (PE) (n=8) and normotensive (NT) pregnant women (n=8). A, Control, Control NT, and Control PE; B, effect of treatment with vitamin D (VD) at 60 min; C, effect of treatment with H2O2 at 60 min; and D, effect of treatment with H2O2+VD at 60 min. Data are reported as means±SD for one independent experiment, performed in quintuplicates. *P<0.05 (ANOVA followed by Bonferroni's multiple comparison test). a.u.: arbitrary units.

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  Figure 5. Malondialdehyde (MDA) levels (A) and levels of intracellular reactive oxygen species (ROS) (B) evaluated by fluorescence of 2,7-dichlorodihydrofluorescein diacetate (DCFH-DA) in the supernatant of human umbilical vein endothelial cells (HUVEC) treated with supernatant of placental explants from preeclamptic (PE) (n=8) and normotensive (NT) pregnant women (n=8), cultured in the absence (Control) or presence of H2O2, H2O2+vitamin D (VD), and VD. Data are reported as means±SD. *P<0.05 (ANOVA followed by Bonferroni's multiple comparison test).