TY - JOUR
T1 - Glucocorticoids inhibit angiogenesis and dysregulate angiogenic factors in the human placenta
AU - Stratta, Emily
AU - Palin, Victoria
AU - Sibley, Colin
AU - Brownbill, Paul
AU - Bischof, Helen
AU - Jones, Sarah
AU - Jones, Rebecca
PY - 2014/9
Y1 - 2014/9
N2 - Objectives: Fetal growth restriction (FGR) has been linked with excessive exposure to glucocorticoids (GCs) in animal and human pregnancies. In rodents, GC treatment reduces placental angiogenesis and inhibits expression of vascular endothelial growth factor (VEGF). We hypothesized that GCs inhibit angiogenesis in human placenta by dysregulating angiogenic factors.
Methods: Human umbilical vein endothelial cells (HUVECs, n=3) and primary human placental arterial endothelial cells (HPAECs, n=7) were treated with hydrocortisone, prednisolone and dexamethasone (10-1,000nM). Tube-like structure (TLS) formation in matrigel and cellular migration following scratching were assessed after 24 and 48h respectively. Cell viability, proliferation and apoptosis were assessed by MTT, APO% and LDH assays. Additional angiogenesis assays were conducted in the presence of angiogeneic factors (VEGF-A, FGF-2 and IL-8; 10nM). Chorionic plate arteries (CPA) from term placentas (n=10) and first trimester villous tissue were cultured for 48h with GCs (1000nM). mRNA expression of angiogenic regulators was quantified by Q-PCR.
Results: All three GCs dose-dependently reduced TLS formation in HUVECs and HPAECs (to 20-30% of vehicle, p<0.001). Cell migration was also significantly reduced (to 40-50% of vehicle, p<0.05) by 1000nM GCs only. Cell proliferation, apoptosis and viability were unaffected. GC treatment reduced CPA and villous tissue expression of angiogenic factors: fibroblast growth factor-2 (p<0.001), interleukin-8 (p<0.001), VEGF-A (p<0.01), VEGF-C (p<0.01), MMP-16 (p<0.01), MMP-1 (p<0.05) and CCL-2 (p<0.05). Co-treatment of HUVECs with VEGF-A, FGF-2 and IL-8 blunted the inhibitory effects on TLS formation and rescued the inhibitory effects on cell migration (p<0.05).
Conclusion: GCs treatment reduced key facets of angiogenesis in human placental endothelial cell models; GCs could therefore inhibit human placental angiogenesis and contribute to abnormal placental vascular development and the pathogenesis of FGR. The downregulation of specific angiogenic factors by GCs and the observed rescue effect of VEGF-A, FGF-2 and IL-8 identify putative mechanistic pathways involved.
AB - Objectives: Fetal growth restriction (FGR) has been linked with excessive exposure to glucocorticoids (GCs) in animal and human pregnancies. In rodents, GC treatment reduces placental angiogenesis and inhibits expression of vascular endothelial growth factor (VEGF). We hypothesized that GCs inhibit angiogenesis in human placenta by dysregulating angiogenic factors.
Methods: Human umbilical vein endothelial cells (HUVECs, n=3) and primary human placental arterial endothelial cells (HPAECs, n=7) were treated with hydrocortisone, prednisolone and dexamethasone (10-1,000nM). Tube-like structure (TLS) formation in matrigel and cellular migration following scratching were assessed after 24 and 48h respectively. Cell viability, proliferation and apoptosis were assessed by MTT, APO% and LDH assays. Additional angiogenesis assays were conducted in the presence of angiogeneic factors (VEGF-A, FGF-2 and IL-8; 10nM). Chorionic plate arteries (CPA) from term placentas (n=10) and first trimester villous tissue were cultured for 48h with GCs (1000nM). mRNA expression of angiogenic regulators was quantified by Q-PCR.
Results: All three GCs dose-dependently reduced TLS formation in HUVECs and HPAECs (to 20-30% of vehicle, p<0.001). Cell migration was also significantly reduced (to 40-50% of vehicle, p<0.05) by 1000nM GCs only. Cell proliferation, apoptosis and viability were unaffected. GC treatment reduced CPA and villous tissue expression of angiogenic factors: fibroblast growth factor-2 (p<0.001), interleukin-8 (p<0.001), VEGF-A (p<0.01), VEGF-C (p<0.01), MMP-16 (p<0.01), MMP-1 (p<0.05) and CCL-2 (p<0.05). Co-treatment of HUVECs with VEGF-A, FGF-2 and IL-8 blunted the inhibitory effects on TLS formation and rescued the inhibitory effects on cell migration (p<0.05).
Conclusion: GCs treatment reduced key facets of angiogenesis in human placental endothelial cell models; GCs could therefore inhibit human placental angiogenesis and contribute to abnormal placental vascular development and the pathogenesis of FGR. The downregulation of specific angiogenic factors by GCs and the observed rescue effect of VEGF-A, FGF-2 and IL-8 identify putative mechanistic pathways involved.
U2 - 10.1016/j.placenta.2014.06.055
DO - 10.1016/j.placenta.2014.06.055
M3 - Article
SN - 0143-4004
VL - 35
SP - A16
JO - Placenta
JF - Placenta
IS - 9
ER -