Exogenous soluble VEGF receptor-1 (sFlt-1) regulates Th1/Th2 cytokine production from normal placental explants via intracellular calcium

Hypertens Pregnancy. 2009 Aug;28(4):448-56. doi: 10.3109/10641950902777721.

Abstract

Objective: The increase of soluble VEGF-Receptor 1 (sFlt-1) is thought to contribute to the pathogenesis of preeclampsia. Soluble VEGF-Receptor 1 binds to circulating free VEGF and PLGF and this cascade is associated with endothelial dysfunction, a prominent feature of preeclampsia. Preeclampsia is also associated with excessive maternal response to pro-inflammatory stimuli manifesting as an imbalance of Th1/Th2 cytokine production at the maternal-fetal interface. Whether increased sFlt-1 expression has any effect on placental production of Th1/Th2 cytokines IL-10 and TNF-alpha is yet to be investigated. The aim of this study is to examine if exogenous sFlt-1 can regulate Th1/Th2 cytokines IL-10 and TNF-alpha production from normal placental explants via intracellular calcium release.

Methods: Placental explants were taken from the decidual surface of normal non-laboured term placentas (n = 11).Villous explants were cultured with increasing concentrations of sFlt-1. The dose effect of sFlt-1 on placental Th1 and Th2 cytokine production (TNF-alpha and IL-10) were examined. Free PLGF, VEGF and sFlt-1 concentrations in the conditioned medium were also measured. Intracellular calcium blocker, 1,2-bis-(o-aminophenoxy)-ethane-N,N,N',N'-tetraacetic acid, tetra(acetoxymethyl)-ester (BAPTA/AM) was applied to investigate whether the changes in cytokine concentration were mediated by intracellular free calcium.

Results: Placental IL-10 and TNF-alpha production were significantly increased after sFlt-1 incubation. The increase in IL-10 can be inhibited by BAPTA/AM. Soluble Flt-1 and free PLGF concentration in the conditioned medium was not changed. Free VEGF concentration in the conditioned medium was not detectable.

Conclusion: Exogenous sFlt-1 can increase TNF-alpha and IL-10 production from normal placental explants. The change in Th1/Th2 cytokine level may be mediated by intracellular free calcium.

MeSH terms

  • Calcium / metabolism*
  • Dose-Response Relationship, Drug
  • Egtazic Acid / analogs & derivatives
  • Egtazic Acid / pharmacology
  • Enzyme-Linked Immunosorbent Assay
  • Female
  • Growth Hormone / metabolism
  • Humans
  • Interleukin-10 / biosynthesis*
  • Organ Culture Techniques
  • Placenta / drug effects
  • Placenta / metabolism*
  • Placental Hormones / metabolism
  • Pregnancy
  • Protein Isoforms / administration & dosage
  • Protein Isoforms / metabolism
  • Statistics, Nonparametric
  • Th1 Cells / metabolism*
  • Th2 Cells / metabolism*
  • Tumor Necrosis Factor-alpha / biosynthesis*
  • Vascular Endothelial Growth Factor Receptor-1 / administration & dosage
  • Vascular Endothelial Growth Factor Receptor-1 / metabolism*

Substances

  • GH2 protein, human
  • Placental Hormones
  • Protein Isoforms
  • Tumor Necrosis Factor-alpha
  • Interleukin-10
  • 1,2-bis(2-aminophenoxy)ethane N,N,N',N'-tetraacetic acid acetoxymethyl ester
  • Egtazic Acid
  • Growth Hormone
  • Vascular Endothelial Growth Factor Receptor-1
  • Calcium