Nitric oxide (NO) reversed TNF-α inhibition of trophoblast interaction with endothelial cellular networks

B Xu; F Charlton; A Makris; A Hennessy

doi:10.1016/j.placenta.2014.03.009

Nitric oxide (NO) reversed TNF-α inhibition of trophoblast interaction with endothelial cellular networks

Placenta. 2014 Jun;35(6):417-21. doi: 10.1016/j.placenta.2014.03.009. Epub 2014 Mar 19.

Authors

B Xu¹, F Charlton², A Makris³, A Hennessy⁴

Affiliations

¹ School of Medicine, University of Western Sydney, Sydney, Australia; Vascular Immunology Research Laboratory, The Heart Research Institute, University of Sydney, Sydney, Australia. Electronic address: bei_xu2001@yahoo.com.
² Vascular Immunology Research Laboratory, The Heart Research Institute, University of Sydney, Sydney, Australia.
³ Renal Unit, Liverpool Hospital, Sydney, Australia; Vascular Immunology Research Laboratory, The Heart Research Institute, University of Sydney, Sydney, Australia.
⁴ School of Medicine, University of Western Sydney, Sydney, Australia; Vascular Immunology Research Laboratory, The Heart Research Institute, University of Sydney, Sydney, Australia.

PMID: 24695240
DOI: 10.1016/j.placenta.2014.03.009

Abstract

Introduction: The interaction between trophoblast cells and maternal uterine endothelium is important for placental vascular modelling. Nitric oxide (NO) is a potent vasorelaxant that regulates systemic blood pressure and is reduced in preeclampsia. NO may affect placental cell interaction.

Objectives: This study was to examine whether NO plays a role in regulating TNF-α induced inhibition of trophoblast cell integration into endothelial cellular networks in-vitro.

Methods: Red fluorescent-labelled human uterine myometrial microvascular endothelial cells (UtMVECs) were seeded on Matrigel. After endothelial cellular networks formed, green fluorescent-labelled HTR-8/SVneo trophoblast cells were co-cultured with endothelial cells, together with/without TNF-α (0.5 ng/ml) and/or NO donor, sodium nitroprusside dihydrate (SNP) (100 μM). Images were captured after 24 h. The effects on HTR-8/SVneo cell integration were quantified by Image Analysis software. The cells were then recovered from Matrigel to extract mRNA. Quantitative PCR was performed to evaluate the expression of eNOS, VCAM-1 and E-selectin. The concentrations of sVCAM-1 and sE-selectin in the conditioned medium were measured by ELISA.

Results: TNF-α inhibited HTR-8/SVneo trophoblast cell integration into endothelial cellular networks, as well as decreased eNOS mRNA expression. Increases in VCAM-1 and E-selectin in cellular mRNA and protein concentrations in the conditioned medium were also seen. The NO donor reversed the inhibitory effect of TNF-α on trophoblast integration and increased eNOS mRNA expression. SNP also reduced sE-selectin and sVCAM-1 expressions which were increased by TNF-α in the conditioned medium.

Conclusion: Our data suggest the inhibitory effect of TNF-α on trophoblast integration may be mediated by NO, via reducing endothelial cell activation.

Keywords: E-selectin; Nitric oxide; TNF-α; Trophoblast–endothelial cell interaction; VCAM-1; eNOS.

MeSH terms

Cells, Cultured
Coculture Techniques
Culture Media, Conditioned / chemistry
E-Selectin / analysis
E-Selectin / genetics
Endothelial Cells / chemistry
Endothelial Cells / physiology*
Female
Fluorescent Dyes
Humans
Nitric Oxide / pharmacology*
Nitric Oxide Donors / pharmacology
Nitric Oxide Synthase Type III / analysis
Nitric Oxide Synthase Type III / genetics
Nitroprusside / pharmacology
Pregnancy
RNA, Messenger / analysis
Trophoblasts / chemistry
Trophoblasts / physiology*
Tumor Necrosis Factor-alpha / pharmacology*
Uterus / blood supply*
Uterus / cytology
Vascular Cell Adhesion Molecule-1 / analysis
Vascular Cell Adhesion Molecule-1 / genetics

Substances

Culture Media, Conditioned
E-Selectin
Fluorescent Dyes
Nitric Oxide Donors
RNA, Messenger
Tumor Necrosis Factor-alpha
Vascular Cell Adhesion Molecule-1
Nitroprusside
Nitric Oxide
Nitric Oxide Synthase Type III