Background: Spiral artery remodelling is crucial for a successful pregnancy. In a healthy human pregnancy, cells of the placenta called extravillous trophoblasts (EVT) invade into the decidua and interact with the endothelial cells (EC) and vascular smooth muscle cells (VSMC) of the maternal spiral arteries (SA) in a process known as spiral artery remodelling. This process results in the development of low-resistance, dilated SAs that increase the blood supply to the developing fetus. An in vitro model has shown several molecules to be upregulated in response to trophoblast conditioned media (TCM) stimulation, including MMP10, an enzyme involved in ECM breakdown, but its role in spiral artery remodelling has yet to be determined.
Aim: The aim of this project was to investigate the role of MMP10 in SA remodelling.
Methods: Experiments were carried out using the human endothelial line SGHEC-7, cultured in 2 ml 5% FCS phenyl-red media. The cells were then stimulated in 0% FCS phenyl red-free media with 100 ng/ml TCM or varying concentrations of IL1β or PMA and incubated for 48 h. ECs were also stimulated with TCM in the presence of an IL1β neutralising antibody. The supernatant was then collected and an R&D Systems human Total MMP-10 (catalogue number: DY910) DuoSet ELISA Development kit was used to detect MMP10. The cell monolayer for each experiment was then frozen and used for subsequent protein determination via Bradford Assay.
Results: TCM was shown to stimulate MMP10 secretion by ECs. MMP10 secreted by ECs increased significantly in a dose respondent manner when stimulated with 05 ng/ml of IL1β. Addition of an IL1β neutralising antibody to ECs stimulated with TCM decreased the amount of MMP-10 produced.
Conclusion: This data suggests that IL1β (within TCM) may be stimulating secretion of MMP10 by ECs.
11 Jul 2016 - 11 Jul 2016