Placenta-derived Chymotrypsin-like Protease (CLP) Disturbs Endothelial Junctional Structure in Preeclampsia

Abstract

Placenta-derived chymotrypsin-like protease may contribute to endothelial activation in preeclampsia. In this study, we determined if placenta-derived chymotrypsin-like protease could disturb endothelial junctional integrity to promote endothelial permeability in preeclampsia. Confluent endothelial cells were cocultured with placental trophoblasts or treated with preeclampsia placenta-conditioned medium. Endothelial junction protein vascular endothelial cadherin expression and distribution were examined by fluorescent staining of endothelial cells with or without depletion of chymotrypsin. The association of endothelial cell junction protein complex VE-cadherin/beta-catenin/p120 was examined by a combined immuno-precipitation and immuno-blotting assay. Our results showed that endothelial cells cocultured with preeclampsia trophoblasts or exposed to preeclampsia placental conditioned medium exhibited a discontinuous distribution and reduced expression of vascular endothelial cadherin at cell contact regions. Vascular endothelial cadherin and p120 were expressed in control endothelial cells, but reduced or lost in endothelial cells exposed to preeclampsia placental conditioned medium, suggesting that the junctional protein complex of VE-cadherin/beta-catenin/p120 was disrupted in endothelial cells exposed to preeclampsia placental conditioned medium. We also observed that removal of trophoblasts from the coculture system and depletion of the protease from the preeclampsia placental conditioned medium could restore the dysregulated endothelial junction protein expression and distribution. Chymotrypsin also induced a dose dependent increase in endothelial monolayer permeability. We conclude that chymotrypsin-like protease released by the placenta is at least one important mediator responsible for disrupting endothelial cell integrity and inducing endothelial permeability in preeclampsia.