EPHB4, a down stream target of IFN-γ/STAT1 signal pathway, regulates endothelial activation possibly contributing to the development of preeclampsia

Problem: Preeclampsia is characterized by endothelial activation and excessive inflammation, of which interferon (IFN)-γ is a potent inducer. Eph receptor B4 (EPHB4) also involved in endothelial activation in inflammation. Their role and relationship in preeclampsia remain unclear.

Method of study: Intercellular adhesion molecular (ICAM)-1 was employed as the hallmark of endothelial activation. The serum levels of IFN-γ and the expression of EPHB4 and ICAM-1 were assessed by ELISA, qRT-PCR and WB, respectively. Primary human umbilical vein endothelial cells (HUVECs) were treated with IFN-γ of different concentration or for different times to determine the effect of IFN-γ on EPHB4 and ICAM-1 expression. Overexpression and shRNA constructs, chromatin immunoprecipitation (ChIP) and luciferase assays were conducted to clarify the regulation mechanism of IFN-γ/STAT1 on EPHB4 resulting in HUVECs activation. Endothelial-trophoblast co-culture model was used to illustrate the role of EPHB4 in the process of activated endothelial cells resisting trophoblast invasion.

Results: IFN-γ, EPHB4 and ICAM-1 expression were elevated in preeclampsia. IFN-γ induced HUVECs activation through EPHB4 expression. ChIP and luciferase assays revealed that IFN-γ promoted EPHB4 transcription by STAT-1 binding to EPHB4 promoter. EPHB4 probably involved in resisting trophoblasts displacement by IFN-γ-activated HUVECs.

Conclusion: This study uncovered the character of EPHB4-regulating endothelial activation in the pathogenesis of preeclampsia.