Inhibition of the PI3K-Akt Pathway Suppresses sFlt1 Expression in Human Placental Hypoxia Models In Vitro

Abstract

Objective: Although elevated expression of soluble fms-like tyrosine kinase 1 (sFlt1) plays a major role in the pathogenesis of pre-eclampsia, it is unclear how hypoxia regulates placental sFlt1 expression. Thus, we investigated sFlt1 expression in placentas from normal and preeclamptic pregnancies and in human placental hypoxia models in vitro to examine the role of the PI3K-Akt pathway in regulating the expression of this molecule. Methods: We examined the expression of VEGF, PIGF, sFlt1, PI3K, Akt, and HIF-1 in placental samples from ten women with pre-eclampsia and ten normotensive control patients and in human choriocarcinoma trophoblast cells treated with 600 mu M CoCl2 by Western blotting. Using models of placental hypoxia, we also determined whether inhibition of the PI3K-Akt pathway plays a direct role in regulating the expression of sFlt1. Results: The VEGF, PIGF, sFlt1, PI3K, Akt, and HIF-1 levels were significantly higher in the preeclamptic placentas than the normal placentas. In the placental hypoxia models, the expression of VEGF and PIGF increased in a time-dependent manner, whereas the expression of sFlt1 plateaued after 3 h of CoCl2 treatment. The expression levels of p-Akt and PI3K were maximal after 6 and 12 h of CoCl2 treatment, respectively. The expression of HIF-1 cc increased in a time-dependent manner with CoCl2 treatment. Inhibition of the PI3K-Akt pathway with the PI3K-specific inhibitor LY294002 leads to decreased sFlt1 levels and unchanged or increased VEGF and PIGF levels. Conclusion: Inhibition of the PI3K-Akt pathway may be a useful therapeutic approach, if it were to decrease sFlt1 secretion without inhibiting VEGF or PIGF secretion. This pathway provides a potential target for a new treatment strategy in patients with pre-eclampsia. (C) 2010 Elsevier Ltd. All rights reserved.