Fluid Shear Stress Promotes Placental Growth Factor Upregulation in Human Syncytiotrophoblast Through the cAMP-PKA Signaling Pathway

Edouard Lecarpentier; Anthony Atallah; Jean Guibourdenche; Marylise Hebert-Schuster; Sarah Vieillefosse; Audrey Chissey; Bassam Haddad; Guillaume Pidoux; Daniele Evain-Brion; Abdul Barakat; Thierry Fournier; Vassilis Tsatsaris

doi:10.1161/HYPERTENSIONAHA.116.07890

Fluid Shear Stress Promotes Placental Growth Factor Upregulation in Human Syncytiotrophoblast Through the cAMP-PKA Signaling Pathway

Hypertension. 2016 Dec;68(6):1438-1446. doi: 10.1161/HYPERTENSIONAHA.116.07890. Epub 2016 Oct 3.

Affiliations

¹ From the INSERM, UMR-S 1139, Paris, France (E.L., A.A., J.G., M.H.-S., S.V., A.C., B.H., G.P., T.F., V.T.); PRES Sorbonne Paris Cité, Université Paris Descartes, Paris, France (E.L., A.A., J.G., M.H.-S., S.V., A.C., T.F., V.T.); Port Royal Maternity, Department of Gynecology Obstetrics I, Centre Hospitalier Universitaire Cochin Broca Hôtel Dieu, Groupe Hospitalier Universitaire Ouest, Assistance Publique-Hôpital de Paris, France (E.L., V.T.); DHU Risques et grossesse, Paris, France (E.L., J.G., T.F., V.T.); PremUP Foundation, Paris, France (E.L., J.G., D.E.-B., T.F., V.T.); Laboratoire d'Hydrodynamique (LadHyX), CNRS, École Polytechnique, Palaiseau, France (A.B.); SDBA Centre Hospitalier Universitaire Cochin Broca Hôtel Dieu, Groupe Hospitalier Universitaire Ouest, Assistance Publique-Hôpital de Paris, France (M.H.-S.); Department of Obstetrics and Gynecology, Centre Hospitalier Intercommunal de Créteil, CRC CHI Creteil, University Paris Est Creteil, France (B.H.); Service d'hormonologie Centre Hospitalier Universitaire Cochin Broca Hôtel Dieu, Groupe Hospitalier Universitaire Ouest, Assistance Publique-Hôpital de Paris, France (J.G.); and INSERM, UMR-S 1180, Université Paris-Sud, Université Paris-Saclay, F-92296, Châtenay-Malabry, France (G.P.). edouard.lecarpentier@wanadoo.fr.
² From the INSERM, UMR-S 1139, Paris, France (E.L., A.A., J.G., M.H.-S., S.V., A.C., B.H., G.P., T.F., V.T.); PRES Sorbonne Paris Cité, Université Paris Descartes, Paris, France (E.L., A.A., J.G., M.H.-S., S.V., A.C., T.F., V.T.); Port Royal Maternity, Department of Gynecology Obstetrics I, Centre Hospitalier Universitaire Cochin Broca Hôtel Dieu, Groupe Hospitalier Universitaire Ouest, Assistance Publique-Hôpital de Paris, France (E.L., V.T.); DHU Risques et grossesse, Paris, France (E.L., J.G., T.F., V.T.); PremUP Foundation, Paris, France (E.L., J.G., D.E.-B., T.F., V.T.); Laboratoire d'Hydrodynamique (LadHyX), CNRS, École Polytechnique, Palaiseau, France (A.B.); SDBA Centre Hospitalier Universitaire Cochin Broca Hôtel Dieu, Groupe Hospitalier Universitaire Ouest, Assistance Publique-Hôpital de Paris, France (M.H.-S.); Department of Obstetrics and Gynecology, Centre Hospitalier Intercommunal de Créteil, CRC CHI Creteil, University Paris Est Creteil, France (B.H.); Service d'hormonologie Centre Hospitalier Universitaire Cochin Broca Hôtel Dieu, Groupe Hospitalier Universitaire Ouest, Assistance Publique-Hôpital de Paris, France (J.G.); and INSERM, UMR-S 1180, Université Paris-Sud, Université Paris-Saclay, F-92296, Châtenay-Malabry, France (G.P.).

PMID: 27698065
DOI: 10.1161/HYPERTENSIONAHA.116.07890

Abstract

The effects of fluid shear stress (FSS) on the human syncytiotrophoblast and its biological functions have never been studied. During pregnancy, the syncytiotrophoblast is the main source of placental growth factor (PlGF), a proangiogenic factor involved in the placental angiogenesis and the vascular adaptation to pregnancy. The role of FSS in regulating PlGF expression in syncytiotrophoblasts is unknown. We investigated the impact of FSS on the production and secretion of the PlGF by the human syncytiotrophoblasts in primary cell culture. Laminar and continuous FSS (1 dyn cm^-2) was applied to human syncytiotrophoblasts cultured in a parallel-plate flow chambers. Secreted levels of PlGF, sFlt-1 (soluble fms-like tyrosin kinase-1), and prostaglandin E2 were tested by immunologic assay. PlGF levels of mRNA and intracellular protein were examined by RT-PCR and Western blot, respectively. Intracellular cAMP levels were examined by time-resolved fluorescence resonance energy transfer cAMP accumulation assay. Production of cAMP and PlGF secretion was significantly increased in FSS conditions compared with static conditions. Western blot analysis of cell extracts exposed to FSS showed an increased phosphorylation of protein kinase A substrates and cAMP response element-binding protein on serine 133. FSS-induced phosphorylation of cAMP response element-binding protein and upregulation of PlGF were prevented by inhibition of protein kinase A with H89 (3 μmol/L). FSS also triggers intracellular calcium flux, which increases the synthesis and release of prostaglandin E2. The enhanced intracellular cAMP in FSS conditions was blocked by COX1/COX2 (cyclooxygenase) inhibitors, suggesting that the increase in prostaglandin E2 production could activate the cAMP/protein kinase A pathway in an autocrine/paracrine fashion. FSS activates the cAMP/protein kinase A pathway leading to upregulation of PlGF in human syncytiotrophoblast.

Keywords: calcium signaling; cell extracts; phosphorylation; placental growth factor; trophoblast.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Analysis of Variance
Cells, Cultured
Cyclic AMP Response Element-Binding Protein / metabolism*
Female
Humans
Immunoblotting
Immunohistochemistry
Membrane Proteins / genetics*
Placenta / metabolism
Placenta / physiopathology
Placenta Growth Factor / genetics*
Pregnancy
RNA, Messenger / metabolism
Real-Time Polymerase Chain Reaction / methods
Signal Transduction / genetics
Stress, Mechanical
Trophoblasts / metabolism
Up-Regulation

Substances

Cyclic AMP Response Element-Binding Protein
Membrane Proteins
PIGF protein, human
RNA, Messenger
Placenta Growth Factor