Lecarpentier Edouard, Atallah Anthony, Guibourdenche Jean, Hebert-Schuster Marylise, Vieillefosse Sarah, Chissey Audrey, Haddad Bassam, Pidoux Guillaume, Evain-Brion Daniele, Barakat Abdul, Fournier Thierry, Tsatsaris Vassilis
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.).
Hypertension. 2016 Dec;68(6):1438-1446. doi: 10.1161/HYPERTENSIONAHA.116.07890. Epub 2016 Oct 3.
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) 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.
流体切应力（FSS）对人合体滋养层细胞及其生物学功能的影响尚未得到研究。在孕期，合体滋养层细胞是胎盘生长因子（PlGF）的主要来源，PlGF是一种促血管生成因子，参与胎盘血管生成及血管对妊娠的适应性变化。FSS在调节合体滋养层细胞中PlGF表达方面的作用尚不清楚。我们在原代细胞培养中研究了FSS对人合体滋养层细胞产生和分泌PlGF的影响。将层流和持续的FSS（1达因/平方厘米）施加于在平行板流动腔中培养的人合体滋养层细胞。通过免疫测定法检测PlGF、可溶性fms样酪氨酸激酶-1（sFlt-1）和前列腺素E2的分泌水平。分别通过逆转录聚合酶链反应（RT-PCR）和蛋白质印迹法检测PlGF的mRNA水平和细胞内蛋白水平。通过时间分辨荧光共振能量转移cAMP积累测定法检测细胞内cAMP水平。与静态条件相比，在FSS条件下cAMP的产生和PlGF的分泌显著增加。对暴露于FSS的细胞提取物进行蛋白质印迹分析显示，蛋白激酶A底物和丝氨酸133位点的cAMP反应元件结合蛋白的磷酸化增加。用H89（3 μmol/L）抑制蛋白激酶A可阻止FSS诱导的cAMP反应元件结合蛋白的磷酸化和PlGF的上调。FSS还触发细胞内钙流，这会增加前列腺素E2的合成和释放。COX1/COX2（环氧化酶）抑制剂可阻断FSS条件下细胞内cAMP的增加，这表明前列腺素E2产生的增加可能以自分泌/旁分泌方式激活cAMP/蛋白激酶A途径。FSS激活cAMP/蛋白激酶A途径，导致人合体滋养层细胞中PlGF上调。
