Hernandez Isabelle, Chissey Audrey, Guibourdenche Jean, Atasoy Roger, Coumoul Xavier, Fournier Thierry, Beaudeux Jean-Louis, Zerrad-Saadi Amal
Université de Paris, INSERM UMR-S 1139, 3PHM, F-75006 Paris, France.
UF d'hormonologie Adulte de Cochin AP-HP, Hôpitaux Universitaires, F-75006 Paris, France.
Antioxidants (Basel). 2021 Feb 12;10(2):281. doi: 10.3390/antiox10020281.
Preeclampsia, a hypertensive disorder occurring during pregnancy, is characterized by excessive oxidative stress and trophoblast dysfunction with dysregulation of soluble Fms-like tyrosine kinase 1 (sFlt-1) and placental growth factor (PlGF) production. Nicotinamide Adenine Dinucleotide Phosphate (NADPH) oxidase (Nox) is the major source of placental superoxide in early pregnancy and its activation with the subsequent formation of superoxide has been demonstrated for various agents including Transforming Growth Factor beta-1 (TGF-β1), a well-known p38 MAPK pathway activator. However, the bridge between Nox and sFlt-1 remains unknown. The purpose of this study was to explore the possible signaling pathway of TGF-β1/Nox/p38 induced sFlt-1 production in human chorionic villi (CV).
Human chorionic villi from first trimester placenta (7-9 Gestational Weeks (GW)) were treated with TGF-β1 or preincubated with p38 inhibitor, SB203580. For NADPH oxidase inhibition, CV were treated with diphenyleneiodonium (DPI). The protein levels of phospho-p38, p38, phospho-Mothers Against Decapentaplegic homolog 2 (SMAD2), and SMAD2 were detected by Western blot. The secretion of sFlt-1 and PlGF by chorionic villi were measured with Electrochemiluminescence Immunologic Assays, and NADPH oxidase activity was monitored by lucigenin method.
We demonstrate for the first time that NADPH oxidase is involved in sFlt-1 and PlGF secretion in first trimester chorionic villi. Indeed, the inhibition of Nox by DPI decreases sFlt-1, and increases PlGF secretions. We also demonstrate the involvement of p38 MAPK in sFlt-1 secretion and Nox activation as blocking the p38 MAPK phosphorylation decreases both sFlt-1 secretion and superoxide production. Nevertheless, TGF-β1-mediated p38 activation do not seem to be involved in regulation of the first trimester placental angiogenic balance and no crosstalk was found between SMAD2 and p38 MAPK pathways.
Thus, the placental NADPH oxidase play a major role in mediating the signal transduction cascade of sFlt-1 production. Furthermore, we highlight for the first time the involvement of p38 activation in first trimester placental Nox activity.
子痫前期是一种孕期发生的高血压疾病，其特征为氧化应激过度和滋养层细胞功能障碍，伴有可溶性Fms样酪氨酸激酶1（sFlt-1）和胎盘生长因子（PlGF）产生失调。烟酰胺腺嘌呤二核苷酸磷酸（NADPH）氧化酶（Nox）是孕早期胎盘超氧化物的主要来源，包括转化生长因子β1（TGF-β1，一种著名的p38丝裂原活化蛋白激酶（MAPK）途径激活剂）在内的多种因子已证实可激活Nox并随后形成超氧化物。然而，Nox与sFlt-1之间的联系尚不清楚。本研究的目的是探索TGF-β1/Nox/p38诱导人绒毛膜绒毛（CV）中sFlt-1产生的可能信号通路。
用TGF-β1处理孕早期胎盘（妊娠7 - 9周（GW））的人绒毛膜绒毛，或用p38抑制剂SB203580预孵育。为抑制NADPH氧化酶，用二苯碘鎓（DPI）处理CV。通过蛋白质免疫印迹法检测磷酸化p38、p38、磷酸化抗五聚体蛋白同源物2（SMAD2）和SMAD2的蛋白水平。用电化学发光免疫分析法测定绒毛膜绒毛分泌的sFlt-1和PlGF，并用光泽精法监测NADPH氧化酶活性。
我们首次证明NADPH氧化酶参与孕早期绒毛膜绒毛中sFlt-1和PlGF的分泌。事实上，DPI抑制Nox可降低sFlt-1并增加PlGF的分泌。我们还证明p38 MAPK参与sFlt-1分泌和Nox激活，因为阻断p38 MAPK磷酸化可降低sFlt-1分泌和超氧化物产生。然而，TGF-β1介导的p38激活似乎不参与孕早期胎盘血管生成平衡的调节，且未发现SMAD2与p38 MAPK途径之间存在相互作用。
因此，胎盘NADPH氧化酶在介导sFlt-1产生的信号转导级联反应中起主要作用。此外，我们首次强调p38激活参与孕早期胎盘Nox活性。
