Downregulation of cAMP-Dependent Protein Kinase Inhibitor-b Promotes Preeclampsia by Decreasing Phosphorylated Akt

Chunfeng Liu,Yiheng Liang,Shangrong Fan,Hao Wang,Siman Sun,Li Jiang,Mo Yang

doi:10.1007/s43032-020-00258-8

Abstract

Preeclampsia is a multi-system disease that is unique to human pregnancy. Impaired extravillous trophoblast migration and invasion accompanied by poor spiral vascular remodeling is thought to be the initial reason. This study investigated cAMP-dependent protein kinase inhibitor-b(PKIB) expression in placentas and its involvement in the pathogenesis of PE. We used immunohistochemistry and western blotting to calculate PKIB levels in the placentas. Then we knocked down PKIB by siRNA and used real-time cell analysis to assess the invasion and migration ability of trophoblasts. Tube formation assay and spheroid sprouting assay were utilized to identify the ability to form vessels of trophoblasts. At last, western blotting was used to demonstrate the level of phosphorylated Akt, as well as downstream-related genes of Akt signaling pathway in trophoblasts. We first found that PKIB expression level was lower in the PE placentas than in the normal placentas. In addition, we found that downregulation of PKIB can inhibit the migration, invasion, and the ability to form vessels of HTR8/SVneo cells. Downregulation of PKIB leaded to a decrease in phosphorylated Akt, as well as downstream proteins such as matrix metalloproteinase 2, matrix metalloproteinase 9, and glycogen synthase kinase 3β, which are related to migration and invasion. Our study revealed that the downregulation of PKIB expression resulted in decreased migration, invasion, and vessel formation ability by regulating Akt signaling pathway in placental trophoblasts in PE.

Highlights

Preeclampsia (PE) is a pregnancy-specific clinical syndrome, with hypertension and proteinuria after 20 weeks of gestation as the major clinical manifestation [1]
We first established a cell line in which protein kinase inhibitor-b (PKIB) was knocked down using siRNA in HTR8/SVneo, an extravillous trophoblasts (EVTs)-like first trimester trophoblast cell line
Spheroid sprouting assay confirmed that angiogenic properties of trophoblasts were decreased by knockdown of PKIB (Fig. 3d, e)

Summary

Introduction

Preeclampsia (PE) is a pregnancy-specific clinical syndrome, with hypertension and proteinuria after 20 weeks of gestation as the major clinical manifestation [1]. PE is thought to be a two-stage disorder: abnormal placental implantation in early pregnancy, followed by maternal inflammatory response [4, 5]. During placental implantation of a successful pregnancy, the EVTs penetrate the endometrium and the underlying myometrium to induce intimate interactions between the placenta and uterine wall [7]. In PE, the EVTs migration and invasion ability is impaired, followed by poor spiral vascular remodeling and decreased volume of maternal blood flowing into the uteroplacenta. This causes several negative consequences, including high pressure, abnormal stress in the villous placenta, increased shedding of necrotic placental debris into maternal vessels, and disruption of endothelium function [8,9,10]. The specific molecular mechanism of trophoblastic abnormalities leading to PE remains unclear

Methods

Results

Conclusion