Alpha-1-antitrypsin suppresses oxidative stress in preeclampsia by inhibiting the p38MAPK signaling pathway: An in vivo and in vitro study.

Ya-Ling Feng,Ling-Qin Hu,Hua Yuan,Lan Yang,Yong-Xiang Yin,Yong-Xiang Yin,Jian Ding,Jian-Juan Xu,Ferenc Gallyas

doi:10.1371/journal.pone.0173711

Abstract

This present study was designed to investigate the effects of alpha-1-antitrypsin (AAT) on oxidative stress in preeclampsia (PE) by regulating p38 mitogen-activated protein kinase (p38MAPK) signaling pathway. HTR8/SVneo cells were randomly assigned into normal, hypoxia/reoxygenation (H/R), HR + AAT and HR + siRNA-AAT groups. Quantitative real-time polymerase chain reaction (qRT-PCR) and Western blotting were used to detect the mRNA and protein expressions of p-p38MAPK, AAT, signal transducer and activator of transcription 1 (STAT1) and activating transcription factor2 (ATF2). Flow cytometry, scratch test, cell counting kit-8 (CCK-8) assay and the 3-(4,5)-dimethylthiazol (-z-y1)-3,5-di- phenyltetrazolium bromide (MTT) assay were conducted to detect reactive oxygen species (ROS) and cell apoptosis, cell migration, proliferation and cytotoxicity, respectively. Mouse models in PE were established, which were divided into normal pregnancy (NP), PE and PE + AAT groups with blood pressure and urine protein measured. Chromatin immunoprecipitation (ChIP) and enzyme-linked immunosorbent assay (ELISA) were conducted to detect the activity of oxidative stress-related kinases and expressions of inflammatory cytokines and coagulation-related factors in cells and mice placenta. Immunohistochemistry, Western blotting and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay were performed to detect AAT and p38MAPK expressions, apoptosis-related protein expressions, and apoptosis rate in mice placenta. Compared with the normal group, the H/R group had decreased expression of AAT, activity of superoxide dismutase (SOD) and GSH-Px, cell proliferation and migration, but increased p38MAPK, STAT1, ATF2, MDA, H2O2, inflammatory cytokines, coagulation-related factors, cell cytotoxicity, ROS, apoptotic factors and apoptosis rate. Compared with the H/R group, the HR + ATT group had increased expressions of AAT, activity of SOD and GSH-Px, cell proliferation and migration but decreased p38MAPK, STAT1, ATF2, malonyldialdehyde (MDA), H2O2, inflammatory cytokines and coagulation-related factors, cell cytotoxicity, ROS, apoptotic factors and apoptosis rate, while opposite results were observed in the HR + siRNA-ATT group. Compared with the NP group, the PE group had decreased activity of SOD and GSH-Px but increased MDA, H2O2, AAT, p38MAPK, inflammatory cytokines, coagulation-related factors and apoptosis rate. The indexes in the PE + AAT group were between the NP and PE groups. Thus, we concluded that AAT suppressed oxidative stress in PE by inhibiting p38MAPK signaling pathway.

Highlights

Preeclampsia (PE) is a specific disorder related to pregnancy with a manifestation of proteinuria, gestational hypertension, and an excessive inflammatory response, as well as systemic endothelial cell activation [1]
The pro-oxidant (MDA) and antioxidant (GSH and superoxide dismutase (SOD)) balance is disturbed in pregnancy with PE, and increased oxidative stress along with antioxidative defense mechanisms may contribute to the progression of PE [5]
Our study aims to investigate the roles of AAT and the P38 mitogen-activated protein kinase (p38MAPK) signaling pathway in OS during PE

Summary

Introduction

Preeclampsia (PE) is a specific disorder related to pregnancy with a manifestation of proteinuria, gestational hypertension, and an excessive inflammatory response, as well as systemic endothelial cell activation [1]. As previous researches were mainly focused on the vascular endothelial cells and tissues or the relationship between AAT or p38MAPK signaling pathway and the development of PE, there was no study conducted regarding how AAT affects the development of PE through oxidative stress by regulating p38MAPK signaling pathway in trophoblast cells. This current study aims to explore the role of AAT and the p38MAPK signaling pathway in oxidative stress during the onset of PE and to investigate whether AAT treatment could be therapeutically beneficial to the improvement of PE

Methods

Results

Conclusion