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2017 ; 12
(3
): e0173711
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Alpha-1-antitrypsin suppresses oxidative stress in preeclampsia by inhibiting the
p38MAPK signaling pathway: An in vivo and in vitro study
#MMPMID28358847
Feng YL
; Yin YX
; Ding J
; Yuan H
; Yang L
; Xu JJ
; Hu LQ
PLoS One
2017[]; 12
(3
): e0173711
PMID28358847
show ga
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.