SRI Supplement to Reproductive Sciences - Volume 25 Number 1 - March 2018 - 168A

168A

Reproductive Sciences Vol. 25, Supplement 1, March 2018

silver clips were placed on intra-abdominal fat. Rats were euthanised
on GD21, tissues collected and mesenteric artery function assessed by
pressure myography.
RESULTS: Litter size was reduced in both RUPP (n=5(3-10)) and sRUPP
(n=8(1-10)) compared to Sham (n=16(13-19)) while fetal body weight was
unaltered (Sham 5.17±0.32 g, RUPP 5.28±0.15 g, sRUPP 5.81±0.15 g).
Sham pregnant rats displayed reduced MAP in late gestation; approx. -4
mmHg (baseline: 98±1 mmHg vs. post-op: 94±1 mmHg). In both RUPP
and sRUPP animals, MAP rose by 12-16 mmHg (RUPP baseline: 90±1
mmHg vs. post-op: 102±2 mmHg and sRUPP baseline: 91±1 mmHg vs.
post-op: 107±1 mmHg). Correspondingly, mesenteric arteries from both
RUPP and sRUPP groups demonstrated reduced elasticity, distension
at 100 mmHg intraluminal pressure: Sham 76±3 μm, RUPP 64±4 μm,
sRUPP 65±15 μm.
CONCLUSION: In conclusion, the more specific sRUPP technique
shows promise as a refined model of PE which may avoid the confounding
issues of decreased hindlimb perfusion. This model further confirms the
specificity of the phenotypic maternal symptoms as being attributable to
reduced perfusion of the uteroplacental units, and not to a generalized
systemic ischemia, that therefore confirms this as a relevant and improved
model of PE.

T-176
L-Arginine during Pregnancy - A Preclinical Meta-Analysis on Fetal
Growth and Maternal Blood Pressure. Fieke Terstappen†,2 Nina D
Paauw,2 Wessel Ganzevoort,1 Jaap A Joles,2 Hendrik Gremmels,2 Anna T
Lely*.2 1Academic Medical Center, Amsterdam, Netherlands; 2University
Medical Centre Utrecht, Utrecht, Netherlands.
INTRODUCTION: L-Arginine supplementation during pregnancy has
been studied as a prenatal therapeutic and preventive approach for fetal
growth restriction (FGR) or preeclampsia (PE). While beneficial effects
on fetal growth and maternal blood pressure have been reported in human
studies, limitations in study design (small number of patients per study,
low risk group) hinders us to draw a clear conclusion. We conducted a
meta-analysis on the effect of L-arginine on fetal growth and maternal
blood pressure in preclinical and clinical studies. This combined animal/
human meta-analysis enables us to identify modifiable factors - such as
dose and administration strategy - to improve human RCT.
METHODS: Pubmed, Embase, and the Cochrane Library were search for
studies reporting on the effect of prenatal supplementation with L-Arginine
on fetal growth or maternal blood pressure. Ratio of mean birth weight and
mean differences in blood pressure were calculated. We conducted a metaanalysis with subgroup analysis (species, administration timing, route,
and scheme, prevention vs. treatment, model) to identify potential factors
that influence the efficacy of L-arginine the most. Dose-response curves
expressed in metabolic weight (mg/kg0.75/24h) were fitted using splines.
RESULTS: 30 animal (mouse, rat, pig, sheep, cow, horse) and 12 human
studies were included. This meta-analysis shows that L-arginine increases
fetal growth in FGR/PE compared to risk groups or healthy pregnancies
(1.13 [1.09;1.7] vs. 1.04 [0.99;1.10] vs.1.02 [0.99;1.04]; p=1e-9). The
effect was comparable between species, administration route and scheme,
but administration in middle or late pregnancy appeared to be more
effective than early or full gestation (p=9e-10). L-Arginine significantly
lowers maternal blood pressure in FGR/PE only (-22 [-32.4;-12.8] mmHg;
p=6e-6) with the decrease depending on the baseline blood pressure. The
effect was similar in administration timing and route, but stronger in rats
compared to humans and in continuous administration. There was no
dose-response effect in fetal growth or in blood pressure.
CONCLUSION: this meta-analysis supports that prenatal supplementation
with L-Arginine improves fetal growth and maternal blood pressure in
pregnancies complicated by FGR/PE. This combined meta-analysis shows
differences in blood pressure in continuous versus interval administration;
whether this is influenced by effect in species remains to be elucidated.
The use of L-arginine seems to be especially optimal for treatment rather
than prevention of placental insufficiency. Current doses used in studies
suffice to gain optimal effect for both fetal growth and blood pressure.

Scientific Abstracts

T-177
GNA14 Overexpression Alters FGF2-, but Not VEGFA-, Induced Fetal
Endothelial Function in Association with Elevated Phosphorylation
of PLCβ3. Qingyun Zou†, Chi Zhou†, Ai-Xia Liu*, Xin-Qi Zhong*, Qin
Yan*, Yan Li†, Jing Zheng*. University of Wisconsin-Madison, Madison,
WI, United States.
INTRODUCTION: During pregnancy, remarkable fetoplacental
vascular growth and development occur to support the growing fetus.
Interruption of such processes may cause adverse pregnancy outcomes i.e.
preeclampsia (PE) and IUGR. FGF2 and VEGFA are two key regulators
of fetoplacental endothelial function. G protein α subunit (Gα) mediates
many cellular signaling networks. G protein α subunit 14 (GNA14),
a member of Gαq/11 subfamily, is identified as a human hypertensionsusceptibility gene. Previously, we have reported that GNA14 is expressed
in villous endothelial cells and its protein levels are increased (~2.9 folds)
in placentas from PE vs. normal pregnancy. However, roles of GNA14 in
mediating FGF2- and VEGFA-induced fetal endothelial function remain
unclear. Here, we hypothesize that GNA14 overexpression impairs fetal
endothelial function.
METHODS: Primary human umbilical cord vein endothelial
cells (HUVECs) constantly cultured under a physiological low O2
condition (3% O2) were used as a fetal endothelial cell model. GNA14
overexpression was performed by transfecting adenoviral vectors carrying
GNA14 cDNA (Ad-GNA14). After transfections, FGF2- and VEGFAinduced cell migration and permeability were determined using transwell
system and Electric Cell-substrate Impedance Sensing (ECIS) system,
respectively. Protein levels of GNA14 and phosphorylation of PLCβ3,
ERK1/2, and AKT1 were analyzed by Western blotting.
RESULTS: Ad-GNA14, but not Ad-GFP (a control), at 5 MOI elevated
(p < .05) GNA14 protein levels by ~5.0 folds, comparable to the PEincreased GNA14 in placentas. In Ad-GFP, both FGF2 and VEGFA
stimulated (p < .05) cell migration. FGF2 strengthened (p < .05) cell
integrity, while VEGFA weakened (p < .05) it. Comparing with Ad-GFP,
Ad-GNA14 decreased (p < .05) FGF-induced cell migration by ~46%,
while increased FGF2-induced cell integrity by ~28%. In contrast, AdGNA14 did not affect any of these cell responses induced by VEGFA.
In addition, Ad-GNA14 elevated phosphorylation of PLCβ3 (Ser 1105;
an inhibitory site) by ~4.7 folds, but not PLCβ3 (Ser 537), in response
to FGF2. However, Ad-GNA14 did not alter phosphorylation of either
ERK1/2 or AKT1 in response to FGF2.
CONCLUSION: Elevated GNA14 protein levels impairs fetal endothelial
function, possibly leading to fetal endothelial dysfunction in PE. (NIH
HD38843 to JZ)

T-178
Preeclampsia Dysregulates Endothelial Function-Associated Genes/
Pathways in Fetal Endothelial Cells. Chi Zhou†,2 Qin Yan,2 QingYun Zou†,2 Chanel T Tyler,2 Xing-Qi Zhong,2 Ronald R Magness*,1
Jing Zheng*.2 1University of South Florida, Tampa, FL, United States;
2
University of Wisconsin-Madison, Madison, WI, United States.
INTRODUCTION: Preeclampsia (PE) is a leading cause of fetal and
maternal morbidity and mortality during human pregnancy worldwide.
Although PE is considered as a maternal endothelial disorder, emerging
evidence indicates that PE also adversely impacts fetal endothelial
function. Notably, PE-offspring are at increased risk of cardiovascular
disorders in adulthood, implicating that PE adversely programs fetal
vasculature in utero. To date, information on PE-dysregulated genes/
pathways in fetal endothelial cells is limited. We hypothesize that PE
dysregulates endothelial function-associated genes/pathways in fetal
endothelial cells, leading to fetal endothelial dysfunction.
METHODS: Highly purified, Passage 0 human umbilical vein endothelial
cells (P0-HUVECs) were isolated from normal term (NT, n=8; 39±0.5
wks) and PE (n=4; 37.5±0.5 wks) pregnancies immediately after
C-section. Transcriptomic profile differences between these NT and PE
P0-HUVECs were identified using RNAseq with Illumina TruSeq RNA
library prep kits and an Illumina HiSeq2000 sequencer. Bioinformatics
and statistical analysis were performed using skewer, STAR, RSEM, and
EdgeR software. Functional genomics analysis was performed to identify

Table of Contents for the Digital Edition of SRI Supplement to Reproductive Sciences - Volume 25 Number 1 - March 2018