Abstracts e423
MODERATED POSTER SESSION 18
MPS 18-01 MODE OF ACTION OF RB150, AN
AMINOPEPTIDASE A INHIBITOR PRODRUG AS A
CENTRALLY-ACTING ANTIHYPERTENSIVE AGENT
IN DOCA-SALT HYPERTENSIVE RATS
Rda Hmazzou, Adrien Flahault, Yannick Marc, Catherine Llorens-Cortes.
Department of Physiology, Collge de France, France
Objective: In the brain, angiotensin III (AngIII) generated from angiotensin
II (AngII) by aminopeptidase A (APA) exerts a tonic stimulatory action on the
control of blood pressure (BP) in hypertensive rats. Inhibition of brain APA by
RB150, an orally active prodrug of the specific and selective APA inhibitor, blocks
brain AngIII formation and normalizes BP in hypertensive rats. Since following
blockade of brain AngIII formation by RB150, no AngII accumulation was ob-
served, we aimed to delineate if this treatment induces the activation of another
metabolic pathway of brain AngII, in particular angiotensin converting enzyme
type 2 (ACE2) which converts AngII into angiotensin 17 (Ang17).
Design and Method: For this purpose, RB150 and MLN4760, an ACE2 inhibi-
tor, were administered by intracerebroventricular (ICV) route after insertion of an
intracerebral cannula to alert DOCA-salt rats, a model of salt-dependent hyperten-
sion. Mean arterial blood pressure (MABP) was recorded via an arterial femoral
catheter. Maximal MABP decrease and area under the curve (AUC) of the varia-
tions in MABP were calculated.
Results: ICV administration of RB150 (100 g) significantly decreased MABP
compared to vehicle (23 3 mmHg vs 6 2 mmHg; AUC: 17.6 2 vs
2.1 3). Administration of MLN4760 (10 g) didnt induce any significant
change of MABP compared to vehicle (10 3 mmHg; AUC: 1.4 2). Co-
administration of MLN4760 (10 g) with RB150 (100 g) partially inhibits
RB150-induced BP decrease.
Conclusions: In conclusion, ICV administration of the ACE2 inhibitor alone has
no effect on BP indicating that in DOCA-salt rats, brain Ang17 does not exert a
tonic stimulatory effect on the control of BP. However, when conversion of brain
AngII into AngIII is blocked by RB150, there is an activation of the conversion
by ACE2 of brain AngII into Ang17, which by acting on the Mas receptor, may
participate to the RB150-induced BP decrease in hypertensive rats.
MPS 18-02 ENDOTHELIN-1 OVEREXPRESSION PRESERVES
ENDOTHELIAL FUNCTION IN MICE WITH
VASCULAR SMOOTH MUSCLE CELL-SPECIFIC
DELETION OF PPAR-GAMMA
Sofiane Ouerd1, Noureddine Idris-Khodja1, Michelle Trindade2, Jordan Gornitsky1,
Asia Rehman1, Stefan Offermanns3, Frank Gonzalez4, Tlili Barhoumi1, Pierre Para-
dis1, Ernesto Schiffrin1. 1Hypertension and Vascular Research Unit, Lady Davis
Institute for Medical Research, Canada, 2Department of Clinical Medicine, State
University of Rio de Janeiro, Brazil, 3Department of Pharmacology, Max-Planck-
Institute for Heart and Lung Research, Germany, 4Department of Metabolism, Na-
tional Cancer Institute, U.S.A.
Objective: Peroxisome proliferator-activated receptor (PPAR) agonists reduce
blood pressure (BP) and vascular injury in hypertensive rodents. Ppar inactiva-
tion in vascular smooth muscle cells (VSMC) using a tamoxifen inducible Cre-
Lox system enhanced angiotensin II-induced vascular damage. Transgenic mice
overexpressing endothelin (ET)-1 in the endothelium (eET-1) exhibit endothelial
dysfunction, increased oxidative stress and inflammation. We hypothesized that
inactivation of the Ppar gene in VSMC (smPpar/) will exaggerate ET-1-in-
duced vascular damage.
Design and Method: Eleven week-old male control, eET-1, smPpar/ and eET-
1/smPpar/ mice were treated with tamoxifen (1 mg/kg/day, s.c.) for 5 days and
sacrificed 4 weeks later. BP was measured by telemetry. Endothelial function and
vascular remodeling using pressurized myography, reactive oxygen species (ROS)
production by dihydroethidium staining, monocyte/macrophage infiltration by
immunofluorescence and mRNA expression by reverse transcription-quantita-
tive PCR were assessed in mesenteric arteries (MA) or perivascular fat (PVAT).
Spleen T cell and monocyte profiles were assessed by flow cytometry.
Results: Systolic BP was 20 mmHg higher in eET-1 and unaffected by Ppar
inactivation. MA vasorelaxation to acetylcholine was impaired 37% only in smP-
par/. Likewise, ET-1-induced contractions were enhanced only in smPpar/. small-angle X-ray scattering (SAXS) approaches, we investigated the effect of
Copyright 2016 Wolters Kluwer Health, Inc. All rights reserved
Copyright 2016 Wolters Kluwer Health, Inc. All rights reserved.
W
E
D
N
ROS levels were increased 1.7-fold in smPpar/ and 2.5-fold in eET-1/sm E
Ppar/. Monocyte/macrophage infiltration in PVAT was 2-fold higher in eET- S
1 and smPpar/, which was further increased 2-fold in eET-1/smPpar/. The D
percentage of CD11b+ cells was increased 2.3-fold in smPpar/ and further in-
creased 1.5-fold in eET-1/smPpar/. The percentage of Ly-6Chi monocytes was
A
increased ~1.8-fold in eET-1 and sm Ppar/ but not eET-1/smPpar/. The per-
Y
centage of T regulatory cells was increased 1.5-fold in smPpar/ and decreased
by 26% in eET-1, which was further decreased by 38% in eET-1/smPpar/. P
O
Conclusions: These results suggest that increased ET-1 paradoxically preserves
endothelial function in mice with inactivated VSMC Ppar despite enhanced
S
oxidative stress. Flow cytometry data indicate that infiltrating monocyte/macro-
T
phages in these mice might be anti-inflammatory. E
R
MPS 18-03 VITAMIN C PREVENTS DECREASE OF BLEEDING
TIME; AN EXPERIMENTAL STUDY USING MICE
INDUCED CIGARETTE SMOKE
Alles Firmansyah1, Zelly Dia Rofinda2, Erkadius Erkadius3. 1Department of
Clinical Student, Faculty of Medicine Andalas University, Indonesia, 2Depart-
ment of Clinical Pathology, Faculty of Medicine Andalas University, Indonesia,
3Department of Physiology, Faculty of Medicine Andalas University, Indonesia
Objective: Cigarette smoke is one of the leading causes of preventable deaths in
the world. World Health Organization (WHO) 2015 reported 6,000,000 people are
estimated to die annually from cigarette smoke, with over 600,000 deaths due to
exposure second-hand smoke. Cigarette smoke known to enhance Reactive Oxy-
gen Species (ROS). ROS could damage vascular and alter platelet reactivity that
makes decrease of bleeding time as an indicator of thrombosis risk factor effect.
Vitamin C as an essential antioxidant was proven to reduce oxidative stress due to
ROS. We aimed to prove that vitamin C could prevent decreases bleeding time of
mice induced cigarette smoke.
Design and Method: An experimental study with randomized pre test-post test
control group design was conducted on 21 mice divided equally into three groups.
Group I as control; group II was given cigarette smoke in 10 minutes per day, and
group III was given vitamin C 0,4 mg/gW/day + cigarette smoke in 10 minutes
per day, respectively, for two weeks. The bleeding time was determined by a tail
bleeding method on day 0 and day 15. T-paired and LSD post-hoc test were used
to analyze the data. A P-value less than 0.05 considered significant.
Results: The result showed that group I had not a significant difference in mean
of bleeding time from 60.16 3.27 to 57.61 4.88 seconds. Group II had a
significant difference in mean of bleeding time from 59.34 6.93 to 38.85
3.43 seconds. Group III had a significant difference in mean of bleeding time from
59.36 3.07 to 51.85 3.45 seconds. Group II and group III had a significant
difference in post-test mean of bleeding time (p = 0.000).
Conclusions: This research proves that vitamin C could prevent decreases of bleed-
ing time. Further study is needed find out the effective dose and its potency in human.
MPS 18-04 FASUDIL PREVENTS CORONARY
MICROVASCULAR ENDOTHELIAL DYSFUNCTION
AND CARDIAC CONTRACTILE DYSFUNCTION IN A
RAT MODEL OF HYPERTENSIVE DIABETES MORE
THAN LOSARTAN
James Pearson1, Hirotsugu Tsuchimochi1, Tadakatsu Inagaki2, Cheng-Kun Du1,
Dong-Yun Zhang1, Daryl Schwenke5, Amanda Edgley4, Keiji Umetani3, Naoto
Yagi3, Mikiyasu Shirai1. 1Department of Cardiac Physiology, National Cerebral
and Cardiovascular Center, Japan, 2Department of Vascular Physiology, National
Cerebral and Cardiovascular Center, Japan, 3Biomedical Imaging Center, Japan
Synchrotron Radiation Research Institute, SPring-8, Japan, 4St Vincent Hospital,
University of Melbourne, Australia, 5Department of Physiology, University of
Otago, New Zealand
Objective: Diabetes, hypertension and obesity are risk factors for the develop-
ment of heart failure. Rho kinase (ROCK) appears to drive the coronary and con-
tractile dysfunction in early diabetes and in hypertension. We hypothesised that
ROCK activation plays a critical role in diabetic heart failure.
Design and Method: Using synchrotron radiation microangiography and in vivo