e248 Abstracts
E-POSTERS
E P19 EXPERIMENTAL HYPERTENSION AND PHARMACOLOGY
P were higher. Higher mRNA and protein expression levels of NOX2 and NOX4,
O along with lower mRNA and protein expression levels of nephrin, were observed
S in the kidneys of WKY+HHcy and SHR+HHcy rats, relative to SHR and WKY
T ALTERED EXPRESSION OF LUNG EPITHELIAL ION AND FLUID
E TRANSPORTERS DURING PROGRESSION OF PULMONARY
R HYPERTENSION IN THE MONOCROTALINE MODEL
S Katarina Zirova, Zuzana Kmecova, Jana Veteskova, Martina Podzubanova, Jan
Klimas, Peter Krenek. Comenius University in Bratislava, Faculty of Pharmacy
- Department of Pharmacology and Toxicology, Bratislava, SLOVAK REPUBLIC
Objective: Recent studies have evaluated a significant incidence of pleural ef-
fusions in patients with idiopathic and heritable pulmonary arterial hypertension
(PAH) co-diagnosed with isolated right-sided heart failure (RHF). Coincidence of
pleural effusions and RHF in these patients significantly increased mortality rate
during follow-up. However, mechanism of pleural effusions formation has not
been fully explained. Therefore, we examined expression of lung epithelial ion
and fluid transporters responsible for alveolar fluid clearance (AFC), which may
contribute to lung fluid accumulation under pathological conditions and poten-
tially formation of pleural effusions.
Design and method: 12-weeks old male Wistar rats were subcutaneously in-
jected with monocrotaline (60 mg/kg, MCT) or vehicle (CON). The relative ex-
pression of AFC transporters: Na+/K+-ATPase alpha1, alpha2, beta1, beta3 sub-
units, epithelial sodium channel (ENaC) alpha, beta, gama subunits, cystic fibrosis
transmembrane conductance regulator (CFTR) were determined by RT-qPCR 1, 2,
4 weeks after MCT application and also in end-stage disease (animals sacrificed
after acute health deterioration, ~ 4 weeks after MCT application) where pleural
effusions were observed.
Results: Increase in lung weight was observed from second week after MCT
application and was more profound in end-stage disease (by 26 % compared to
fourth week, p<0.05). A downregulation of all three pore-forming subunits of
ENaC was observed in end-stage disease, accompanied by downregulation of
small beta3 subunit of Na+/K+-ATPase. Expression of catalytic alpha2 subunit
of Na+/K+-ATPase gradually decreased with disease progression from first week
after MCT (-80% in end-stage vs. CON, p<0.05). On the other hand, we observed
upregulation of CFTR (by 80%) and beta1 subunit of Na+/K+ATPase (by 79% vs.
CON, p<0.05) mainly in end-stage disease.
Conclusions: Downregulation of lung epithelial ion and fluid transporters con-
tributing to AFC is predominantly limited to higher stages of PH with a marked
downregulation in end-stage. These expression alterations may contribute to over-
all lung weight increase observed in end-stage disease. However, observed early
alpha2 subunit of Na+/K+-ATPase downregulation could participate on the patho-
genesis of PH while upregulation of beta1 subunit and CFTR in end-stage disease
could represent a compensatory mechanism.
WHAT IS THE MAJOR FACTOR ON THE KIDNEY INJURY INDUCED
BY HYPERTENSION OR HYPERHOMOCYSTEINEMIA
Rui Xu, Ning Gao. Hospital of Shandong First Medical University, Jinan, CHINA
Objective: As we all know kidney injury could be induced by hypertension
or hyperhomocysteinemia (HHcy). What is the major factor? HHcy plays a
synergistic role with hypertension in vascular injury; however, the relation-
ship between HHcy and hypertension in renal injury remains unclear. Here,
we sought to evaluate the relationship between HHcy and hypertension in the
context of renal injury and to elucidate the mechanism of action underlying
this relationship.
Design and method: Wistar Kyoto (WKY) and spontaneously hypertensive
(SHR) rats were randomized into WKY, WKY+HHcy, SHR, and SHR+HHcy
groups. Blood pressure, plasma homocysteine, creatinine, serum malondialde-
hyde (MDA), serum superoxide dismutase (SOD), and urinary albumin creatinine
ratio (UACR) were measured. mRNA and protein expression levels of NOX2,
NOX4, and nephrin in the kidneys were also examined.
Results: The WKY+HHcy group exhibited lower serum SOD levels, relative to
the WKY group, along with higher levels of both serum MDA and UACR. Simi-
lar effects were observed in the SHR+HHcy group, relative to the SHR group.
Copyright © 2021 Wolters Kluwer Health, Inc. All rights reserved
Copyright © 2021 Wolters Kluwer Health, Inc. All rights reserved.
Moreover, compared with the levels in the WKY+HHcy group, mean SOD lev-
els in the SHR+HHcy group were lower, while mean MDA and UACR levels
controls, respectively. Furthermore, the mRNA and protein levels of NOX2 and
NOX4 were higher and the mRNA and protein levels of nephrin were lower in the
SHR+HHcy group than in the WKY+HHcy group. In addition, compared with
WKY+HHcy group, the UACR level of SHR group was higher.
Conclusions: HHcy appears to synergistically increase hypertensive renal dam-
age by enhancing oxidative stress. However, hypertension remains a major risk
factor for renal impairment compared to HHcy.
RESVERATROL PROTECT AGAINST HOMOCYSTEINE-INDUCED
RENAL DAMAGE THROUGH ANTIOXIDANT ACTION IN
SPONTANEOUS HYPERTENSIVE RATS
Rui Xu1, Qingchen Hui1, Xuan Zhao2. 1Hospital of Shandong First Medical Uni-
versity, Jinan, CHINA, 2People’s hospital of Dongying, Dongying, CHINA
Objective: The aim of this study was to investigate the effect of resveratrol in
renal damage and the underlying mechanism induced by Hhcy in SHRs.
Design and method: Twenty-four SHRs were divided into control group, Hhcy
group, Hhcy + resveratrol (Hhcy+Res) group. Hhcy groups were given intraperi-
toneal injection of homocysteine. In addition, the Hhcy + Res group was given
resveratrol by gavage. The Systolic blood pressure (SBP), diastolic blood pres-
sure (DBP), plasma Hcy, serum malondialdehyde (MDA), superoxide dismutase
(SOD) and the urinary microalbumin (UACR) were measured. To quantify RNA
and protein expression levels, the mRNA and the protein expression of nephrin
and NADPH oxidase subunits NOX2 and NOX4 in the kidney were examined by
quantitative Real-time PCR and western blot. Renal histological or pathological
changes were observed under light and electron microscope.
Results: There was no difference in blood pressure before and after Hcy or Res
intervention among groups. The homocysteine levels in Hhcy and Res group were
significantly higher than that in control group, which showed that the model was
successfully built by intraperitoneal injection. The serum SOD level was signifi-
cantly decreased in Hhcy group, while it was increased in Res group. On the
contrary, the serum MDA and UACR levels of model group were significantly
increased. after the intervention of resveratrol, the MDA and UACR levels were
reduced. Furthermore, the expression levels of mRNA and the protein of NOX2
and NOX4 were elevated while those of the nephrin were reduced in Hhcy group.
Resveratrol significantly reduced the expressions of NOX2 and NOX4, and in-
creased the expression of nephrin in renal tissue of SHRs with Hhcy.
Conclusions: These results suggested that resveratrol may improve renal damage
in hypertensive rats with Hhcy by exerting oxidative stress beyond blood pressure.
CAFFEIC ACID AMELIORATES ANGIOTENSIN II-INDUCED
INCREASE IN BLOOD PRESSURE BY ACTIVATING VASCULAR
SARCO-/ENDOPLASMIC RETICULUM CA-ATPASE2A
Hao Wu1, Liting Zhang1, Peng Gao2, Daoyan Liu2, Zhiming Zhu2. 1Department
of Endocrinology and Nutrition, The 910th Hospital of PLA, Quanzhou, CHINA,
2Department of Hypertension and Endocrinology, Daping Hospital, Third Mili-
tary Medical University, Chongqing, CHINA
Objective: Many studies have focused on reporting the beneficial effect of con-
sumption coffee or caffeine on cardiovascular disease. However, caffeic acid, a
component of coffee, which has a completely different structure from caffeine,
was rarely studied in cardiovascular metabolic diseases. In this study, we investi-
gated the effect of caffeic acid on vascular function and blood pressure of mice.
Design and method: Tension of endothelium-free mesenteric arteries from
8-week-old male wild-type (WT) mice was measured using wire myograph.
Cytoplasmic Ca2+ and sarcoplasmic reticulum (SR) Ca2+ were measured from
primary vascular smooth muscle cells (VSMCs) using fura-2AM and mag-fura-
2AM, respectively. Molecular docking was performed by AutoDock to determine
the binding force and sites of caffeic acid with sarco-/endoplasmic reticulum
Ca-ATPase (SERCA, a channel transporting Ca2+ from the cytosol into the SR
 Abstracts e249

lumen. PDB: 2YFY). 8-week-old male SERCA2a knockout mice and WT litter-
mates were surgically implanted with mini-osmotic pumps filled with angiotensin
II (Ang II) solutions. Then these mice were fed with normal diet or 0.05% caffeic
acids in drinking water for 8 weeks. The tail-cuff blood pressure (BP) and 24-hour
ambulatory BP were measured.
Results: First, we found that caffeic acid relaxed mesenteric artery pre-contracted
with norepinephrine (NE) with half-effective dose of 26.7uM. Furthermore, pre-
treatment with 50uM caffeic acid for 5 min completely smoothed NE-induced
vasoconstriction. Specifically, we found that caffeic acid reduced intracellular
Ca2+ in a concentration-dependent manner as well as resisted the increase of
intracellular Ca2+ induced by thapsigargin, a SERCA inhibitor, whereas caffeic
acid increased SR Ca2+ uptake. Molecular docking revealed caffeic acid binds
to SERCA to form strong hydrogen bonds at ASN-359, TYR-389, ARG-604 and
ASP-738. Eventually, we found that compared with the normal diet group, the
intervention of caffeic acid significantly attenuated AngII-induced increases of
tail BP and ambulatory BP in mice, but the hypotensive effect of caffeic acid
disappeared in SERCA2a knockout mice.
Conclusions: These results suggest that caffeic acid has a beneficial effect on
vascular function and blood pressure by activating SERCA2a on VSMCs. Con-
sumption coffee or caffeic acid may be an effective lifestyle to prevent and treat
hypertension.
MICRORNA PROFILE IN HUMAN VASCULAR SMOOTH MUSCLE
CELLS FROM HYPERTENSIVE SUBJECTS: FOCUS ON OXIDATIVE
AND ENDOPLASMIC RETICULUM STRESS
Objective: Four different antihypertensive agent classes are equivalently recom-
mended in the guidelines for first-line treatment of arterial hypertension (AHT).
However, it is unclear, if one of these agents is more potent than the others to reach
blood pressure (BP) control. We sought to compare response rates and BP control
in these four classes.
Design and method: Patients with newly diagnosed grade 1 or 2 AHT on
24h-BP measurements (ABPM) were randomized in a 1:1:1:1 fashion to either
perindopril (ACE), olmesartan (ARB), amlodipin (CCB) or hydrochlorothiazide
(HCT). ABPM were checked at baseline (BL), after 4 weeks of half dose (TP1)
and after 4 weeks of full dose (TP2), if BP control was not reached after TP1.
Patients were classified as controlled if mean BP was < 130/80 mmHg, awake BP
< 135/85 mmHg and night BP < 120/70 mmHg.
Results: 80 patients were randomized: 20 (25.0%) to ACE, 20 (25.0%) to ARB, 21
(26.3%) to CCB, and 19 (23.8%) to HCT. Mean age was 48 (±14) years, mean BMI
26.5 (±3.7) kg/m2. Mean 24 h systolic BP (sBP) was 141.8 (± 9.1) mmHg, diastolic
BP (dBP) 87.8 (± 7.6) mmHg (Table). Mean decrease from BL to TP1 was -5.8 (±
10.9) and -2.7 (± 5.6) mmHg, from BL to TP2 -11.6 (± 11.2) and -5.6 (± 6.7) mmHg,
from TP1 to TP2 -5.8 (± 12.0) and -2.8 (± 5.4) mmHg, for mean 24 h sBP and dBP,
respectively. After TP1, 21 patients (26.3%) had a controlled BP overall (Figure). Af-
ter TP2, the number of controlled patients increased to 24 patients (30.0%, p=0.250,
Figure). Control rates were significantly lower for HCT in comparison to the other
agents for diastolic values after TP1 (p=0.013) and after TP1 and TP2 (p=0.003).

Yu Wang, Livia L. Camargo, Wendy Beatie, Martin Mcbride, Augusto C. Monte-

zano, Rhian M. Touyz. Institute of Cardiovascular and Medical Sciences, Univer-
sity of Glasgow, Glasgow, UNITED KINGDOM
Objective: miRNAs are important to phenotypic control and function of vascular
smooth muscle cells (VSMC). miRNA dysregulation is associated with vascular
dysfunction, which involves oxidative and endoplasmic reticulum (ER) stress. We
assessed VSMC miRNA profile in human hypertension focusing on oxidative and
ER stress pathways.
Design and method: VSMCs from small arteries from normotensive (NT) and
hypertensive (HT) subjects were used. miRNA profiling of 758 miRNAs was
performed using TaqMan advanced miRNA assay (TaqMan Low Density Array
Human microRNA). Expression of vascular genes and proteins was detected by
RT-PCR and immunoblotting respectively. Ingenuity Pathway Analysis (IPA) was
used for miRNA target prediction (high predicted results).
Results: miRNA array identified 25 miRNAs uniquely expressed in HT and 21
miRNAs uniquely expressed in NT (CT<30). Of the 332 miRNAs present in both
HT and NT, 60 miRNAs were significantly upregulated in HT (fold change >1.5),
136 miRNAs were significantly downregulated in HT (fold change >1.5). In HT,
mRNA levels of Nox1 (1.71 fold) and protein levels of Nox4 (1.59 fold) and Nox5
(2.04 fold) were increased, p<0.05. SOD2 (4.43 fold) and GPx1 (1.97 fold) pro-
tein expression were upregulated in HT, p<0.05. PERK (1.57 fold) and elF2a (2.31
fold) protein levels were increased in HT, while ERO1A expression was decreased
in HT (2.36 fold), p<0.05. IPA was performed for the genes differentially ex-
pressed in HT compared to NT. After inversely pairing the miRNAs to the changes
observed in gene and proteins expression, miR-505-5p (-2.13 fold), miR-324-5p
(-1.51 fold), miR-185-5p (-1.742) and miR-491-5p (-1.667), which are predicted
to target Nox5, were downregulated. miR-185-5p and miR-491-5p together with,
miR-125a-3p (-2.04 fold) and miR-634 (unique in NT, CT=27.36) were predicted
to target GPx1. ER stress was also targeted by downregulated miRNAs in HT,
such as miR-200b-3p (-28.57 fold), predicted to target elF2a.
Conclusions: We identify a cluster of downregulated miRNAs in human hyper-
tension that are linked to pathways that regulate oxidative and ER stress. These
findings provide new molecular insights into processes underlying VSMC redox-
sensitive dysfunction and may elucidate vascular miRNA signatures in human
hypertension.
Conclusions: Half dose of the most common antihypertensive treatments lead
BLOOD PRESSURE CONTROL-RATES THROUGH MONOTHERAPY to BP control in 26% of patients with grade 1 or 2 AHT. Control rates were not
WITH FOUR DIFFERENT ANTIHYPERTENSIVE AGENTS IN significantly improved when increasing to full dose. HCT was the least effective,
TREATMENT-NAÏVE PATIENTS WITH ARTERIAL HYPERTENSION whereas ACE, ARB and CCB were comparably effective.

Annina Vischer1, Thenral Socrates1, Michael Mayr1, Manuel Haschke2, Thilo

Burkard1,3. 1University Hospital Basel, Medical Outpatient Department, Basel,
SWITZERLAND, 2University Hospital Bern, Clinical Pharmacology and Toxicol-
ogy, Bern, SWITZERLAND, 3University Hospital Basel, Cardiology Department,
Basel, SWITZERLAND
DIFFERENT FORMS OF AFFERENT NERVE INPUT IN CARDIAC AND
RENAL DISEASE IN RATS?
Kristina Rodionova2, Tilmann Ditting2, Lisa Pickny1, Christian Ott2, Roland
Schmieder1, Mario Schiffer1, Kerstin Amann1, Roland Veelken2. 1University of
Erlangen, Erlangen, GERMANY, 2University of Erlangen, Paracelsus Private
Medical School, Erlangen, GERMANY

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