bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 071: Cytokine-angiotensin II Interplay in Cyclophilin D-mediated Vascular Oxidative Stress and Hypertension
Authors: Nazarewicz Rafal , Dikalova Anna , Itani Hana , McMaster William , Bikineyeva Alfiya , Harrison David G , Dikalov Sergey
Published: July 2021

Vascular inflammation and oxidative stress interact in a feed-forward fashion to promote vascular disease and hypertension. We hypothesized that angiotensin II and inflammatory cytokines encountered in hypertension co-operatively induce superoxide (O
2

•

-
) production by mitochondrial complex I and that efforts to reduce complex I O
2

•

-
will reduce hypertension. Treatment of human aortic endothelial cells in culture with angiotensin II (10 nM), IL17A (10 nM) and TNFα (1 nM), factors known to contribute to the hypertensive milieu, co-operatively induced mitochondrial O
2

•

-
from 340 to 958 pmol/mg protein as measured by HPLC and MitoSOX. This response was abolished by the complex I inhibitor rotenone. We further tested a potential role of Cyclophilin D (CypD), the redox sensitive regulatory subunit of the mitochondrial transition pore in complex I O
2

•

-
production. Both the specific CypD inhibitor Sanglifehrin A and knockdown of CypD by siRNA prevented endothelial cell O
2

•

-
production in response to Ang/IL17/TNF. We also found that this cytokine-angiotensin II milieu induced S-glutathionylation of CypD and that scavenging mitochondrial H
2
O
2
with mitoEbselen prevents this and eliminates CypD dependent complex I O
2

•

-
production. We further studied the functional role of oxidative stress induced by Ang/IL17/TNF in isometric tension studies of mouse aortic rings. Twenty-four hour treatment of organoid cultured vessels with AngII/IL17/TNF reduced endothelium-dependent vasodilatation to acetylcholine and this was prevented by knockdown of CypD and was not observed in vessels of mice with overexpression of mitochondrial SOD or mitochondrial catalase. The in vivo role of CypD in regulation of vascular O
2

•

-
and blood pressure was further studied in mice infused with angiotensin II (490 ng/kg/min). Treatment with Sanglifehrin A after the onset of hypertension reduced blood pressure from 162 to 133 mmHg (P<0.01), reduced vascular O
2

•

-
and improved endothelium-dependent vasodilation. These studies have defined a novel role of Cyclophilin D as a cause of vascular dysfunction and hypertension and have provided a new target for treatment of this disease.
Abstract 092: Uterine Artery Dysfunction in ACE2 Deficient Mice is Associated with Placental Hypoxia and Reduced Umbilical Flow
Authors: Yamaleyeva Liliya M , Pulgar Victor M , Lindsey Sarah H , Varagic Jasmina , McGee Carolynne M , Yamane Larissa , Brosnihan K. Bridget
Published: July 2021

Intrauterine fetal growth restriction (IUGR), one of the serious complications of pregnancy, increases the risk for perinatal morbidity and mortality, and cardiovascular and renal abnormalities later in life. The components of the renin-angiotensin system (RAS) are important mediators of uteroplacental vasculature during pregnancy. The distribution of angiotensin converting enzyme 2 (ACE2), a key enzyme of the RAS, is described in the invading and intravascular trophoblasts, decidual cells and in the umbilical cord. Deletion of ACE2 restricts fetal weight gain and reduces fetal-to-placental weights ratio. ACE2 deficiency results in augmented uterine artery contractility to Ang II at mid-gestation. In this study we determined whether uterine artery dysfunction is associated with placental abnormalities in ACE2 KO mice at mid-gestation. Systolic blood pressures were higher in pregnant ACE2 KO vs. C57BL/6 wild type (WT) (102.3±5.1 vs. 85.1±1.9 mmHg). Urinary protein excretion, serum creatinine, mean kidney or heart weights were not different in ACE2 KO vs. WT. Fetal weight, normalized to tibia length, and pup-to-placental weight ratio were lower in ACE2 KO vs. WT (p<0.05). A higher sensitivity to Ang II (pD
2
8.64±0.04 vs. 8.5±0.03, p<0.05) concomitantly with a greater active uterine arterial tension tested as the response to KCl 75 mM (1.90±0.15 vs. 1.13±0.12 mN/mm, p<0.05) were associated with lower Ang II receptor 2 (AT2R) protein expression (0.2±0.01 vs. 0.4±0.01, p<0.05) in uterine arteries from pregnant ACE2 KO. In the placenta, trophoblast invasion as assessed by the distribution of cytokeratin in trophospongium and decidual areas was not different between ACE2 KO and WT mice. The staining for pimonidazole, a marker of placental hypoxia, was significantly increased in the trophospongium and placental labyrinth of the ACE2 KO vs. WT (p<0.05). Umbilical artery velocities measured using VEVO 2100 ultrasound were lower in the ACE2 KO vs. WT mice (82.2±13.1 vs. 162.6±35.4 mm/s, p<0.05). In summary, ACE2 deficiency induces uterine artery dysfunction associated with placental hypoxia and reduced umbilical flow. These changes develop before the major growth of fetus occurs and may contribute to the exacerbation of IUGR in the ACE2 KO mice.
Abstract 099: EGF Receptor Inhibitor Erlotinib Prevented Abdominal Aortic Aneurysm But Not Hypertension Induced By Angiotensin II Plus Beta-aminopropionitrile
Authors: Tsuji Toshiyuki , Obama Takashi , Fukuda Yamato , Elliott Katherine , Eguchi Satoru
Published: July 2021

Enhancement of the renin angiotensin II (AngII) system has been implicated in the development of abdominal aortic aneurysm (AAA). However, detailed molecular mechanism(s) by which AngII promotes AAA remain uncertain. We have demonstrated the critical role of a metalloprotease, ADAM17, in AngII-induced EGF receptor (EGFR) transactivation and subsequent hypertrophy in vascular smooth muscle cells (VSMC). In caveolin 1-/- mice, AAA formation induced by AngII plus beta-aminopropionitrile (BAPN), a lysyl oxidase inhibitor, was attenuated. The attenuation of AAA formation was associated with suppression of ADAM17 induction and EGFR activation. Others have reported that systemic ADAM17 silencing attenuated CaCl2-induced AAA formation in mice. We hypothesized that pharmacological inhibition of EGFR may prevent AAA but not hypertension in mice co-infused with Ang II and BAPN via suppression of ER/oxidative stress. To test this hypothesis, 8 week old mice were co-treated with AngII 1000 ng/kg/min (4 weeks) and BAPN 150 mg/kg/day (2weeks) with or without erlotinib (8 mg/kg/day), and AAA formation was evaluated by echo (internal diameter) and measurement (external diameter) of the aortae. In mice with the co-infusion, 64.3% (9/14) were dead due to aortic rupture/dissection. All surviving mice with co-infusion had AAA with max external/internal diameter (mm) of 2.12±0.31/2.06±0.47 vs 1.01±0.22/1.06±0.02 with saline infusion (p<0.01). In contrast, erlotinib-treated mice with co-infusion did not die or develop AAA. The max external/internal diameter (mm) of AA in this treatment was 1.19±0.18/1.06±0.10. In contrast, both erlotinib-treated and non-treated mice with the co-infusion developed hypertension assessed by telemetry (MAP mmHg: 147±7 vs 151±12). The EGFR inhibition was also associated with lack of EGFR activation, ADAM17 induction, ER/oxidative stress and extravascular fibrosis/matrix deposition. In conclusion, vascular EGFR activation appears to contribute to AAA formation but not hypertension induced by AngII plus BAPN. The mechanism by which EGFR promotes AAA seems to involve induction of ER stress and oxidative stress.
Abstract 224: Chronic Infusion Of Angiotensin-(1-7) Alleviates Cerebral Hemorrhagic Injury Compromised By Angiotensin II
Authors: Bihl Ji C , Xiao Xiang , Zhang Cheng , Chen Shuzhen , Wang Jinju , Zhao Bin , Chen Yanfang
Published: July 2021

Angiotensin-(1-7) [Ang-(1-7)] is a potential vascular protective peptide which counteracts the effects of Ang II. In this study, we investigated the role of Ang-(1-7) in hemorrhagic stroke. Adult male C57BL/6 mice implanted with telemetric probe for recording arterial blood pressure (BP) were divided into four minipump treatment groups: saline, Ang II, Ang II + Ang-(1-7), Ang II + Ang-(1-7) + A-779. After 2 weeks of treatment, mice were subjected to induction of hemorrhagic stroke by microinjection of

collagenase (type VII; 0.075 U in 0.5 μL) in striatum. Mice were sacrificed at twenty-four hours after ICH induction. Brains were dissected and cut into coronal slices for histological analysis of middle cerebral artery (MCA)remodeling and hemorrhagic size. The plasma levels of tumor necrosis factor-α (TNF-α), monocyte chemoattractant protein 1 (MCP-1) and interleukin (IL-8) were determined by ELISA and the levels of NFκB and inhibitor of kappa B (IκB) in cerebral microvasculature were determined by western blot. We found (Figure): 1) Ang II increased BP, MCA remodeling and hemorrhagic size. 2) Ang-(1-7) significantly decreased the effects of Ang II on MCA and hemorrhage without affecting the BP, which were abolished by A-779. 3) Ang-(1-7) induced a 26-32% reduction in plasma level of TNF-α, MCP-1 and IL-8 increased by Ang II. 4) Ang-(1-7) also counter-regulated Ang II-induced NFƙB up-regulation and IƙB down-regulation. In conclusion, Ang-(1-7) counteracts Ang II on vascular remodeling and hemorrhagic injury by alleviating NFκB-related inflammation.
Abstract 244: Increased Expression and Enhanced Vasorelaxation Activity of Specific Estrogen Receptor Subtypes in the Aorta and Mesenteric Vessels during Pregnancy
Authors: Mata Karina M , Li Wei , Reslan Ossama M , Siddiqui Waleed T , Opsasnick Lauren A , Khalil Raouf A
Published: July 2021

Pregnancy (Preg) is associated with hormonal and vascular changes, and estrogen (E2) may promote systemic vasodilation during Preg; however, the specific E2 receptor (ER), post-ER signaling mechanisms and vascular bed involved are unclear. To test if Preg is associated with distinct expression/activity of ERs in different blood vessels, BP and plasma E2 were measured in virgin and day-19 Preg rats, and the aorta, carotid, mesenteric and renal artery were isolated for measurement of ERα, ERβ and GPR30 expression, and the responses to E2 and specific ER agonists PPT (ERα), DPN (ERβ) and G1 (GPR30). BP was in Preg (89±6) < virgin (98±4mmHg), and plasma E2 was in Preg (120.5±5.8) > virgin (94.3±7.5pg/ml). Western blots revealed increased ERα and ERβ in aorta and mesenteric artery and GPR30 in aorta of Preg vs virgin. Immunohistochemistry revealed that the increases in ERs were mainly in intima and media. E2 and PPT caused greater relaxation of aorta of Preg (52.8±5.5, 49.3±11.4) than virgin (30.0±3.9, 19.3±3.8%) and of mesenteric artery of Preg (77.9±4.7, 75.4±4.5) than virgin (57.4±5.9, 46.5±9.5%), but similar relaxation in carotid and renal artery of Preg vs virgin. DPN and G1 caused greater relaxation in mesenteric and renal artery (15 to 30%) than aorta and carotid artery (<10%), but only aortic relaxation to G1 was in Preg (26.2±4.4) > virgin (5.3±6.7%). The NOS inhibitor L-NAME ± EDHF blocker tetraethylammonium or endothelium removal reduced PPT relaxation in aorta, suggesting an endothelium-dependent mechanism, but did not affect E2, PPT, DPN or G1-induced relaxation in other vessels, suggesting endothelium-independent mechanisms. PPT caused relaxation of Ca
2+
entry-dependent KCl contraction of mesenteric artery that was in Preg (69.7±5.5) > virgin rats (52.9±8.11%). Thus, during pregnancy, an increased ERα expression in endothelial and smooth muscle layers of aorta and mesenteric artery is associated with increased ERα-mediated relaxation via endothelium-derived vasodilators and direct inhibition of Ca
2+
entry pathways, supporting a role of aortic and mesenteric arterial ERα in pregnancy-associated systemic vasodilation. GPR30 may contribute to aortic dilation while the enhanced ERβ may mediate other genomic vascular effects during pregnancy.
Abstract 247: Gender-specific Angiogenesis Suppression By Btg2 May Be Mediated Through Suppressed Renin Expression
Authors: Stodola Timothy J , Didier Daniela , Flister Michael , Lazar Jozef , Greene Andrew
Published: July 2021

Angiogenesis is the formation of new microvessels from existing vascular beds. AngII, a downstream product of renin, has been shown to be essential mediator of skeletal muscle angiogenesis in Dahl Salt Sensitive (SS) and Sprague Dawley rats, C57BL/6 mice, and human endothelial cells
in vitro.
Using partial chromosome introgression from the Brown Norway (BN) rat into the SS rat we have created two congenic lines with small (<300 Kbp) BN substitutions that differ by 23 Kbp. The congenic regions define an angiogenesis locus that contains one gene,
Btg2
. Sanger sequencing revealed no sequence variants in exons of
Btg2
between the BN and SS strains. Angiogenesis was measured using an
in vivo
electrical stimulation model of one hindlimb, with the contralateral leg acting as the control, in males and females of both new congenic strains. Males from Btg2
BN
have angiogenesis (TA=20.0±4.5% increase in vessel density in stimulated leg relative to unstimulated leg, EDL=15.8±5.8%,) while Btg2
SS
males did not have angiogenesis (TA=5.1±2.2%, EDL=4.4±2.7%). Females of both strains had angiogenesis: Btg2
BN
(TA=15.8±4.3%, EDL=3.4%), Btg2
SS
(TA=14.5±2.0%, EDL=14.6±2.5%). Stimulation significantly increased
Btg2
expression in both males and females. Btg2
SS
males had a significantly greater increase in
Btg2
mRNA in the stimulated muscle relative to unstimulated than Btg2
BN
males (5.0±0.9 versus 2.1± 0.3), but there was no difference in stimulated females (Btg2
BN
2.5±0.2, Btg2
SS
3.2±1.5). To test the hypothesis that
Btg2
impacted renin expression, we cloned the renin proximal promoter into a vector to drive luciferase, and co-transfected HEK-293 cells with this and vector(s) expressing
Btg2
or
Hoxb9
and
Btg2
. Normalized luciferase activity was 4.52±0.30 (arbitrary units) with an empty vector control and suppressed with the
Btg2
vector to 1.45±0.07. Co-expression of
Btg2
and
Hoxb9
lead to a further reduction in luciferase activity to 0.40±0.04. These data suggest the elevated
Btg2
expression observed in Btg2
SS
strain may be acting to suppress renin expression through renin proximal promoter transcription factor
Hoxb9
, leading to an inhibited angiogenic response.
Abstract 250: 6β-hydroxytestosterone, A Cytochrome P450 1b1-derived Metabolite Of Testosterone, Mediates Increase In Thirst, Renal Dysfunction, And End Organ Damage Associated With Angiotensin II-induced Hypertension In Male Mice
Authors: Pingili Ajeeth K , Kara Mehmet , Jennings Brett L , Estes Anne M , Malik Kafait U
Published: July 2021

Recently, we showed that 6β-hydroxytestosterone (6β-OHT), a cytochrome P450 1B1 (CYP1B1)-derived metabolite of testosterone, contributes to the development of angiotensin II (Ang II)-induced hypertension and associated cardiovascular pathophysiology. In view of the critical role of Ang II in renal homeostasis and end organ damage, we determined the contribution of 6β-OHT to Ang II actions on water consumption and renal function in male

Cyp1b1
+/+

and

Cyp1b1
-/-

mice. Eight weeks old male

Cyp1b1
+/+

and

Cyp1b1
-/-

intact or castrated mice were injected with 6β-OHT (15 μg/g, i.p. every 3
rd
day) or vehicle (DMSO, 50 μl), and infused with Ang II (700 ng/kg/min) or vehicle for 2 weeks. Urine was collected for 24 hours on the final day of experiment. Castration attenuated Ang II-induced increase in water consumption and urine output, proteinuria and decrease in osmolality in both

Cyp1b1
+/+

, and

Cyp1b1
-/-

mice (Table 1). 6β-OHT did not alter Ang II-induced increase in water intake, urine output, proteinuria and decrease in osmolality in

Cyp1b1
+/+

mice, but restored these effects of Ang II in

Cyp1b1
-/-

or castrated mice (Table 1).
Cyp1b1
gene disruption or castration prevented Ang II-induced renal fibrosis, inflammation, and oxidative stress. 6β-OHT did not alter Ang II-induced renal fibrosis, inflammation or oxidative stress in

Cyp1b1
+/+

mice, however in

Cyp1b1
-/-

or castrated mice it restored these effects of Ang II. These data suggest that 6β-OHT, contributes to increased thirst, impairment of renal function and end organ damage associated with Ang II-induced hypertension in male mice, and that CYP1B1 could serve as a novel target for the treatment of renal disease and hypertension.
Abstract 254: Inflammasome Activity Is Essential For Deoxycorticosterone Acetate/salt-induced Hypertension In Mice
Authors: Krishnan Shalini M , Sobey Christopher G , Kemp-Harper Barbara , Chan Christopher T , Diep Henry , Dowling Jennifer , Pinar Anita , Mansell Ashley , Drummond Grant R
Published: July 2021

Inflammasomes are signalling complexes comprised of a NOD-like receptor protein (NLRP), an adapter protein (ASC) and caspase-1. Inflammasomes detect host-derived danger signals and induce inflammation via activation of caspase-1, which in turn cleaves the cytokines pro-interleukin(IL)-1β and pro-IL-18 into their active, pro-inflammatory forms. Hypertension is associated with chronic renal inflammation, but the role of the inflammasome in this process is not known. Hence, we tested whether deoxycorticosterone acetate (DOCA)/salt-induced hypertension in mice is associated with increased expression and/or activation of the inflammasome in the kidney, and assessed the impact of inhibition of inflammasome activity on blood pressure (BP) and markers of renal inflammation and fibrosis. Male C57BL6/J (wild type) and ASC
-/-
mice were uninephrectomised, implanted with a DOCA pellet (2.4 mg/d, 21 d,
s.c.
) and had their drinking water replaced with 1% saline (1K/DOCA/salt). Control mice had a kidney removed but received a placebo pellet and normal drinking water. 1K/DOCA/salt-treated mice had elevated systolic BP (146±4 mmHg) compared to control mice (115±2 mmHg; n=13-16; P<0.05). 1K/DOCA/salt-induced hypertension was associated with increased renal mRNA expression (fold-change vs control; n=7-9; P<0.05) of inflammasome subunits NLRP3 (2.3±0.2), ASC (2.8±0.6) and pro-caspase-1 (2.6±0.5), and the cytokine, pro-IL-1 (4.0±0.8). Moreover, protein levels of cleaved (active) caspase-1 and IL-1 were increased by 1.6±0.2- and 2.1±0.3-fold, respectively in kidneys of 1K/DOCA/salt vs control mice (n=6; P<0.05). ASC
-/-
mice, which lack an active inflammasome complex, displayed blunted hypertensive responses to 1K/DOCA/salt-treatment (140±3 mmHg) compared to wild types (155±8 mmHg; n=8-9; P<0.05). ASC
-/-
mice were also protected from 1K/DOCA/salt-induced increases in renal expression of the inflammatory genes IL-6, IL-17a, CCL2, ICAM-1 and VCAM-1, and accumulation of collagen. Thus, renal inflammation, fibrosis and elevated BP in response to 1K/DOCA/salt-treatment are critically dependent on inflammasome activity, highlighting this signalling complex and its cytokine products as potential therapeutic targets to treat hypertension.
Abstract 283: Angiotensin II Induces Oxidative Stress Through ADP-ribose-Transient Receptor Potential Melastatin 2-Dependent Mechanisms In Human Microvascular Endothelial Cells: Role Of AT2 Receptor.
Authors: Dulak-Lis Maria G , Nguyen Aurelie D , Jenkins Carol J , Montezano Augusto C , Touyz Rhian M
Published: July 2021

Endothelial dysfunction is associated with oxidative stress and dysregulation of calcium signalling. We previously demonstrated that calcium dependent Nox5 is regulated by Ang II and may be implicated in vascular damage in hypertension. Elevation of intracellular free calcium concentration ([Ca2+]i) influences oxidative stress, which may be related to activation of redox sensitive calcium channel TRPM2. Here, we postulated that in human microvascular endothelial cells (HMEC), Ang II induces ROS generation through TRPM2-dependent processes. HMECs were stimulated with either Ang II (100 nM) or H2O2 (1 mM). ROS production was measured by lucigenin (superoxide) and amplex red (H2O2); redox sensitive MAPKs/eNOS activation by immunoblotting; and AT1R/AT2R mRNA by Q PCR. In some experiments, the following inhibitors were used: aminoethoxydiphenyl borate (APB, TRPM2/SERCA/IP3R inhibitor), N (p amylcinnamoyl)anthranilic acid (ACA, TRPM2/PLA2 inhibitor), 8-bromo-cADP-ribose (8 Br-cADPR, ADPR antagonist), 3,4-Dihydro-5[4-(1-piperindinyl)butoxy]-1(2H)-isoquinoline (DPQ, poly-ADPR polymerase inhibitor) and PD123319 (AT2R antagonist). In hmECs, Ang II increased superoxide levels (1.9±0.2 fold, p<0.05 vs. vehicle), an effect blocked by APB, ACA, 8-Br-cADPR, DPQ and PD123319. In addition, Ang II-induced increase in H2O2 production (1.6±0.21 fold, p<0.05 vs. vehicle) was inhibited by AT2R antagonism. Ang II stimulation increased ERK1/2 activation (185%), but not p38, through AT2R-dependent mechanisms (p<0.05). AT2R, but not AT1R was expressed in HMECs. H2O2, an activator of TRPM2, decreased superoxide production (25%) and increased MAPKs (p38: 82%; ERK1/2: 40%; JNK: 36%), as well as, eNOS (24%) activation, p<0.05. In addition, ROS formation by Ang II was measured in the presence of L-type calcium channel blocker diltiazem, which also blocked Ang II effects on superoxide production. In conclusion, our data demonstrate that Ang II-induced oxidative stress and redox-sensitive MAPK activation in hmEC cells may involve TRPM2 and calcium-dependent signalling, which are mediated via AT2R. Our findings identify a novel signalling pathways whereby Ang II/AT2R regulates endothelial cell ROS generation through TRPM2.
Abstract 309: Sp-3 But not the NF-kB Transcription Factor Causes the Up-regulation of Angiotensin Type 1 Receptor Expression and Contributes to Hypertension in Aging FBN rats.
Authors: Saleem Mohammad , Asghar Mohammad
Published: July 2021

We recently reported that age-associated oxidative stress is causal to higher renal angiotensin Type 1 (AT1) receptor function and hypertension in aged Fisher 344 X Brown Norway (FBN) rats. We became interested in examining the mechanism of higher AT1 receptor function in the aging kidneys. Adult (3-month) and aging (21 month) FBN rats were subjected to conscious blood pressure measurement by telemetry approach. The levels of AT1 receptor mRNA in the kidney cortex was measured by qRT-PCR while nuclear Sp-3 and NF-kB-p65 redox-sensitive transcription factors were determined by western blotting. We found that blood pressure was higher in aged than in adult rats (adult vs. old: 110±1 vs. 130±1 mmHg) which was associated with higher AT1 receptor mRNA levels (adult vs. old: 1.51±0.72 vs. 7.86±1.03 DU), and nuclear levels of both Sp-3 (adult vs. old: 0.56±.01 vs. 1.54±.02 DU) and NF-kB-p65 (adult vs. old: 0.9±.01 vs. 1.5±0.01 DU). To further delineate whether sp-3 or NF-kB-p65 or both transcription factors are responsible for the up-regulation of AT1 receptor, human kidney (HK2) cells were transfected with Sp-3 and NF-kB-p65 plasmids. We found that Sp-3 plasmid but not NF-κB-p65 plasmid transfection caused an increase in the levels of AT1 receptor protein in HK2 cells (control vs. transfected: 135±22 vs. 235±10 DU). Furthermore, Sp-3 siRNA treatment resulted in the reduction of Sp-3 (control vs. transfected: 136±10 vs. 93±21 DU) and AT1 receptor protein levels (control vs. transfected: 270±38 vs. 172±201 DU) in HK2 cells. Our results suggest that sp-3 but not the NF-κB-p65 is involved in the up-regulation of renal AT1 receptor that may be contributing to hypertension in aging FBN rats.
Abstract 313: Interferon Regulatory Factor 1 Plays an Important Role in Inhibition of Vascular Remodeling Through Angiotensin II Type 2 Receptor
Authors: Nakaoka Hirotomo , Mogi Masaki , Suzuki Jun , Kan-no Harumi , Tsukuda Kana , Chisaka Toshiyuki , Kukida Masayoshi , Wang Xiao-Li , Bai Hui-Yu , Shan Bao-Shuai , Iwanami Jun , Horiuchi Masatsugu
Published: July 2021

Objective:
Transcriptional control of angiotensin II type 2 (AT
2
) receptor expression is not well known and we previously reported that interferon regulatory factor (IRF)-1 plays physiological roles in “growth”-regulated AT
2
receptor expression in fibroblast. We studied whether IRF-1 is involved in the attenuation of vascular remodeling in association with AT
2
receptor up-regulation.

Methods:
Inflammatory vascular injury was induced by polyethylene-cuff placement around the mouse femoral artery in male wild-type mice (WT: C57BL/6J strain), IRF-1 knockout mice (IRF-1KO) and AT
2
receptor-null mice (AT
2
KO) at 10 weeks of age. After cuff placement, each mice were treated with an intraperitoneal injection of compound 21 (C21), AT
2
receptor agonist, at the dose of 10 μg/kg/day or saline. Formalin-fixed, paraffin-embedded sections were prepared using femoral arteries 14 days after cuff placement and subjected to Elastica van Gieson staining for the evaluation of neointima formation. Superoxide anion production and mRNA expressions 7 days after cuff placement were evaluated by dihydroethidium staining and real-time quantitative RT-PCR respectively.

Results:
Neointima areas in the injured artery induced by cuff placement were significantly increased in vehicle-treated group of IRF-1KO and AT
2
KO compared with those in WT. Moreover, we observed that treatment with C21 attenuated neointima formation 76% in WT, but 45% in IRF-1KO. Oxidative stress was more enhanced in vehicle-treated group of IRF-1KO compared with WT. Treatment with C21 markedly inhibited oxidative stress in WT, but not in IRF-1KO. AT
2
receptor mRNA expression was significantly decreased in IRF-1KO compared with that in WT; however, IRF-1 mRNA expression did not differ between AT
2
KO and WT.

Conclusion:
These results indicate that IRF-1 up-regulates AT
2
receptor expression and thereby plays an important role in the inhibition of vascular remodeling, supporting the notion that IRF-1 is one of the key transcriptional factor for AT
2
receptor expression and that targeting the immune system is pivotal in the treatment of vascular diseases.
Abstract 319: Overexpression of microRNA-429 into the Renal Medulla Attenuated Salt-sensitive Hypertension in Dahl S Rats
Authors: ZHU QING , HU JUNPING , WANG ZHENGCHAO , LI PIN-LAN , LI NINGJUN
Published: July 2021

HIF prolyl-hydroxylase 2 (PHD2) is an enzyme to promote the degradation of transcription factor hypoxia inducible factor (HIF)-1α. We have previously shown that high salt intake stimulates the expression of microRNA (miR)-429, which promotes the decay of HIF prolyl-hydroxylase 2 (PHD2) mRNA, and that reduction of PHD2 mRNA level leads to the accumulation of HIF-1α and activation of many HIF-1α-regulated antihypertensive genes such as nitric oxide synthase (NOS) 2 and heme oxygenase 1 in the renal medulla. This miR-429-mediated regulation of PHD2/HIF-1α pathway is an important molecular adaptation to promote extra sodium excretion and maintain blood pressure. However the high salt-induced increase in the renal medullary miR-429 level was impaired in Dahl S rat, a salt-sensitive hypertension model. The present study determined whether overexpression of miR-429 would reduce the levels of PHD2 mRNA, increase the expression of HIF-1α target genes in the renal medulla, and consequently attenuate salt-sensitive hypertension in Dahl S rats. Renal medullary miR-429 levels were increased by 2-fold via transfection of miR-429-expressing plasmid into the renal medulla in Dahl S rats, which was accompanied by 40% (0.4 of 1) decrease in PHD2 mRNA levels and 2-fold increase in NOS2 mRNA expression compared with scramble-miR-treated rats. Functionally, chronic high salt-induced sodium retention was remarkably reduced from 28.6 ± 2.4 mmole/kg in control rats to 18.5 ± 1.6 mmole/kg in miR-429-treated rats. Furthermore, hypertension induced by 2-week high salt intake was significantly attenuated in miR-429-treated rats. The mean arterial pressure in these Dahl S rats was 111.8 ± 1.7 mmHg on a low salt diet, 143.7 ± 4.1 on a high salt diet, and 128.6 ± 2.2 on a high salt diet treated with miR-429 plasmids, respectively. These results suggest that the impaired miR-429-mediated PHD2 inhibition in response to high salt intake in the renal medulla may represent a novel mechanism for hypertension in Dahl S rats and that correction of this impairment in miR-429 could be a therapeutic approach for salt-sensitive hypertension.
Abstract 344: Endothelin-1 Overexpression Exaggerates Diabetes-induced Endothelial Dysfunction
Authors: IDRIS KHODJA Noureddine , Mian Muhammad Oneeb Rehman , Barhoumi Tlili , Ouerd Sofiane , Gornitsky Jordan , Paradis Pierre , Schiffrin Ernesto L
Published: July 2021

Objective:
Vascular disease associated with endothelial dysfunction is a major cause of morbidity in patients with type-1 diabetes. Endothelin (ET)-1 plays a role in diabetes-induced vascular complications, since ET-1 type A receptor blockade reduces diabetes-induced vascular injury. However, whether ET-1 contributes to diabetes-induced endothelial dysfunction remains unproven. We hypothesized that vascular ET-1 overexpression will exaggerate diabetes-induced endothelial dysfunction.

Methods:
Diabetes was induced by streptozotocin treatment (STZ, 55 mg/kg/day, ip) for 5 days in 6-week-old male wild-type (WT) mice and in mice overexpressing ET-1 restricted to the endothelium (eET-1). Mice were studied 14 weeks later. Blood was collected to determine glucose. Mesenteric artery reactivity and remodeling were evaluated using pressurized myography and aortic fibronectin expression by immnunofluorescence.

Results:
STZ-induced diabetes was confirmed by a 3-fold increase in glycemia in WT and eET-1 (
P
<0.001). Diabetes impaired endothelium-dependent relaxation (EDR) reponses to acetylcholine in WT (60.9±6.4% vs 83.9±3.4%,
P
<0.05) and eET-1 (48.6±5.1% vs 81.5±5.2%,
P
<0.001). EDR impairment was exaggerated in eET-1 compared to WT (
P
<0.05). Meclofenamic acid, an inhibitor of cyclooxygenase, increased EDR in eET-1 compared to WT (78.4±9.4% vs 66.7±3.2%,
P
<0.01), which was not observed in diabetic mice. L-NAME, an inhibitor of nitric oxide (NO) synthase, completely blocked EDR in WT, eET-1 and diabetic WT, but not in diabetic eET-1 (4.1±1.6%, 6.4±5.7%, 2.2±4.6% and 26.6±4.6%,
P
<0.05). Apamin plus Tram34, inhibitors of endothelium-dependent hyperpolarization inhibited EDR in the four groups. Endothelium-independent relaxation to sodium nitroprusside, a NO donor, was similar in the four groups. Diabetes reduced media/lumen in WT (2.7±0.3 vs 3.6±0.3,
P
<0.05) and eET-1 (2.9±0.2 vs 3.8±0.3,
P
<0.05). Diabetes decreased aortic fibronectin expression in WT (94.0±11.0 vs. 151.9±21.8 RFU/μm
2
,
P
<0.05) and eET-1 (66.3±8.7 vs. 146.6±20.7 RFU/μm
2
,
P
<0.05).

Conclusion:
ET-1 contributes to alterations in several pathways mediating endothelium-dependent relaxation in type-1 diabetes, leading to exaggerated endothelial dysfunction.
Abstract 414: Inhibition of Mitogen-Activated Protein Kinase (MAPK) Signaling Ameliorates Endoplasmic Reticulum (ER) Stress, Inflammation and Sympathetic Nerve Activity in Heart Failure Rats.
Authors: Wei Shun-Guang , Yu Yang , Zhang Zhi-hua , Weiss Robert M , Felder Robert B
Published: July 2021

Heart failure (HF) is characterized by increased sympathetic nerve activity, driven in part by increases in brain inflammation and renin-angiotensin system activity. Angiotensin II (ANG II) and pro-inflammatory cytokines (PICs) activate neuronal MAPK signaling, which has been implicated in sympathetic activation in HF. ANG II and PICs can also activate ER stress, which plays an important role in regulating sympathetic drive and cardiovascular function in ANG II hypertension. The present study was designed to determine whether ER stress occurs in cardiovascular regions of the brain in HF, and if so how MAPK signaling interacts with ER stress. HF and SHAM (n=6) rats were produced by coronary ligation or sham ligation, respectively. HF rats received a chronic (4 wk) intracerebroventricular (ICV) infusion (0.25 μl/hr) of vehicle (Veh, n=6) or an inhibitor (0.25μg/μl, n=6 in each group) selective for p44/42 MAPK (PD98059), JNK (SP600125), or p38 MAPK (SB203580). ER stress markers (GRP78, ATF-6, P58
IPK
and CHOP) and NF-κB components (NF-κB p65 and IκB-α) were measured in the subfornical organ (SFO) and the hypothalamic paraventricular nucleus (PVN) by real time PCR. Plasma norepinephrine (NE) was measured by ELISA. Compared with SHAM rats, Veh-treated HF rats had significantly (*p<0.05) higher mRNA levels for the ER stress markers in SFO (2-3 folds) and PVN (1.8-2.5 folds); these levels were lower (28-55%*) in the HF rats treated with the MAPK inhibitors, with SB203580 being most effective in SFO and PD98059 most effective in PVN. Veh-treated HF rats had higher (95 ± 11 %*) NF-κB p65 mRNA level and lower (46 ± 5 %*) IκB-α mRNA level in SFO and PVN, compared with SHAM rats; these indicators of increased inflammation did not occur in the HF rats treated with the MAPK inhibitors. The plasma NE level (ng/ml) was higher in Veh-treated HF (36.93 ± 2.60*) than SHAM (17.90 ± 2.59) rats; this increase was attenuated in the HF rats treated with PD98059 (13.64 ± 0.92*) and SB203580 (24.68 ± 2.71*). These data demonstrate that ER stress increases in the SFO and PVN of rats with ischemia-induced HF. Inhibition of brain MAPK signaling reduces brain ER stress and inflammation and decreases sympathetic excitation in HF. Brain MAPK signaling is a potential target for therapeutic intervention in HF.
Abstract 415: Metabolic And Cardiovascular Effects Of Cilia Ablation From The Leptin Receptor-containing Neurons
Authors: Guo Deng F , Morgan Donald A , Grobe Justin , Rahmouni Kamal
Published: July 2021

Genetic defects that cause ciliary dysfunction are associated with obesity and cardiovascular diseases in humans and animal models suggesting that functional cilia are important for metabolism and cardiovascular regulation. Cilia are membrane-bound, microtubular projections present on the surface of virtually all cell types including the neurons that control metabolic and cardiovascular functions such as those expressing the leptin receptor (LepRb). To examine the importance of cilia expressed in these neurons, we tested the effect of ablating cilia. For this, we generated mice lacking a key component of the intraflagellar transport (IFT) machinery, IFT88 protein, required for cilia formation, in the LepRb-containing neurons. Breeding ift88
flox
mice with LepRb
Cre
mice created mice deficient in ift88 gene only in the LepRb positive neurons as indicated by the pattern of tdTomato reporter expression. Importantly, ift88
flox
/LepRb
Cre
mice displayed an obesity phenotype as indicated by the increased (P<0.05) body weight (41±2 vs. 37±1. g in controls) and fat mass measured by MRI (11±2 vs. 5±1 g in controls) in 25 weeks old mice. We found that the obesity phenotype in ift88
flox
/LepRb
Cre
mice is not due to an increase in food intake (3.03±0.25 vs. 2.96±0.29 g in controls). In contrast, energy expenditure was reduced in the ift88
flox
/LepRb
Cre
mice as indicated by the decreased (P<0.05) O
2
consumption (1.71±0.1 vs. 2.01±0.08 mL/100g/min in controls) and heat production (5.07±0.4 vs. 5.98±0.27 kcal/kg/h in controls). Moreover, radiotelemetry measurement of core body temperature showed that ift88
flox
/LepRb
Cre
mice had lower 24 h body temperature (36.7±0.2

vs.

37.2±0.2
o
C in controls). These results indicate that low metabolic rate accounts for the development of obesity in ift88
flox
/LepRb
Cre
mice. Next, we assessed the hemodynamic consequence of ablating the ift88 gene in LepRb positive neurons. However, mean arterial pressure and heart rate were not significantly different in the ift88
flox
/LepRb
Cre
mice (106±7 mmHg and 635±17 bpm, respectively) compared to controls (110±4 mmHg and 679±15 bmp). These findings demonstrate that deletion of cilia in LepRb expressing neurons alter energy homeostasis, but not cardiovascular regulation.
Abstract 418: Substance P and Mast Cell Interaction Exacerbates Pressure Overload-Induced Cardiac Remodeling
Authors: Supowit Scott , Li Jianping , DiPette Donald
Published: July 2021

This study was designed to test the hypothesis that cardiac sensory nerves and mast cells play a critical adverse role in transverse aortic constriction (TAC) pressure overload-induced heart failure. Moreover, the mechanism is mediated by the activation of protease activated receptor 2 (PAR-2) receptors primarily located on cardiac sensory nerves and neurokinin 1 receptors (NK-1, substance P) located on mast cells and fibroblasts. Groups (n=6/group) of male PAR-2 receptor KO, NK-1 KO, and wild type (WT) C57BL6 mice (26-28 g) were subjected to either sham or TAC surgery on day 1. Echocardiographic, biometric, and immunohistological analyses were performed on all groups of mice on day 14. At this time point, none of the structural echocardiographic parameters were significantly different between the TAC WT and the TAC NK-1 KO and TAC PAR-2 KO mice, except for the systolic left ventricular internal diameter ( TAC WT, 3.0±0.2, TAC NK-1 KO, 1.5±0.2, and TAC PAR-2 KO, 1.8±0.3 mm (sham WT, 2.2±0.1)). In contrast, the fractional shortening (FS) was significantly greater in the TAC NK-1 KO mice (50±4%) and the TAC PAR 2 KO mice (46±6%) compared with the TAC WT mice (24±3%) (sham WT mice 40±2%). The ejection fraction (EF) was also significantly greater in the TAC NK-1 KO mice (82±4%) and PAR-2 KO mice (76±5%) compared to the TAC WT mice (48±4%) (sham WT mice 71±3%). In addition, the ratios of heart weight to body weight and lung weight to body weight were less in the TAC NK-1 KO and TAC PAR-2 KO mice compared to the TAC WT mice and the interstitial collagen fraction was significantly less in the TAC NK-1 KO mice (0.6±0.2%) and TAC PAR-2 KO mice (1.1±0.1%) compared to the TAC WT mice (1.6 ±0.4%). Therefore, these data indicate that cardiac sensory nerves (via PAR-2 receptors) and mast cells (via NK-1 receptors) play a critical adverse role in pressure overload-induced heart failure suggesting that antagonists of either or both receptors may be novel therapeutic agents for the treatment of pressure-induced heart failure.
Abstract 434: Exposure To Air Pollutants And Mortality In Hypertensive Patients According To Demography: A 10 Year Case-crossover Study
Authors: Wong Martin C , Tam Wilson W , Wang Harry H , Lao Xiang Qian , Kwan Mandy W , Zhang Daisy D , Chan Wai Man , Fan Carmen K , Cheung Clement S , Tong Ellen L , Cheung Ngai Tseung , Tse Shelly L , Yu Ignatius T
Published: July 2021

Background:
Relatively few studies have explored the relationship between air pollution and cause-specific mortality among hypertensive patients. This study evaluated whether short term exposures to nitrogen dioxide (NO2), ozone (O3), particulate matter <10 mm in diameter (PM10) were associated with higher risk of mortality among a large hypertensive patients.

Methods:
A total of 223,287 hypertensive patients who attended any public health-care services in Hong Kong and prescribed at least 1 antihypertensive agent for the first time between 2001 and 2005 were followed up until 2010. A time-stratified, bi-directional case-crossover design was adopted to study the association between environmental exposures and mortality outcomes.

Results:
For all-cause mortality, significant positive associations were observed for NO2 and PM10 at lag 0 to 3 days per 10 MIUg/m3 increase in concentration and the excess risks ranged from 1.187%- 2.501%. Significant positive associations were found for O3 at lag 1 and 2 days and the excess risks were 1.654% (95% C.I. 0.469%, 2.852%) and 1.207% (95% C.I. 0.025%, 2.404%), respectively. We found similarly positive associations between NO2 (excess risks: 1.786%-2.798%), PM10 (1.126%-1.847%), O3 (3.035%-3.355%) and mortality due to respiratory disease. In stratified analysis, these significant results were observed amongst those aged >65 years and in cold seasons only.

Conclusions:
Older hypertensive patients are susceptible to all-cause and respiratory disease-specific deaths from these air pollutants in cold weather. This implies urgent need for protective air pollution standards for this susceptible population.
Abstract 447: Ac-SDKP Suppresses TNFα-induced ICAM-1 Expression In Endothelial Cells Via Inhibition Of IκB Kinase And NF-κB Activation
Authors: Zhu Liping , Yang Xiao-Ping , Nakagawa Pablo , Rhaleb Nour-Eddine , Harding Pamela , Xu Jiang , Carretero Oscar A
Published: July 2021

N-acetyl-seryl-aspartyl-lysyl-proline (Ac-SDKP) is a naturally occurring tetrapeptide that prevents inflammation and fibrosis in hypertension and other cardiovascular diseases. We previously showed that Ac-SDKP decreased transcription factor NF-κB activation in angiotensin II-induced hypertension and also reduced intercellular adhesion molecule-1 (ICAM-1) expression in an experimental autoimmune myocarditis and hypertension animal model. However, the mechanisms by which Ac-SDKP down-regulated ICAM-1 expression are unclear. TNFα is a proinflammatory cytokine that induces ICAM-1 expression on different cell types. We hypothesized that Ac-SDKP suppresses TNFα-induced ICAM-1 via inhibition of IκB kinase (IKK) and subsequently by blockade of NF-κB activation. Human coronary artery endothelial cells were treated with Ac-SDKP or vehicle and then stimulated with TNFα (0.5 ng/ml). ICAM-1 protein expression and phosphorylation of IKK (p-IKK), inhibitory κB (p-IκB), p38 (p-p38) and ERK (p-ERK) were measured by Western Blot. Activation of NF-κB was determined by electrophoretic mobility shift assay (EMSA). ICAM-1 expression was virtually undetectable under basal conditions, but greatly increased by TNFα. Ac-SDKP dose-dependently suppressed TNFα-induced ICAM-1 expression (set at a value of 1.0 arbitrary units, AU) to 0.67±0.13 (p<0.05), 0.51±0.12 (p<0.01) and 0.39±0.09 AU (p<0.01) at 0.1 nM, 1 nM and 10 nM, respectively. In addition, Ac-SDKP (10 nM) inhibited TNFα-induced p-IKK from 1.0 to 0.71±0.02 AU (p<0.01). Ac-SDKP also inhibited TNFα-induced IKKβ expression from 1.0 to 0.64±0.12 AU (p<0.05), without affecting IKKα expression. Furthermore, Ac-SDKP inhibited TNFα-induced p-IκB from 1.0 to 0.54±0.03 AU (p<0.01). EMSA results showed that Ac-SDKP inhibited TNFα-mediated activation of NF-κB, which was 0.63±0.04-fold of TNFα-treated cells. However, Ac-SDKP had no effect on TNFα-induced p-p38 and p-ERK. Thus, we conclude that Ac-SDKP inhibits TNFα-induced IKK and subsequent degradation of IκB, thereby preventing NF-κB activation and ICAM-1 expression. These inhibitory effects are independent of p38 and ERK.
Abstract 469: Influences of Dietary Weight Reduction on the Atrial Natriuretic Peptide System in Obese Subjects
Authors: Haufe Sven , Engeli Stefan , Lichtinghagen Ralf , Utz Wolfgang , Schulz-Menger Jeanette , Luft Friedrich C , Birkenfeld Andreas L , Jordan Jens
Published: July 2021

In obese individuals, reduced atrial natriuretic peptide (ANP) bioavailability appears to promote arterial hypertension and type 2 diabetes likely through direct metabolic actions. Mid-regional pro-ANP is cleaved off during ANP processing, thus, serving as ANP release marker. We hypothesized that dietary weight reduction improves ANP release.

In 75 overweight and obese women and men who had lost >5% body weight during six months hypocaloric dieting (-9.1±3.8 kg), we analyzed venous mid-regional pro-ANP and natriuretic peptide receptor (NPR) expression in abdominal subcutaneous adipose tissue before and after the intervention. Participants underwent ambulatory blood pressure (ABPM) monitoring and cardiac magnetic resonance imaging.

ABPM decreased from 118±9/73±7 to 115±9/71±6 mm Hg and left ventricular mass from 92±20 to 86±19 g (p<0.05 for all). End-diastolic and end-systolic volumes remained unchanged. Mid-regional pro-ANP plasma concentrations did not respond to weight loss (before: 51±24 pmol/; after 53± 24 pmol/l) and increased similarly during exercise testing. Adipose NPR-A mRNA expression remained unchanged whereas NPR-C receptor mRNA decreased from 1.2±0.7 to 1.0±0.5 (p<0.05).

Our data suggest that dietary weight loss does not augment ANP release in overweight and obese subjects. However, weight loss could improve ANP bioavailability through decreased clearance. Possible mechanisms are reduced fat mass and reduced expression of adipose tissue ANP scavenger receptor NPR-C.
Abstract 470: Increasing Muscle Mass As A Therapeutic Target For The Cardiometabolic Syndrome
Authors: Stepp David , Qiu Shuiqing , Mintz James D , Fulton David
Published: July 2021

Increases in muscle mass, as observed with exercise, are positive influences of cardiometabolic health. Muscle mass is limited by myostatin and deletion of myostatin results in a hypermusclar phenotype that has been shown to improve glucose tolerance in obese mice. The effects on vascular and hepatic endpoints are unknown.

Hypothesis:
Increasing muscle mass by deletion of muscle growth negative regulator myostatin, improves NOX-limited vasodilation and hepatic steatosis. Myostatin deletion increased muscle mass in both lean (gastrocnemius 57.93%) and obese mice (gastrocnemius 79.64%). Fasting glucose, HbA1c and glucose tolerance are improved in obese myostatin null mice. Obese mice demonstrated increased markers of oxidant load and superoxide-mediated impairment of acetylcholine (Ach)-induced vasodilation compared to lean mice. Deletion of myostatin in obese mice improved Ach-induced vasodilation in mesenteric arteries without effects in lean mice. Response to nitroprusside were similar in all groups. Treatment with GKT(1х10-6 mol/L, NADPH oxidase inhibitor) restored impaired vasodilation in obese mice. Obese mice, with or without myostatin, displayed gross histological steatosis. Elevations in AST and ALT were evident in obesity and unaffected by myostatin deletion. In contrast, evidence of fibrosis (Gomori trichome, collagen mRNA expression) was reversed in obese, hypermuscular mice. Bilary dysfunction, as evidenced by increased in bilirubin, was improve in obese mice lacking myostatin as were plasma level of alkaline phosphatase. Taken together, these data suggest that increasing muscle mass by deletion of myostatin improves the metabolic syndrome as evidence by alleviation of glucose intolerance, endothelial dysfunction and hepatic fibrosis.
Abstract 474: Efficacy and Safety of LCZ696 Compared with Olmesartan in Japanese Patients with Systolic Hypertension
Authors: Rakugi Hiromi , Kario Kazuomi , Yamaguchi Masako , Okino Naoko , Gotou Hiromi , Zhang Jack
Published: July 2021

Objective:
To evaluate the efficacy and safety of LCZ696, a first-in-class angiotensin receptor neprilysin inhibitor (ARNI) vs olmesartan in Japanese patients with systolic hypertension (SH).

Methods:
Patients aged ≥20 yrs with mean sitting (ms) SBP ≥150-<180 mmHg were randomized to once-daily LCZ696 200 mg or 400 mg (forced-titrated from 1-week LCZ696 200 mg), or olmesartan 20 mg for 8 weeks. The primary objective of this study was to test if LCZ696 200 mg is superior over olmesartan in msSBP reductions from baseline at week 8. Secondary endpoints were reductions in msSBP with LCZ696 400 mg, reductions in msDBP and ms pulse pressure (PP), BP control rate, and safety for all treatment groups at week 8. Efficacy and safety was also assessed by age (<65 and ≥65 years).

Results:
A total of 1161 patients (LCZ696 200 mg, n=387; LCZ696 400 mg, n=385; olmesartan, n=389) were randomized. Demographic and baseline characteristics were generally comparable across the treatment groups with mean age of 58.7 yrs (<65 yrs, 771 [67.1%]; ≥65 yrs, 382[32.9%]). The msSBP reductions at week 8 with LCZ696 200 mg were statistically significantly superior to olmesartan and significantly greater with LCZ696 400 mg than olmesartan (Table). Reductions in msDBP and msPP were significantly greater and BP control rates were significantly higher with LCZ696 vs olmesartan. Greater BP and PP lowering with LCZ696 vs olmesartan was independent of age. The incidence of AEs was similar across the treatment and age groups.

Conclusion:
Treatment with LCZ696 is effective and generally well-tolerated in Japanese patients with SH regardless of age and LCZ696 200 mg showed superior BP lowering effect than olmesartan 20 mg.