bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 123: Difficult-To-Control Blood Pressure in Non-Diabetic Hypertensive Subjects is Associated With Upregulated Gene Expression of Inflammatory Pathways
Authors: Umanath Kausik , She Ruicong , Hassett Clare , Adrianato Indra , Yee Jerry , Levin Albert , Ortiz Pablo
Published: September 2019

Introduction:
Hypertension (HTN) causes cardiovascular disease. Despite this, many patients do not achieve blood pressure (BP) control. We hypothesized that non-diabetic hypertensive subjects with difficult-to-control BP will activate predictive signaling pathways.

Methods:
We collected demographics and 24-h and first-morning urine collections from 61 subjects enrolled in the Systolic Blood Pressure Intervention Trial (SPRINT). A validated instrument was administered to asses sugar sweetened beverage intake and converted to grams fructose per day. The BP-Difficult (BPD) group had SBP >140 mmHg or >120 mmHg for intensive group subjects,

or

BP at “goal” for the trial group and more than the median number of agents in each group in study as a whole. The “BP Easy” group (BPE) comprised the remainder of subjects. Mean BP from two visits before SPRINT termination established the study BP. Gene expression from transcriptomic sequencing of cellular RNAs of first-morning urine were assessed for differential expression (DE) genes (>1.5-fold change;
P
<0.01) by the Wilcoxon Rank-Sum test and pathway analysis. 354erd

Results:
Recoverable RNA from 51 subjects was present in 33 BPD and 18 BPE subjects. In the BPD group, 338 genes were upregulated and 3 downregulated; pathway analysis identified interferon, phosphoinositide 3-kinase, B cell receptor, and inducible nitric oxide synthase signaling as key upregulated pathways (
P
<0.05). Most upregulated genes were components of pro-inflammatory pathways: interferon (IFNAR1), interleukin-2 and -4 (IL2RG, IL4R), tumor necrosis factor receptor (TNFRSF1B), and infiltrating B lymphocytes (PIK3AP1). BPD subjects with low 24-h urine sodium (<200 mmol) and fructose intake (<40 g/day) had 59 DE genes: 57 upregulated and 2 downregulated. Significant pathways included STAT3 and Cdc42 signaling pathways (
P
<0.05), and IL2RG, MAPK14, TGFB1, and IFNAR1 were components of these pathways.

Conclusions:
Urinary cells from a difficult-to-control BP SPRINT cohort demonstrated molecular signatures associated with specific pro-inflammatory pathways and immune mediators.
Abstract 126: Role of
V
γ6
+
γδ T Cells in Angiotensin Il-Induced Hypertension and Vascular Injury
Authors: Mahmoud Ahmad , Caillon Antoine , Paradis Pierre , Schiffrin Ernesto L
Published: September 2019

Introduction:
Both innate and adaptive immune cells (such as T lymphocytes) have been shown to play a role in hypertension and vascular injury. Recently, we demonstrated that a small subpopulation of T cells considered "innate-like", expressing the γδ T-cell receptor (TCR) rather than the much more frequent αβ TCR, plays a key role in hypertension and vascular injury. The number and activation of γδ T cells were increased after 7 days of angiotensin (Ang) II infusion, and absence of γδ T cells prevented the development of hypertension, endothelial dysfunction of resistance arteries, and activation of CD4
+
and CD8
+
T cells in mice infused with Ang II for 14 days. γδ T cells can be subdivided according to the TCR variant (V) subtype that is generally specific for a tissue. A subpopulation of γδ T cells in the lungs and skin that are Vγ6
+
and produce interleukin (IL)-17A was shown to respond promptly to pneumococcal infection and skin inflammation. However, γδ T cell Vγ subtypes involved in hypertension are still unknown. We hypothesized that γδ T cells involved in hypertension are Vγ6
+
.

Methods:
Eleven- to 13-week old C57BL/6J male mice were infused or not with Ang II (490 ng /kg /min, sc) for 14 days (n = 5-14), and γδ T cell Vγ subtypes were profiled by flow cytometry in the spleen, mesenteric lymph nodes (MLNs), thoracic aortic (TA), perivascular adipose tissue (PVAT) and mesenteric artery (MA) PVAT.

Results:
In spleen and MLNs the most abundant γδ T cell Vγ subtypes were Vγ1,2
+
and Vγ4
+
followed by Vγ6
+
, Vγ5
+
and Vγ7
+
. In TA PVAT, the most abundant γδ T cell Vγ subtype was Vγ6
+
followed by Vγ4
+
, Vγ1,2
+
, Vγ5
+
and Vγ7
+
. In MA PVAT, the most abundant γδ T cell Vγ subtype was Vγ6
+
followed by Vγ4
+
, Vγ7
+
, Vγ5
+
and Vγ1,2
+
. Ang II infusion increased the frequency of Vγ6
+
γδ T cells in the spleen (% of CD3
+
T cells: 0.77±0.09 vs. 0.5±0.02,
P
<0.01) and TA PVAT 5.9±0.8 vs. 3.6±0.4,
P
<0.01), whereas it only tended to increase in MA PVAT 14.5±1.7 vs. 10.2± 1.9,
P
=0.07). The Vγ6
+
γδ T cell frequency in MLNs was unaffected by Ang II infusion.

Conclusion:
A differential distribution of γδ T cell Vγ subtypes was observed in lymphoid organs compared to PVAT. Vγ6
+
γδ T cells may play a role in hypertension. Targeting Vγ6
+
γδ T cells could be a therapeutic approach to reduce inflammation in hypertension.
Abstract 131: Insulin Regulated Aminopeptidase Inhibitors are More Effective Than the ACE Inhibitor, Perindopril in Preventing Unilateral Ureteral Obstruction-Induced Renal Fibrosis in Mice
Authors: Chelvaretnam Sharenya , Shen Matthew , Mountford Simon J , Thompson Philip E , Chai Siew Yeen , Widdop Robert E , Samuel Chrishan S , Gaspari Tracey A
Published: September 2019

New strategies are needed to combat pathological organ fibrosis that leads to end stage organ failure. We have identified the enzyme insulin regulated aminopeptidase (IRAP) as one such promising new target, with IRAP KO mice completely protected, and pharmacological inhibition of IRAP reversing, age-induced cardiac fibrosis. However, the effect of IRAP inhibition in renal fibrosis is unknown. Therefore the objective of this study was to: (1) compare anti-fibrotic efficacy of 2 chemically distinct IRAP inhibitors, HFI-419 and SJM4-164 to the ACE inhibitor, perindopril (PER) in a murine model of unilateral ureteral obstructive (UUO)-induced renal fibrosis; and (2) investigate role of IRAP substrates in mediating ant-fibrotic effect of IRAP inhibitors. Male C57Bl/6J mice (8 weeks old) were assigned to one of the following 7 day protocols (n=6/gp): sham, UUO+Veh, UUO+HFI (0.72mg/kg/d), UUO+SJM (0.72mg/kg/d), or UUO+PER (1mg/kg/d). IRAP substrate involvement was determined using specific receptor blockers for the oxytocin receptor (OTR; Atosiban; 0.6mg/kg/d) or the AT2 receptor (AT2R; PD123319; 10mg/kg/d). Interstitial fibrosis was significantly increased in UUO+Veh-injured kidneys compared to sham (% positive staining: UUO=5.9±0.7 vs Sham=1.2±0.3; p<0.05). HFI and SJM treatment were equally effective in preventing UUO-induced increase in interstitial renal fibrosis (% positive staining: HFI=2.5±0.3, SJM=2.8±0.3, all p<0.05 vs UUO). PER had no effect on UUO-induced increase in interstitial renal fibrosis (PER=6.6±0.7), however it was as effective as the IRAP inhibitors in reducing markers of inflammation, including NFκB activation and macrophage infiltration. The anti-fibrotic effects produced with IRAP inhibition were prevented by OTR or AT2R blockade (% positive staining: OTR=5.0±0.55, AT2R=6±0.8). Overall, this study demonstrated greater reno-protection with IRAP inhibition compared to that achieved using an ACE inhibitor, with IRAP inhibitors exhibiting potent anti-fibrotic and anti-inflammatory effects. Moreover, for the first time we show that the anti-fibrotic effects of IRAP inhibition involve activation of receptors associated with endogenous IRAP substrates.
Abstract 142: Purinergic P2 X Receptors and a T 1 Receptors Share Post - Receptor Signaling Pathways Regulating Renal Afferent Arterioles in Angiotensin I I ( Ang I I) Dependent Hypertension
Authors: Kulthinee Supaporn , Shao Weijian , Franco Martha , Navar L.Gabriel
Published: September 2019

In Ang II dependent hypertension, the increased afferent arteriolar resistance (AAR) is assumed to be primarily due to the increased intrarenal levels of Ang II activating AT1 receptors. Because the renal interstitial ATP levels increase with increased renal perfusion pressure (RPP), purinergic P2X receptors (P2XR) also influence AAR. However, the interactions between the respective receptors has not been fully established. Accordingly, experiments were performed to test the hypothesis that P2XR and AT1R share post-receptor signaling pathways which allow dominance of P2XR to control AAR. Experiments using the in vitro juxtamedullary nephron preparation allowed direct visualization of afferent arteriolar diameters (AAD). Normotensive and Ang II infused hypertensive rats showed AAD responses to increases in RPP from 100 to 140 mmHg by decreasing AAD by 26% ± 10% and 19% ± 4% demonstrating autoregulation. In kidneys from normotensive rats, superfusion at 100 mmHg RPP with 5 μM ATP induced a rapid vasoconstriction from a control of 14.5 ± 0.9 μm to 11.2 ± 0.7 μm (n=8, P < 0.05). P2X1 receptor inhibition with NF449 (1 μM) increased AAD by 28% ± 7% returning it close to control values (14.0 ± 0.8 μm). P2X7 receptor inhibition with A438079 (1 μM) did not significantly alter the AAD (12.2 ± 0.6 μm), but combined treatment with P2X1R plus P2X7R inhibitors fully returned the AAD to control values (14.8 ± 0.7 μm), reflecting a dominant role of P2X1R in mediating ATP afferent vasoconstriction in normotensive kidneys. In Ang II infused hypertensive rats, at RPP 140 mmHg, treatment with AT1R inhibitor by SML1394 (1 μM) increased AAD by 10% ± 7% (13.44 ± 0.73 μm vs. 12.21 ± 0.63 μm, n=10, P<0.05). Treatment with P2X1R inhibitor increased AAD by 21% ± 14% (14.42 ± 0.51 μm vs. 11.94 ± 1.20 μm, n=5, P<0.05), and treatment with the P2X7R inhibitor also dilated AAD by 15% ± 3% (12.98 ± 0.64 μm vs. 11.26 ± 0.61 μm, n=5, P<0.05) reflecting a greater role of P2X7R in hypertension. Importantly, treatment with P2X1R inhibitor plus P2X7R inhibitor fully restored AAD to control levels (15.00 ± 0.65 μm vs. 12.02 ± 0.70 μm, n=5, P<0.05). The results suggest that in Ang II dependent hypertension, P2X1R, P2X7R and AT1R act through post-receptor converging signaling pathways allowing dominance of P2XR to regulate AAR.
Abstract 146: Glomerular Capillary Hypertrophy is Insufficient to Mitigate Mechanical Stresses in 5/6 Nephrectomy: A Modeling Study
Authors: Richfield Owen , Navar Luis G , Cortez Ricardo
Published: September 2019

The 5/6-nephrectomy (5/6 Nx) model of renal mass reduction shows an increased glomerular pressure (ΔP) and afferent plasma flow (Q
a
) in the remaining glomeruli. These hemodynamic consequences are assumed to increase glomerular capillary wall circumferential hoop stress (HS) and shear stress (SS), respectively. Due to a lack of anatomically-accurate mathematical models of the glomerulus, the actual magnitudes of SS and HS in the entire network of glomerular capillaries remain uncertain in 5/6-Nx. These forces are of significance due to the sensitivity of endothelial cells to SS and podocytes to HS.

We developed a mathematical model of blood flow through an anatomically-accurate rat glomerular capillary network to estimate the magnitudes of SS and HS on the glomerular capillary walls in 5/6-Nx. Filtration rates, wall HS and wall SS were calculated for each capillary segment of the network based on parameters obtained from a micropuncture study using normal rats and rats with 5/6-Nx (Kasiske, Bertram L., et al.
Circ. Res.
62.2: 367-374. (1988)). In this study, 5/6-Nx increased Q
a
and ΔP by 174.8% and 19.0%, respectively. To simulate glomerular capillary hypertrophy in 5/6-Nx, the capillary diameters were all increased by 16.1%. Using these data, the Young-Laplace equation was used to calculate HS in each capillary, and SS was calculated assuming Poiseuille flow with blood viscosity scaled based on vessel diameter and hematocrit.

Taking into account an increase in diameter of the glomerular capillaries, the model predicted mean SS values of 48.1 dynes/cm
2
in control and 79.4 dynes/cm
2
in 5/6-Nx. The increase in diameter of the glomerular capillaries was not sufficient to prevent the increase in SS, a finding in contrast to the previous experimental analysis which suggested normalization of SS. The increase in glomerular capillary diameter increased mean HS from 144.1 kPa to 199.2 kPa. This study demonstrates that this increase in diameter was not sufficient to mitigate the increased SS in 5/6-Nx and contributes to a significantly increased HS. These increased mechanical stresses on the glomerular capillaries may play a causative role in the progression of glomerular injury in 5/6-Nx thus leading to chronic kidney failure.
Abstract P102: A Vegan Approach for Gestational Hypertension in African Americans
Authors: Moreno Laura
Published: September 2019

The mortality rates in the African American infant population are higher than average. Non-Hispanic black infants died 11.4 to 1000 live births in 2016. The majority of complications stem from uncontrolled gestational hypertension. The purpose of this paper is to introduce the vegan diet with supplemental prenatal vitamins as a solution to a national crisis of infant mortality in the African American communities that would decrease gestational hypertension and its complications thereby improving maternal and fetal outcomes.The purpose of this paper is to introduce a research review that would strengthen the Vegan concept to prevent gestational hypertension and its complications in African Americans thus improving maternal and fetal outcomes.

Perhaps the ancestors of African Americans, who originally lived in hot, arid climates wherein sodium conservation was important for survival, evolved the physiologic machinery which protected them in their original habitat but makes it difficult for them to handle the excessive sodium they ingest when they migrate. African Americans tend to have higher blood volume, lower plasma renin activity as compared to white population (Langford, 1981). If gestational hypertension is genetic to all Africa, we would expect to see poor outcomes for black women

everywhere, but studies have found that foreign-born black women living in the United States have birth outcomes almost identical to white American women’s. Only descendants of the slave trade carry the hypertension gene and it has been shown to also exist in Western African countries, where Americans collected slaves. In the United States the Western diet consisting of a high content of sugars, fats and oils have pulled many triggers in our body’s responses, one of them being hypertension. Dietary fat reduction is critical in reducing insulin resistance which leads to hypertension. By educating the African American population on how a vegan diet can decrease both the maternal and neonatal mortalities, a significant improvement in mortality can be achieved.
Abstract P104: Activation of Endoplasmic Reticulum Stress and Thrombospondin-1 Anti-Angiogenic Mechanisms in Placentas of Hypertensive Mice
Authors: Marc Casandra , Achille Allahnah , Raguema Nozha , Pepin Émilie , Lavoie Julie , Bertagnolli Mariane
Published: September 2019

Background:
Thrombospondin-1 (Thbs1) is an anti-angiogenic focal adhesion glycoprotein. It binds to receptors CD36, LRP1 and CD47 activating anti-proliferative and anti-migratory mechanisms. Pre-eclampsia (PE) is characterized by poor placenta perfusion and angiogenesis, which is stimulated by oxidant and endoplasmic reticulum (ER) stresses contributing to placental insufficiency. ER stress has also been linked to Thbs1 anti-angiogenic actions. However, whether this mechanism is active in PE is unclear. Our aim was to describe ER stress and Thbs1 regulation in placental tissues and endothelial cells in a genetic mice model of chronic maternal hypertension and superposed pre-eclampsia.

Methods and Results:
Placentas (n=8/group) were collected at 18.5 gestational days from transgenic mice overexpressing human renin and angiotensinogen versus controls (C57BL/6). CD31+ endothelial cells were isolated from placental tissues by magnetic separation. ER stress proteins IRE1a and calreticulin (CLR), as well as Thbs1 and receptors expressions were measured by western blot in placental tissues. CLR and Thbs1 localization in endothelial cells was assessed by immunofluorescence. Data are presented as average±SD. Our results show that IRE1a expression was increased in placentas of hypertensive mice (54±24% P=.05) while CLR was not different. However, in endothelial cells, CLR was shown mobilized from the ER compartment towards the membrane in cells of hypertensive pregnancies or in those exposed to hypoxia. Thbs1 (122±51% P=.03) and receptors CD47 (51±21% P=.03) and LRP1 (58±25% P=.04) were more expressed in placentas of hypertensive pregnancies versus controls, while CD36 was not different. The treatment of endothelial cells of hypertensive pregnancies with a Thbs1 inhibitor, LSKL, has significantly enhanced their angiogenic capacity allowing cells to form closed tubes on matrigel in comparison with the lack of closed tubes in untreated cells.

Conclusion:
Our results described an ER stress state and Thbs1 upregulation in placental tissue and endothelial cells of hypertensive pregnancy. Our findings suggest an important anti-angiogenic role of the Thbs1 mechanism in placentas of hypertensive mice.
Abstract P105: Sex Hormonal Changes of Calcium-Sensing Receptor and Renin-Angiotensin-Aldosterone System in Salt-Sensitive Rats via Exposure of High Salt Diets
Authors: Okechukwu Charles
Published: September 2019

The CaSR regulates Ca
2+
-mediated release of parathyroid hormone (PTH) from parathyroid cells by sensing small changes in serum Ca
2+
concentration. The receptor is also involved in the regulation of cell Ca
2+
levels independent of PTH secretion. The renin-angiotensin-aldosterone system (RAAS) plays vital roles in the development of hypertension, which is associated with vascular and renal dysfunction, and treatment is difficult because the mechanisms are not fully understood. Studies show that adequate Ca
2+
intake (1.0 to 1.5 g/day) is critical for optimal BP regulation and randomized controlled trials have revealed significant reductions in hypertension risk and BP levels in humans. In the kidney, the CaSR is expressed along the nephron and plays an important role, with the Na
+
/Ca
2+
exchanger (NCX), in the regulation of tubular Ca
2+
, Na
+
and water reabsorption. In the present study, we analyzed the effects of mutation, sex hormones and high salt diets on the expression of the CaSR, its downstream signaling complements and proteins in the RAAS in both female and male salt-sensitive (SS) rats. Our data indicate that CaSR gene mutation and high salt diets modulate the expression of the CaSR, GRK2, PKCα, NCX1 and RAAS proteins in the kidney of SS female rats than male rats. And also showed statistically significant difference in electrolytes as shown in the data below. Thus, CaSR signaling is linked to RAAS to control hypertension.
Abstract P106: Elevated Fasting Glucose and Proteinuria in Offspring Born to Gestational Hypertensive African Green Monkeys
Authors: Weaver Chelsea C , Taylor Jessica C , Osborn Jeffrey L
Published: September 2019

Preeclampsia (PreE), or gestational hypertension (GH) concurrent with proteinuria, is a leading cause of maternal and fetal mortality worldwide. PreE often leads to fetal growth restriction resulting in low birth weight offspring. Low birth weight infants are at higher risk of developing cardiovascular disease later in life. The African Green Monkey (AGM;
Chlorocebus aethiops sabaeus
) develops spontaneous GH with proteinuria and fetal growth restriction. We hypothesize that GH will developmentally program offspring to reduced kidney function and metabolic disease. Maternal systolic arterial pressure (SAP) was measured via forearm plethysmography prior to pregnancy and during the 3
rd
trimester of pregnancy. At 1-3 years of age, offspring were single-housed for water intake and urine flow rate collections followed by blood pressure measurement and venous blood sampling. Animals were grouped by pregnancy phenotype. SAP for normotensive (NT, n=23) mothers did not change in pregnancy, but increased during third trimester for GH (Δ=35 ± 7 mmHg, n=34). NT pregnancies resulted in a 4% stillbirth rate (1/23) compared with 29% in GH (10/34). Surviving GH offspring had reduced birth weight (NT 336 ± 11g vs. GH 295 ± 8g, p<0.05). Fasting glucose was higher in progeny born to GH pregnancies (NT 69 ± 8 mg/dl, n=13; GH 92 ± 6 mg/dl, n=20). Urinary protein excretion was higher in offspring born to GH pregnancies (NT 152 ± 27 mg/day; GH 318 ± 46 mg/day). SAP of offspring was similar between maternal phenotype (NT 107 ± 8 mmHg; GH 109 ± 5 mmHg). Juvenile SAP was unrelated to fasting glucose or urinary protein excretion, indicating that these factors are determined by maternal SAP during pregnancy, rather than juvenile phenotype. Thus, GH is associated with fetal growth restriction in the form of high stillbirth rates and lower surviving-offspring birth weights in the AGM. GH offspring have elevated fasting glucose and possible decreased kidney function as evidenced by proteinuria. GH could be programming offspring for early life metabolic risk factors and declining renal function. Future studies include oral glucose tolerance tests and additional markers of renal dysfunction to further assess
in utero
programming of the pathogenesis of cardiovascular and metabolic disease.
Abstract P110: Classical Dendritic Cells Mediate Hypertension by Promoting Oxidative Stress and Renal Fluid Retention
Authors: Lu Xiaohan , Rudemiller Nathan , Privratsky Jamie , Ren Jiafa , Wen Yi , Griffiths Robert , Gunn Michael D , Crowley Steven D
Published: September 2019

Dendritic cells (DCs) promote hypertension via the activation of T cells, but the specific DC subtypes that mediate hypertension require elucidation. We previously found that Fms-like tyrosine kinase 3 ligand-deficient (FLT3L
-/-
) mice that lack classical CD11c
hi
MHCII
hi
DCs have blunted renal accumulation of effector T cells vs wild-type (WT) mice during chronic angiotensin (Ang) II infusion (500ng/kg/min). In the current studies, we tested the hypothesis that classical DCs drive blood pressure elevation by promoting renal fluid and sodium retention. We found that FLT3L
-/-
mice had mean arterial pressures (MAPs) similar to WTs at baseline but had blunted hypertensive responses measured by radiotelemetry during 4 weeks of chronic Ang II infusion (166±2 vs.178±4 mmHg,
p
<0.02). Consistent with their lower blood pressures, the Ang II-infused FLT3L
-/-
mice had attenuated cardiac hypertrophy (6.7±0.1 vs. 7.8±0.4 mg/g body weight,
p
<0.01) and reduced renal mRNA expression for the pro-hypertensive cytokines IL-1β (0.4±0.1 vs. 1.0±0.2; p<0.02) and TNF-α (0.6±0.1 vs. 1.0±0.1; p<0.02). The Ang II-infused FLT3L
-/-
mice had lower urinary excretion of the oxidative stress marker 8-isoprostane (1403±158 vs. 2257±372 pg/24 hours; p<0.04). To examine the effects of classical DCs on renal fluid retention after 7 days of Ang II, we performed an IP saline challenge test. FLT3L
-/-
mice excreted higher proportions of the injected volume (69±7 vs. 49±5%; p<0.04) and sodium (54±7 vs. 38±4%; p=0.07) compared to WTs. Consistent with this enhanced diuresis, mRNA expression for the NCC sodium transporter and all 3 subunits of ENaC were diminished by > 40% (0.4 of 1) in FLT3L
-/-
kidneys compared to the WTs during hypertension. Thus, classical FLT3L
+
DCs promote renal T cell activation with consequent oxidative stress, fluid retention, and blood pressure elevation.
Abstract P1100: Angiotensin-(1-7) Via Angiotensin Type-1 Receptor Synergies Angiotensin Type-2 Receptor Response of Nitrite Formation in Kidney Cells
Authors: Patel Sanket N , Hussain Tahir
Published: September 2019

Angiotensin-(1-7) is evidently considered as an endogenous agonist for Mas receptor. Our recent studies show that MasR and AT
2
R physically interact and in response to their respective agonists Ang-(1-7) and C21 produce interdependent functional response in terms of nitric oxide (NO) generation and natriuresis. Moreover, recent studies suggest that Ang-(1-7) is cardio-protective via AT
1
R stimulation. Considering the complex nature of Ang-(1-7) function, we tested as to how Ang-(1-7) affects AT
2
R function utilizing human kidney proximal tubule epithelial cells (HK-2 cells). Ang-(1-7), C21 or AVE0991 (a MasR selective agonist), alone significantly stimulated NO formation at 1-10 μM (50-100%, n=3), but not at lower concentrations (0.1-100 nM). In the interaction studies, the non-responsive concentrations (0.1-10 nM) of Ang-(1-7) and C21 together produced a synergy in producing NO in HK-2 cells (p<0.05 vs. either ligand alone). The synergy is observed only when the Ang-(1-7) was added to the HK-2 cells 10 min prior to the C21, not if the agonists were added simultaneously. The synergy between Ang-(1-7) and C21 was sensitive to AT
2
R antagonist (PD123319, 10 μM) and AT
1
R antagonist (candesartan, 10 μM), but not MasR antagonist (A779, 10 μM). Interestingly, pre-stimulation with synthetic non-peptide MasR selective agonist AVE0991 at any concentrations (0.1-100 nM) did not show synergism with AT
2
R agonist C21. Collectively, results show Ang-(1-7) at lower concentrations via AT
1
R, not MasR, synergies the AT
2
R function and this phenomenon may have physiological significance.
Abstract P1103: Angiotensin II Type I Receptor Agonistic Autoantibody Blockade Improves Hypertension and Immune Activation in Postpartum Preeclamptic Rats
Authors: Cunningham Mark W , Chinthakuntla Pooja , Brooks Karen , Fisher Brandon , Ibrahim Tarek , Amaral Lorena , Booz George W
Published: September 2019

Women with preeclampsia (PE) have a risk of developing cardiovascular diseases (CVD) later in life. The angiotensin II type I receptor agonistic autoantibodies (AT1-AAs) are elevated in women with PE, PE women 2 years post-partum (PP), and the reduced uterine perfusion pressure (RUPP) rat model of PE. Blockade of the AT1-AA by using a specific binding seven amino acid peptide sequence (7AA) improves the pathology of PE in RUPP rats. The long-term effects of AT1-AA inhibition on blood pressure, NK cell activation, heart mitochondria (mt) proteins, and CVD in the RUPP model PP is unknown. Therefore, we hypothesized that PP RUPP rats have elevated blood pressure, NK cell activation, and changes in heart mt proteins, which will be prevented in RUPP rats administered the 7AA during pregnancy.

Methods:
Pregnant Sprague Dawley rats were divided into groups; normal pregnant (NP) (n=7), RUPP (n=10), and RUPP+7AA (n=9). Gestational day 14, RUPP surgery was performed and 7AA (2 μg/ml) administered via minipump.

Results:
At 8 and 10 weeks (wks) PP, blood pressure (MAP), blood, and hearts were collected. NK cells were quantified by flow cytometry. At 8 wks PP, MAP was elevated in RUPP vs. NP (130±2 vs. 123±4 mmHg, ns), and RUPP+7AA (124±4 mmHg) treatment prevented this increase. At 10 wks, MAP was elevated in RUPP vs NP (133±5 vs. 120±5 mmHg, p=0.08), with a significant decrease in MAP in RUPP+7AA (107±6 mmHg) vs. RUPP (p<0.05). Total circulating NK cells were increased in RUPP vs NP (45±9. vs. 29± 8% gated cells, ns), which was prevented in RUPP+7AA (19±16 % gated cells) at 8 wks PP. Hearts were enlarged with RUPP vs NP (0.41±0.04 vs. 0.36±0.02g/100gBW, ns), which was normalized in RUPP+7AA (0.34±0.02g/100gBW). Previous studies show that during pregnancy complex IV is significantly lower along with a decrease mt function in RUPP vs NP. Complex IV mt proteins in the heart were elevated in RUPP+7AA vs. RUPP (5.5±1.7 vs. 3.0±0.2 AU, ns).

Conclusion:
In summary, PP PE rats have an increase in MAP, NK cells, and larger hearts. AT1-AA inhibition restores complex IV mt levels and improves HTN, immune activation, and cardiac hypertrophy PP. This study highlights the importance of AT1-AA inhibition during PE to prevent CVD later in life.
Supported: AHA18CDA34110264
Abstract P1106: RND3/rhoe Deficiency Results in Mitochondrial Dysfunction in Placenta With Preeclampsia Through Modulating PPARγ Pathway
Authors: Yue Xiaojing , Yang Xiangsheng , Ma Yanlin , Chen Xia , Huang Liping , Yu Yanhong
Published: September 2019

Preeclampsia (PE) is a life-threaten disease among the pregnant women with the syndromes of severe hypertension, proteinuria and pitting edema. Mitochondrial dysfunction mediated oxidative stress has been implicated in the placentae with PE. However, the regulatory mechanism causing the mitochondrial dysfunction is still unknown. In this study, we identified Rnd3, a small Rho GTPase, as a new factor participating in the regulation of mitochondrial function. 46% and 51% decreases in RND3 expression levels were found in the human placentae with PE and in the primary trophoblasts isolated from PE patients, respectively. The downregulation of RND3 in the human PE primary trophoblasts was associated with excessive ROS generation and damaged mitochondrial integrity. To explore the role of Rnd3 in mitochondria, we knocked down of RND3 in trophoblastic BeWo cells and detected the loss of mitochondria cristae, the generation of mitochondrial dygradosome, the mitochondrial membrane potential depolarization and the 4.9 fold increased proton leak from mitochondrial respiratory chain. The overexpression of GFP-Rnd3, by contrast, attenuated hypoxia induced oxidative stress and apoptosis in BeWo cells. In the mechanistic studies, up-regulation of Rnd3 resulted in the nuclear accumulation of transcription factor PPARγ. By stimulation of PPARγ in BeWo cells, mitochondrial uncoupling protein 1 (UCP1) cascade was activated, and the Rnd3 deficiency mediated mitochondrial proton leak was attenuated. Finally, we performed the adenovirus mediated Rnd3 overexpression in the human PE primary trophoblasts, and detected significant decreases in ROS level and apoptosis. We concluded that Rnd3 acted as a novel protective factor in placental mitochondria through PPARγ-UCP1 signaling promotion. Rnd3 downregulation observed in PE patients may explain one of the pathogeneses of severe placental oxidative stress.
Abstract P1108: Serum Immunoglobulins are Not Necessary for Experimental Hypertension
Authors: Chen Yuhan , Dale Bethany , Alexander Matthew , Smart Duncan , Ao Mingfang , Madhur Meena S
Published: September 2019

The role of B cells and serum immunoglobulins (Ig) in hypertension is unclear. Elevated serum IgG has been observed in some hypertensive patients and in animal models of hypertension. Mice with a severe B cell deficiency due to lack of B cell activating factor receptor (BAFF-R) and mice lacking the cytokine interleukin 21 (IL21) - which is a potent inducer of B cell Ig class switching and high affinity Ig production - are protected from Ang II-induced hypertension and exhibit reduced serum IgG levels. Thus, we sought to determine whether hypertensive IgGs play a causative role in hypertension and end-organ dysfunction. First, we purified serum IgG from normotensive and hypertensive wild type (WT) mice and adoptively transferred equal amounts of IgG by intraperitoneal injection into normotensive recipient mice followed by infusion of a sub-pressor dose of angiotensin II (Ang II) for 2 weeks. Flow cytometric analysis of aortas revealed a modest increase in F4/80+ macrophages (p=0.04) and CD8+ T cells (p=0.02) in mice that received hypertensive IgG. However, there was no difference in endothelial dependent relaxation, renal inflammation, albuminuria, and blood pressure (BP) between the two groups. As an alternative method to investigate the role of class-switched high affinity Igs, we studied mice with genetic deletion of activation-induced deaminase (AID) which is an enzyme necessary for somatic hypermutation (to generate high affinity Igs) and isotype class switching. AID
-/-
mice exhibit undetectable IgG and higher levels of IgM in serum compared to WT mice. Ang II infusion for 4 weeks revealed no difference in BP, aortic inflammation, renal inflammation, and albuminuria. We also studied mice with deficiency of the IgM heavy chain (uMT). These mice exhibit undetectable levels of all immunoglobulins and are severely deficient in mature B cells. uMT-/- mice are also not protected from hypertension induced by Ang II infusion or deoxycorticosterone acetate (DOCA-salt) treatment. Together, these results demonstrate that while elevated serum immunoglobulins seen in hypertensive animals and humans may represent a biomarker of aberrant immune activation, they are likely not playing a causal role in hypertension pathophysiology.
Abstract P1110: Tumor Necrosis Factor Alpha Blockade Improves Natural Killer Cell Activation, Hypertension, and Mitochondrial Oxidative Stress in a Preclinical Rat Model of Preeclampsia
Authors: Cunningham Mark W , Jayaram Aswathi , Amaral Lorena A , Chinthakuntla Pooja , Brooks Karen , Booz George , Vaka Venkata R , Ibrahim Tarek , Deer Evangeline , Cornelius Denise C , LaMarca Babbette
Published: September 2019

Preeclamptic (PE) women have placental ischemia, hypertension (HTN), mitochondrial (mt) dysfunction, increased mt-reactive oxygen species (ROS), cytolytic natural killer (NK) cells, and pro-inflammatory cytokines, such as tumor necrosis factor (TNFα). The reduced uterine perfusion pressure (RUPP) preclinical rat model of PE, is a well-established model to study mechanisms of hypertension in response to placental ischemia during pregnancy. Previous studies from our lab have shown that TNFα is elevated in RUPP rats and that blockade of TNFα, via etanercept (ETAN) decreases TNFα and HTN. However, the effect of TNFα blockade on NK cell activation, mt protein/function, and mtROS is unknown. We hypothesized that ETAN would decrease blood pressure, NK cell activation, mtROS, and improve mt function in RUPP rats. Rats were divided into 4 groups: normal pregnant (NP) (n=41), RUPP (n=34), RUPP+low dose (LD) ETAN (0.4 mg/kg) (n=11), RUPP+high dose (HD) ETAN (0.8 mg/kg) (n=19). LD was used previously, while HD is clinically used to treat chronic inflammatory diseases. Gestational day 14, RUPP surgery was performed and ETAN administered subcutaneous on day 18. Day 19, conscious blood pressure (MAP), blood and tissues were collected, and mt were isolated. Flow cytometry was used for quantification of NK cells. MAP was elevated in RUPP vs. NP (119 ± 1 vs. 102± 1 mmHg, p<0.05) which was reduced to 110 ± 2 in RUPP + LD ETAN (p<0.05 vs. RUPP) and 116 ± 1 in RUPP + HD ETAN. Activated NK cells were increased in circulation (5.9 ± 1.8 vs. 4.0 ± 1.4 %), placenta (4.8 ± 1.3 vs. 1.9 ± 0.7 %), and kidneys (3.1 ± 0.5 vs. 0.8 ± 0.5%) of RUPP vs. NP (p<0.05). Both LD (2.1 ± 0.7%, 1.7 ± 0.7%, 0.8 ± 0.5%) and HD (0.2 ± 0.1%, 0.4 ± 0.1%, 0.8 ± 0.2%) ETAN decreased activated NK cells in circulation, placenta, and kidney respectively (p<0.05 vs RUPP). Placental (0.79± 0.02 vs 1±0.05 fold) and renal (0.67± 0.05 vs 1±0.06 fold) mtROS was reduced with LD ETAN vs RUPP (p<0.05). ETAN treatment lowers blood pressure and reduces circulating, placental, and renal activated NK cells and mtROS in placental ischemic rats. Therefore ETAN could be a potential therapeutic for improving HTN, inflammation, and mitochondrial function during complicated pregnancies.
Supported: AHA18CDA34110264, HL130456, R01HD067541, & P20GM121334
Abstract P1112: Gestational Interventions and Fetal Outcomes in a Rat Model of Intrauterine Growth Restriction
Authors: Coats Laura E , Davis Gwendolyn K , Bamrick Daniel R , Bakrania Bhavisha A , Ojeda Norma B , Alexander Barbara T
Published: September 2019

Preeclampsia (PE) is a leading cause of maternal and fetal mortality worldwide. Numerous studies indicate blood pressure is increased in individuals exposed to PE during fetal life. Currently, there are no effective therapeutics for PE. Our laboratory utilizes a rat model of PE induced by placental ischemia, the reduced uterine perfusion pressure (RUPP) model that results in intrauterine growth restriction (IUGR) and hypertension in the offspring. IUGR is dictated by inadequate uteroplacental blood flow and insufficient nutrient transport to the fetus. Preclinical use of a soluble guanylate cyclase (sGC) inducer abolishes hypertension in RUPP dams, but its effect on offspring is unknown. The purpose of this study was to test the hypothesis that maternal administration of a sGC inducer improves placental perfusion and nutrient transport, mitigating IUGR. RUPP or sham procedure was performed at day 14 of gestation and dams were administered vehicle or sGC inducer (riociguat, 10mg/kg/day, sc) from days 14 to 20 gestation. At day 20 of gestation uterine artery resistance index (UARI) was increased, indicative of poor perfusion, in vehicle-treated RUPP compared to vehicle-treated Sham (0.64±0.01 vs. 0.57±0.01 mm/s, respectively;
p
<0.05); UARI was not significantly increased in sGC-treated RUPP (0.59±0.02;
p
<0.05 mm/s). Fetal pup weight was reduced in vehicle-treated RUPP compared to vehicle-treated Sham (3.74±0.10 vs. 4.19±0.08 g, respectively;
p
<0.05); yet with treatment, fetal pup weight was not decreased. Heart fatty acid binding protein (hFABP) protein expression is increased in women with PE, a means to maintain normal brain development in IUGR offspring. This increase was observed in vehicle-treated RUPP compared to vehicle-treated Sham (1004±32 vs. 583±65 A.U., respectively,
p
<0.05). After treatment, placental hFABP protein expression was increased in sGC-treated Sham compared to vehicle-treated Sham (987±95 vs. 583±65 A.U., respectively;
p
<0.05), but was reduced in sGC-treated RUPP compared to vehicle-treated RUPP (732±54 vs. 1004±32 A.U., respectively;
p
<0.05). Further studies are warranted to determine the long-term benefit of fetal exposure to a sGC inducer and its effect on blood pressure and long term health in the offspring.
Abstract P1116: Early Life Stress Induces HDAC-Dependent Expression of Pro-Oxidant Genes in the Aorta
Authors: Neder Thomas H , Hyndman Kelly A , Ho Dao H , Zarzour Abdalrahman , Johnson Alan K , Weintraub Neal , Pollock David M , Pollock Jennifer S
Published: September 2019

Childhood adversity or early life stress (ELS) is an understudied risk factor for cardiovascular disease (CVD). The mouse ELS model, maternal separation with early weaning (MSEW), generates robust effects on anxiety as well as immunological consequences in adulthood. Previous studies from our laboratory showed that MSEW induces a pro-oxidant vascular phenotype in adulthood. Early in life or during development, epigenetic mechanisms such as the cellular balance of histone acetylation and deacetylation (HDACs) critically influences gene expression. We hypothesized that MSEW induces HDAC-dependent expression of pro-oxidant genes in the vasculature. Thoracic aortae were isolated from male adult (12 wk) normally-reared (NR) or MSEW mice. Histone 3 lysine acetylation (H3K27) status was found to be significantly decreased in aorta from MSEW compared to NR mice, indicating MSEW induced increased HDAC activity. The MSEW-induced increased in aortic NADPH oxidase (NOX) 2, NOX4, and toll-like receptor (TLR) 4 expression and was significantly blunted with
ex vivo
trichostatin A (TSA) treatment with no changes in NR mice (relative mRNA; NOX2: MSEW+VEH 1.55±0.12 vs MSEW+TSA 1.24±0.16, NOX4: MSEW+VEH 1.68±0.14 vs MSEW+TSA 1.25±0.07, TLR4: MSEW+VEH 1.65±0.16 vs MSEW+TSA 0.79±0.09, N=4-8, p<0.05). Further, MSEW-induced endothelial dysfunction was reversed with
ex vivo
TSA treatment with no changes observed in NR mice (E
max
NR+VEH 86.3±3.5, NR+TSA 82.8±6.3, MSEW+VEH 64.1±8.2 vs MSEW+TSA 85.9±1.7, N=8-12, p<0.05). Reports show that HDAC9 is associated with increased risk for vascular diseases. Aorta from MSEW mice had a significant increase in HDAC9 abundance compared to NR mice (rel. mRNA; NR: 1.02±0.1 vs MSEW: 1.67±0.2, N=6, p<0.05). To determine whether HDAC9 directly induces NOX2, NOX4, and/or TLR4 expression, we utilized HDAC9 over-expressing (HDAC9-OE) mice. Aortic tissue from HDAC9-OE mice revealed a 10-fold NOX4 and 13-fold TLR4 increase, with no change in NOX2 expression. Our results indicate that HDACs, especially HDAC9, are ELS-sensitive mediator(s) of pro-oxidant gene expression. We conclude, that activation of HDAC-dependent pro-oxidant genes, specifically NOX4 and TLR4 pathways, contribute to increased CVD risk following ELS.
Abstract P112: Sex Differences and the Role of G Protein-Coupled Estrogen Receptor in Arterial Stiffening
Authors: Ogola Benard O , Clark Gabrielle L , Abshire Caleb M , Harris Nicholas R , Gentry Kaylee L , Lawrence Dylan , Zimmerman Margaret A , Bayer Carolyn L , Miller Kristin S , Lindsey Sarah H
Published: September 2019

Pulse wave velocity (PWV) independently predicts cardiovascular events and is exacerbated in women following menopause. Since our previous work shows that G protein-coupled estrogen receptor (GPER) plays a protective role in the vasculature, the current study evaluated sex differences and the impact of GPER on arterial stiffening. We hypothesized that genetic deletion of GPER attenuates sex differences in arterial stiffness. Male and female wildtype (wt) and global GPER knockout (ko) mice (n=46) were used between 16-21 weeks of age. Ang II infusion (700 ng/kg/day for two weeks) was used to induce hypertension, and SBP was measured using tail cuff plethysmography. Local PWV within the carotid artery was obtained via high frequency ultrasound in both color Doppler and M-mode. The excised carotid was subjected to passive biaxial mechanical testing followed by four fiber family constitutive modeling. Statistical analysis was performed using two-way ANOVA with Sidak’s multiple comparisons test. Baseline SBP was significantly lower in females (P=0.035) but was not impacted by genotype. Baseline PWV was significantly higher in male versus female wt mice (P<0.01) but was not different in ko mice. Baseline axial (P<0.01) and circumferential (P=0.027) stiffness was higher in female wt mice but was not impacted by genotype. Ang II infusion significantly increased SBP (P<0.001) and PWV (P<0.001) in all groups, removing any impact of sex or genotype. Ang II increased axial stiffness and decreased diagonal collagen in male and female GPER ko mice. In conclusion, we found that GPER deletion did not impact blood pressure but removed sex differences in PWV. While female wt mice had the lowest PWV at baseline, genetic deletion of GPER or Ang II infusion removed this protection independent of BP. Therefore, local carotid PWV may provide information on vascular status independent of BP, and GPER plays an important role in vascular compliance and arterial stiffening.
Abstract P1120: Opioids Induce Proliferation in Vascular Smooth Muscle Cells Leading to an Increase in Myogenic Tone in Resistance Arteries
Authors: Tomcho Jeremy C , Edwards Jonnelle , Bearss Nicole R , McCarthy Cameron G , Joe Bina , Wenceslau Camilla F
Published: September 2019

The United States of America is currently undergoing an opioid crisis. According to the 2018 CDC Annual Surveillance Report of Drug Related Risks and Outcomes there were 78,840 hospitalizations and an estimated 140,077 emergency room visits for opioid related poisonings in 2015 alone. It has been shown that opioid exposure increases vascular age relative to chronological age. Further, long-term abuse of opioids leads to changes in regional cerebral blood flow. However, there is a lack of understanding about acute and chronic opioid abuse and how it affects microvasculature structure and function; specifically on myogenic tone, a fundamental mechanism to maintain blood flow to important tissues including the brain, kidneys, and more. We hypothesize that opioids cause an increase in proliferation of vascular smooth muscle cells (VSMC) leading to an increase in myogenic tone in mesenteric resistance arteries. To test this hypothesis, aortic vascular smooth muscle cells were cultured and treated with the endogenous opioid (ENO) beta-endorphin or the exogenous opioid (EXO) morphine. ENO treatment increased cellular proliferation with chronic exposure (greater than 72 hours). We measured a maximum proliferation at 120 hours and performed 2 way ANOVA with Bonferroni posttests [confluence (%): DMSO vehicle 20.4 ± 4.8, 1 nM 34.9 ± 7.4*, and 1 μM 29.9 ± 7.0; * p<0.05]. Increased proliferation was also demonstrated in cells treated with EXO. We measured a maximum proliferation at 120 hours and performed 2 way ANOVA with Bonferroni posttests [confluence (%): DMSO vehicle 20.4 ± 4.8, 1 nM 32.6 ± 9.0, 1μM 33.3 ± 10.2*, and 10 μM 29.7 ± 4.4; * p<0.05]. We also observed that when acutely exposing mesenteric resistance arteries to EXO (1.0 μM) there was a left shift in myogenic tone to 80 mmHg as opposed to 100 mmHg in the in absence of EXO. In conclusion, our results suggest that chronic exposure to ENO and EXO increase VSMC proliferation. Further, acute exposure to EXO was able to increase myogenic tone in resistance arteries. This suggests that opioid dependence could contribute to the development of peripheral vascular disorders such as hypertension.
Abstract P1124: Nephron Deficiency and Hyperglycemic Renal Injury Using a Novel One Kidney Rat Model
Authors: Cobb Meredith B , Wu Wenjie , Johnson Ashley C , Attipoe Esinam M , Garrett Michael R
Published: September 2019

Patients with hypertension or diabetes are at higher risk for developing chronic kidney disease (CKD). Low nephron number has been linked with susceptibility to develop hypertension and CKD, but a clear connection between nephron number and hyperglycemic renal injury has not been established. Thus, we seek to investigate the association between nephron deficiency and the development of diabetic kidney disease using a unique model of nephron deficiency, the HSRA rat. Offspring of HSRA are born with a single kidney 50-75% of the time with the remaining pups born with two kidneys. This provides the advantage of being able to directly compare congenital one-kidney, nephron-deficient rats (HSRA-S, ~20,400 nephrons) with nephrectomized two-kidney rats (HSRA-UNX, ~25,100 nephrons) and two-kidney control rats (HSRA-C, ~50,000 nephrons). Previous work has shown that HSRA-S develops increased renal dysfunction with age compared to HSRA-UNX and HSRA-C, which is greatly exacerbated in the presence of DOCA hypertension. Our hypothesis is that a secondary stressor of hyperglycemia in the context of low nephron number will similarly cause greater decline in renal function in HSRA-S compared to HSRA-UNX and HSRA-C. Streptozotocin (STZ) was administered at 9 weeks of age in all three groups and animals were followed until week 24. Despite overt hyperglycemia (350-450 mg/dl), diabetic groups did not develop increased proteinuria compared to their non-diabetic counterparts. Notably, a significant increase in kidney weight was observed in HSRA-S+STZ compared to HSRA-S (kidney weight to body weight ratio [mg/g] 4.8 ± 0.3089 for HSRA-S vs 7.8 ± 1.204 for HSRA-S+STZ, p=0.02), suggesting that hyperglycemia has a deleterious impact on kidney function via hyperfiltration and hypertrophy.
Despite no significant effect on proteinuria after an “early insult,” we are investigating the impact of hyperglycemia after overt injury (“late insult”) is observed in HSRA-S (week >24). We will also revisit an “early insult” of hyperglycemia with the addition of modest hypertension (130 mmHg) via DOCA, which represents patients with both diabetes and hypertension
. Through this research, we hope to contribute to the growing knowledge of the relationship between nephron number and disease.