bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 090: Increased Uterine Artery Resistance Index Results In Placental Hypoxia And Tnf-α In The Dahl Salt Sensitive Rat During Pregnancy
Authors: Gillis Ellen E , Garrett Michael R , Sasser Jennifer M
Published: March 2022

In normal pregnancy, uterine artery resistance index (UARI) decreases as pregnancy progresses to accommodate the increased blood supply to the growing fetus. In preeclampsia, increased UARI impedes the utero-placental blood flow. Recent studies in our lab indicate that the Dahl salt sensitive (S) rat spontaneously develops a preeclamptic phenotype, characterized by increased blood pressure and proteinuria as pregnancy progresses and a decrease in litter and pup size. Here, we tested the hypothesis that the phenotype in the Dahl S rat is due to decreased placental blood flow, causing the placenta to become hypoxic, resulting in increased production of TNF-α. Female Dahl S and Sprague Dawley rats were mated with males of their same strain, and pregnancy was confirmed by presence of sperm (D1). UARI was measured during mid and late pregnancy (D14 and 18, respectively) via Doppler ultrasound in anesthetized animals (n= 4-12). Rats were sacrificed on D20, and placentas were harvested for further analysis. TNF-α and HIF-1α were measured in homogenized placentas via commercially available ELISA (n= 5) and Western Blot analysis (n= 9-11), respectively. UARI decreased in the Sprague Dawley rat between D14-18, but increased in the Dahl S rat (Table). HIF-1α was significantly increased in the Dahl S placenta vs Sprague Dawley (Table), and TNF-α also showed a tendency to increase in the Dahl S placenta (p=0.08). These data suggest that the relative decrease in blood flow to the placenta during pregnancy in the Dahl S rat causes a hypoxic environment, leading to the release of vasoactive factors that contribute to the development of the preeclamptic phenotype observed in this strain.
Abstract 093: Progesterone Supplementation Improves Blood Pressure And Uterine Artery Resistance In Response To Placental Ischemia During Pregnancy
Authors: Amaral Lorena M , Moseley Janae , Cornelius Denise C , Martin James N , LaMarca Babbette
Published: March 2022

Preeclampsia (PE), new onset hypertension at 20 weeks of gestation, is characterized by increased uterine artery resistance index (UARI), chronic immune activation and decreased of vasodilators such as nitric oxide (NO). Despite being one of the leading causes of death in pregnant women, currently there is no effective treatment for PE except for early delivery of the fetus. We have demonstrated that PE women have significantly lower circulating progesterone than normal pregnant (NP). 17-alpha-hydroxyprogesterone caproate (17-OHPC) is a synthetic metabolite of progesterone used for the prevention of recurrent preterm birth and although the mechanism is not understood, is suspected to have vasodilatory and anti-inflammatory effects. Therefore, we hypothesized that progesterone (17-OHPC) supplementation could reduce blood pressure (MAP), pro-inflammatory cytokines, CD4+ T cells, UARI, as well as increase NO bioavailability in a hypertensive rat model of PE. To address this question 17-OHPC (3.32mg/kg) was intraperitoneally administered on day 18 of gestation into Reduced Uterine Perfusion Pressure (RUPP) rats and carotid catheters were inserted. MAP, blood and tissues were collected on day 19. MAP in NP rats (n=13) was 92±2; 123±2 in RUPP (n=18), and 116 ±1 mmHg in RUPP+17-OHPC (n=10), p <0.05. UARI was 0.78±0.03 in RUPP (n=4) and 0.63±0.038 in RUPP+17-OHPC (n=8), p<0.05. Circulating CD4+ T cells were 1.19±1.0% of gated cells in NP (n=7), which increased to 8.52±2.4% in RUPP rats (n=10) but was significantly reduced to 2.72±0.87% (n=14) in RUPP + 17 O-HPC, p <0.05. Total circulating nitrate/nitrite was 26.34 ±3.5 μM in NP (n=12); 14.58±3.1 in RUPP rats (n=8) and increased to 26.69±1.62 in RUPP+17-OHPC (n=7), p <0.05. Aortic eNOS expression was 0.65±0.11 A.U in NP (n=4), which decreased to 0.33±0.01 in RUPP rats (n=4) but increased to 0.57±0.01 in RUPP+17-OHPC (n=5), p <0.05. Levels of TNF-alpha were 65.84±17.7 pg/ml in RUPP rats (n=5) but were blunted to 17.24±3.9 in RUPP+17-OHPC (n=8), p <0.05. In conclusion, 17-OHPC supplementation improves inflammation, UARI, hypertension, and nitric oxide bioavailability in response to placental ischemia during pregnancy and should therefore be considered further for addition to the clinical management of PE
Abstract 096: Secreted miR-27a in Extracellular Vesicles from THP-1 Cells Cause Hypertension by Reduced Angiotensin-(1-7)-Mas Receptor Expression and Function in Artery
Authors: Zou Xue , Wang Jialiang , Zeng Chunyu
Published: March 2022

Extracellular vesicles (EVs) play important role in various diseases. Essential hypertension is found to be with higher plasma EV concentrations, whether or not the EVs are involved into pathogenesis of hypertension is not known. Our present study found injection of EVs from THP-1 cells, a monocyte cell line, to SD rats increased blood pressure, accompanied with impaired Ang 1-7 mediated vasodilation in mesenteric artery preconstricted with norepinephrine. In EVs treated SD rats, Mas expression and eNOs expression were decreased in mesenteric artery. After treatment of THP-1 cells with LPS, which represents the inflammatory states, LPS-treated EVs worsen Ang 1-7 mediated vasodilative effect, and further increased blood pressure to higher degree. After degradation of RNAs in EVs by Triton X-100 and RNase, the hypertensive effect of EVs was blocked, indicating RNAs play a vital role in this action. Screening studies, including analysis of gene microarray, comparison of plasma microRNA levels between hypertensive and healthy controls, analysis of microRNA changes with LPS, found that miR-27a is a possible candidate. It is interesting to find that the target of miR-27a is Mas, predicted by genetics software. Immunofluorescence study showed EVs could be transferred to the cytosol and nucleus in the receipt cells. Through loss- and gain-of function approaches, we found that miR-27a decreased Mas and eNOS expressions in HUVECs. The transferred miR-27a in EVs is physiological significance, because injection of EVs from miR-27 transfected HEK293 cells increased blood pressure in SD rats, with impaired Ang 1-7 mediated vasodilation preconstricted by norepinephrine. Moreover, the expressions of Mas and eNOS were decreased in the EVs treated rats. Therefore, we conclude that miR-27a in EVs derived from THP-1 cells decreases Mas receptor expression in HUVECs, consequently influences endothelial NO release and impaired Ang 1-7 mediated vasodilative function, which is involved in the pathogenesis of hypertension.
Abstract 201: Overexpression of Calcium/Calmodulin-Dependent Protein Kinase II (CaMKII) Potentiates Angiotensin II (AngII)-Mediated Signaling in Neurons
Authors: BASU URMI , Case Adam J , Liu Jinxu , Li Yu-Long , Zimmerman Matthew C
Published: March 2022

AngII signaling in the brain regulates systemic cardiovascular function. AngII modulates neuronal ion channel activity and neuronal firing via reactive oxygen species (ROS) and redox-sensitive proteins. Our recent studies indicate that in neurons AngII increases activity (i.e. phosphorylation) of CaMKII, a signaling intermediate known to inhibit potassium (K+) channel current. However, the potential cross-talk between AngII, ROS, and CaMKII in modulating neuronal K+ channel activity and the in vivo central AngII-induced pressor response remains unclear. Here, we tested the hypothesis that CaMKII overexpression exacerbates the AngII-dependent inhibition of K+ channel current and the acute central AngII-induced increase in blood pressure. The neuron specific isoform of CaMKII (CaMKIIα) was cloned, and CaMKIIα adenovirus was generated (AdCaMKIIα). Adenovirus-mediated overexpression of total and active CaMKII protein were assessed in differentiated mouse catecholaminergic (CATH.a) neurons by measuring total and phosphorylated CaMKII protein levels, respectively, via Western blot analysis. To investigate the effect of CaMKIIα overexpression on K+ channel activity, CATH.a neurons were transduced with AdCaMKIIα or control adenovirus, AdEmpty, and voltage-gated K+ current (IKV) was measured by the whole cell patch-clamp technique. Baseline IKV was measured followed by 5 mins of AngII (100 nM) superfusion. AngII lowered steady-state current (ISS) and peak current (Ipeak) in AdEmpty-transduced neurons (ISS by 20 ± 4%; Ipeak by 19 ± 3%), and this inhibitory response was significantly potentiated in AdCaMKIIα-transduced neurons (ISS by 40 ± 7%; Ipeak by 41 ± 5%; p< 0.05). Preliminary in vivo studies suggest that the central AngII-induced pressor response is exacerbated in mice intracerebroventricularly injected with AdCaMKIIα as compared to control virus-injected mice. These data indicate that CaMKIIα overexpression potentiates AngII-mediated inhibition of IKv and the central AngII-induced pressor response. Future studies will investigate the ROS-mediated post-translational modifications of CaMKIIα in AngII-stimulated neurons and the contribution of these modifications to ion channel activity and blood pressure regulation.
Abstract 202: Involvement of neuronal high mobility group box 1 (HMGB1) in Angiotensin II-mediated neuronal-microglial interaction
Authors: Song Chunjuan , Santisteban Monica , Zubcevic Jasenka , Raizada Mohan
Published: March 2022

We have demonstrated that activation of microglia in the autonomic brain regions, primarily in the paraventricular nucleus (PVN), is critical in the development and establishment of hypertension. Neuroimmune synaptic communication is implicated in activating microglia and inducing neuroinflammation in many neuropathophysiological diseases. Specifically, HMGB1 secreted by neurons is a critical mediator in neuroimmune crosstalk. Thus we hypothesized that HMGB1 is a key regulator involved in microglia activation in Ang II-induced hypertension. Neuronal and microglial cells in primary culture were established for this study. Effect of Ang II on microglial movement was recorded and quantified by time-lapse microscopy and video analysis. In addition, the expression, translocation and secretion of HMGB1 in neurons were measured by qPCR, Western blot and immunocytochemistry.

Ang II treatment of neuronal-microglial co-culture resulted in a 5-fold increase of microglial migration towards neurons, which was completely abolished by treatment with 1 μM Losartan, an Ang II receptor type 1 antagonist. The presence of neurons was found to be essential for microglial movement since Ang II failed to induce these effects in the absence of neurons. Neurons express high levels of HMGB1, which was predominantly localized in the nucleus. Ang II treatment resulted in a time-dependent translocation of nuclear hMGB1 into the cytoplasm and ultimately into the medium. Both translocation and release of Ang II-induced HMGB1 was blocked by Losartan. Furthermore treatment of microglia with 500ng/ml recombinant HMGB1 caused ~2 fold increase of proinflammatory cytokines (TNF-α and IL-1β). The importance of HMGB1 is further underscored by our observation that its protein level was increased ~2 fold in the PVN but not in the nucleus of the solitary tract (NTS) or rostral ventrolateral medulla (RVLM) of the spontaneously hypertensive rats (SHR) compared with the normotensive WKY rats. Taken together, these observations suggest that Ang II primarily activates the neuronal AT1R to increase the expression, cytosolic translocation and release of HMGB1. The released HMGB1 primes microglial activation and release of cytokines, two key events in neuroinflammation in hypertension.
Abstract 209: Impact Of Hyperandrogenemia On Cardiovascular And Metabolic Function And Ventilatory Responses To Hypercapnia In Female Shrs
Authors: do Carmo Jussara M , da Silva Alexandre A , Hougton Haley J , Moak Sydney P , Smith Andrew , Hall John E
Published: March 2022

Sex hormones influence regulation of blood pressure (BP), metabolism and ventilation. In this study we examined whether chronic hyperandrogenemia, characteristic of polycystic ovary syndrome, alters BP, metabolism and ventilatory responses to hypercapnia in female spontaneously hypertensive rats (SHRs). Female SHRs at 4 weeks of age were implanted subcutaneously with dihydrotestosterone pellets (DHT, 7.5 mg/90 days, n=5). Female untreated SHRs were used as control (n=5). At 15 weeks of age, the rats were implanted with telemetry probes to measure BP and heart rate (HR). After 10 days of recovery from surgery, tidal volume (V
T
), respiratory frequency (f
R
) and pulmonary ventilation (V
E
) were measured by whole-body plethysmography during normocapnia (21% O
2
) or hypercapnia (

7% CO
2

). Baseline mean arterial pressure (MAP), heart rate (HR) and body composition were measured. Compared to untreated controls, DHT treated SHRs were heavier (246±3 vs. 201±5g), with more adiposity (23±2 vs. 4±3 g) and lean mass (206±8 vs. 174±9 g), but similar food intake (15±1 vs. 13±1 g) and MAP (141±2 vs. 142±6 mmHg), whereas HR was significantly lower (332±9 vs. 365±6 bpm). The DHT-treated group also showed
increased
ventilatory response
to hypercania (1541
±161 vs.
1091
±142 ml.min
-1
.kg
-1
), due to increased f
R
(146±5 vs. 133 breaths/min) and V
T
(11±1 vs 8±1 ml/kg). MAP and HR responses to hypercapnia were similar between groups. These results indicate that testosterone increases body weight and ventilatory responses to hypercapnia without changing BP in female SHRs. (NHLBI-PO1HL51971, NIGMS-P20GM104357 and AHA SDG5680016)
Abstract 216: Left Ventricular Diastolic Dyssynchrony In Patients With Treatment-naïVe Hypertension And The Effects Of Antihypertensive Therapy
Authors: Kwon BeomJune , Woong RohJi , Kim HeeYeol , Ihm SangHyun Ihm
Published: March 2022

Backgrounds:
We tried to investigate whether differ in various blood pressure (BP) parameters between the patients with low normal TSH and high normal TSH levels.

Methods:
The study groups consisted of 285 patients with treatment-naïve hypertension and euthryoid undergoing comprehensive echocardiography (TTE), brachial-ankle pulse wave velocity (PWV), ankle-brachial index, central BPs, and ambulatory BP monitoring (ABPM) evaluations. All patients were divided into those with low normal TSH (0.40~1.99 μIU/ml, group 1) or high-normal TSH (2.00-4.50 μIU/ml, group 2).

Results:
Hypertensive male patients were divided into group 1 (n=113, 68.9%) and group 2 (n=51, 31.1%) and female patients were divided into group 1 (n=71, 58.7%) and group 2 (n=50, 41.3%). Multivariate analyses revealed augmentation index (AIx, 71.0 [adjusted mean]±1.7 [standard error] vs. 78.8±2.5 %, p=0.045), central systolic BP (SBP) (143.3±2.1 vs. 153.0±3.2 mmHg, p=0.013), systemic vascular resistance (21.4±0.6 vs. 23.9±0.9 mmHg/L/min, p=0.027), and SBP in daytime (144.1±1.4 vs. 151.6±2.1 mmHg, p=0.004) and nighttime (130.4±1.6 vs. 138.5±2.5 mmHg, p=0.008), and nighttime pulse pressure (PP, 47.2±0.9 vs. 51.7±1.4 mmHg, p=0.010) were significantly higher and cardiac output (5.4±0.1 vs. 4.8±0.2 L/min, p=0.043) and PP amplification (1.02±0.02 vs. 0.94±0.03, p=0.039) were significantly lower in male group 2. After multivariate analyses, SBP (β=0.225, p=0.010) and PP (β=0.179, p=0.022) at office, PWV (β=0.192, p=0.036), AIx (β=0.285, p=0.007), central SBP (β=0.351, p=0.001), DBP (β=0.246, p=0.012), and PP (β=0.317, p=0.006), SBP in daytime (β=0.219, p=0.004) and nighttime (β=0.232, p=0.013), and PP in 24-hour (β=0.216, p=0.005) and nighttime (β=0.172, p=0.024) were linearly associated with levels of TSH in hypertensive male.

Conclusion:
Despite euthryoid, arterial stiffness, central SBP, and SBP in ABPM are higher and cardiac output and PP amplification are lower in high normal TSH group of treatment-naïve hypertensive male, but not female. Arterial stiffness, SBP and PP at office, central BPs, and SBP and PP in ABPM are linearly associated with TSH levels among hypertensive male.
Abstract 217: Analysis Of Aspects Electrocardiographic, Vectorcardiographic And Echocardiographic Of Left Ventricular Hypertrophy In Resistant Hypertension
Authors: Dinamarco Nelson , Dinamarco Marileide , Moreno Heitor
Published: March 2022

Introduction:
Resistant hypertension (RHTN) defined as the failure to reach goal BP, with at least 3 drugs including a diuretic (JNC-VII). Affects 5-30% HTN, become high risk of developing target organ damage, like LVH rasing morbi-mortality (Framingham Study). Then is necessary a better treatment plan the early diagnosis. The electrocardiogram (ECG) and the vectorcardiogram (VCG), graphic methods (GM), alone, present, in spite of high specificity, have low sensitivity compared with echocardiography (ECHO).

Objective:
Analyze ECG/VCG for LVH diagnosed by ECHO in RHTN.

Methods:
Population: 120 RHTN. Excluded diabetic, valve heart, coronary, renal diseases and obese. Study Protocol: After 06 months, between 2008 and 2011, measures BP monthly (JNC-VII) receiving directions, as well as pill counts (Taylor Method) to assess compliance. Underwent ECHO (ASE) then GM. ECG&VCG methods: ECG Criterion: Sokolow-Lyon I(SL-I):SV1+RV5(6)≥3.5mV & II(SL-II):RavL≥1.1mV; Gübner(G):SDI+RDIII≥2.5mV;Cornell(C):RavL+SV3≥2/2.8mV(woman/man);Romhilt-Estes(RE):Point Score≥5points; Perugia(P): RE or C(modify) or Strain. VCG Criterion: Mag Max QRS on Planes horizontal(HP) & frontal(FP): vector≥2mV and right saggital(rSP): vector≥1.6 mV; Defl QRS HP & rSP: angle≥ 330° & ≥150°.

Statistical Analysis:
Sensibility(SENS), accuracy(ACCU) and specificity(SPEC) to analyse GM.

Results:
Criterion Sens (%) Accu (%) Spec (%)

SL-I 27 (16/60) 61 (73/120) 93 (56/60)

G 18 (11/60) 58 (69/120) 97 (58/60)

SL-II 18 (11/60) 58 (69/120) 97 (58/60)

C 25 (15/60) 59 (71/120) 93 (56/60)

RE 32 (19/60) 58 (77/120) 95 (58/60)

P 50 (30/60) 68 (81/120) 85 (51/60)

Mag Max QRS PH 28 (17/60) 61 (73/120) 93 (56/60)

Mag Max QRS PSr 12 (07/60) 52 (62/120) 92 (55/60)

Mag Max QRS PF 25 (15/60) 57 (68/120) 88 (53/60)

Defl QRS PH 12 (07/60) 52 (62/120) 92 (55/60)

Defl QRS PSr 13 (08/60) 50 (60/120) 87 (52/60)

COMBINED* 70 (42/60) 76 (91/120) 82 (49/60)

Conclusion:
ECG: P had the best SENS & ACCU, SL-II and G both showed the best SPEC. VCG: best criteria Mag Max QRS PH. Combined criteria* using best SENS and ACCU of each GM improved SENS & ACCU demonstrated SPEC without injury to LVH in RHTN detection. Can be worn with method in a cost effectiveness ratio and in places where access is difficult to ECHO to be performed.
Abstract 218: Role of Thioredoxin 1 in the Impaired Renal Sodium Excretion in D5F173L Transgenic Mice
Authors: Tan Xiaorong , Cai Jin , Ren Hongmei , Zhou Lin , Zeng Chunyu
Published: March 2022

Dopamine receptor plays an important role in hypertension by regulation of sodium excretion. Disruption of the D5 receptor results in hypertension with increased activity of the sympathetic nervous system. D5 receptor mutant F173L has been found to be related with hypertension. To investigate the role of F173L in hypertension, we established F173L transgenic mice, it resulted that, as compared with wild-type mice, the blood pressure was higher, basal levels of sodium excretion was lower, and fenoldopam, a D1-like receptor agonist, mediated natriuresis and diuresis were impaired in F173L mice. Our further study found ROS might be involved in the hypertension of F173L mice, because ROS levels were higher in F173L, treatment of F173L with tempol reduced the ROS, lower the blood pressure and normalized the sodium excretion in F173L mice. Screening studies found thioredoxin (Trx) 1 expression was lower, while Na+-K+-ATPase activity was higher in kidney from F173L mice and F173L transfected mouse proximal tubule epithelial cells (mRPT). Supplement with Trx1 could restore Na+-K+-ATPase activity in F173L transfected mRPT cells. Those results were confirmed in primary cultured mRPT cells from wild-type and D5F173L transgenic mice. We conclude that D5 mutant increases blood pressure, possibly via decreased D5 receptor mediated sodium excretion by decreased Trx1 expression in kidney.

Introduction:
D5 receptor mutant F173L has been found to be related with hypertension. To investigate the role of D5 receptor in the development of hypertension, we establish D5F173L transgenic mice. Our preliminary study found those mice develop hypertension with impaired sodium excretion. The previous studies reported that the hypertension of D5-/- mice was closely related with oxidative stress. To investigate the role of ROS in the impaired renal function in F173L mice, we selected some genes related oxidative stress and found the mRNA and protein expression of Trx1 were significantly decreased in the kidney of D5F173L transgenic mice.

Result:
We found that D5 mutant F173L increases blood pressure, via decreased Trx1 expression, decreased D5 receptor mediated renal sodium excretion, consequently leads to hypertension.
Abstract 220: Nephron Deficient Rats are Highly Susceptible to Hypertension Induced Kidney Injury
Authors: Wang Xuexiang , Johnson Ashley C , Guo Zibiao , Chen Fang , Solberg-Woods Leah , Garrett Michael R
Published: March 2022

It is well known that hypertension can promote decline in renal function, but there is a paucity of information on how hypertension influences single kidney individuals. The HSRA model exhibits a high incidence of congenital single kidney (50-75% offspring) that is characterized by low nephron number (~20% less in solitary kidney vs. control kidney), glomerular and kidney hypertrophy, elevated single nephron glomerular filtration rate (GFR), which culminates in kidney injury and decline in kidney function. We hypothesized that renal hemodynamic stressors, such as DOCA induced hypertension, would accelerate onset and progression of kidney injury in rats born with a solitary kidney (HSRA-S) compared to two-kidney littermates (HSRA-C) or rats that have undergone unilateral nephrectomy (HSRA-UNX). Time course evaluation of blood pressure, renal hemodynamics, and kidney injury was performed in 6 animal groups: (1-2) HSRA-S+DOCA or placebo; (3-4) HSRA-C+DOCA or placebo; and (5-6) HSRA-UNX+DOCA or placebo. At the conclusion of the study (5 weeks), HSRA-S+DOCA treated rats demonstrated a significant (ANOVA; p<0.05) rise in blood pressure (192±4 mmHg), proteinuria (205±31 mg/24hr) and decline in GFR (600+42 μl/min/gKW) relative to HSRA-UNX+DOCA (173±3 mmHg, 76±26 mg/24hr, 963+36 μl/min/gKW) and HSRA-C+DOCA (154±2 mmHg, 7±1 mg/24hr, 1484+121 μl/min/gKW). Animals in placebo groups showed no significant change in blood pressure, proteinuria or GFR from week 13-18 between all three groups. To elucidate the molecular mechanism of nephron deficiency in the HSRA model and better understand how elevated blood pressure promotes rapid decline in renal function, whole transcriptome analysis (RNA sequencing) of embryonic kidney isolated at key stages in nephrogenesis (E14.5, 15.5, and 16.5) was performed which identified a number of novel genes involved in nephrogenesis and kidney development. We are currently working to understand the functional role of these genes and pathways using an in vitro whole organ kidney culture system. In summary, nephron deficient rats are highly susceptible to hypertension induced kidney injury and several novel genes/pathways involved in nephrogenesis likely program the model to develop significant kidney disease with age.
Abstract 229: Genetic inhibition of Notch3 protects animals against Chronic Kidney Disease.
Authors: Keuylian Zela , El Machhour Fala , Dussaule Jean-Claude , Chatziantoniou Christos
Published: March 2022

In the present study, we show that expression of the Notch3 receptor in glomeruli is associated to the development of renal disease and that inhibition of this expression is accompanied by renoprotection.

In a model of rapidly progressive glomerulonephritis (nephrotoxic serum-induced renal disease), Notch3 expression was induced by several-fold in podocytes concurrently with disease progression. In contrast, mice with genetic ablation of Notch3 expression (KO) were protected as they showed less proteinuria, uremia and renal inflammation. In vitro studies suggested that podocytes expressing active Notch3 acquire a migratory and pro-inflammatory phenotype. To evaluate whether inhibiting Notch3 could be of therapeutic interest, we administered antisense oligodeoxynucleotides targeting Notch3 in wild type mice concomitantly to the disease induction, whereas scrambled sequences were used as controls. Both groups of mice developed renal disease, but mice injected with Notch3 antisense were protected compared to the scrambled group, as evidenced by the decreased values of plasma urea and proteinuria. In addition, the improvement of renal function was accompanied by fewer crescent formations and less deposits of fibrin within the glomeruli, and by decreased peritubular inflammation.

These results demonstrate that abnormal activation of Notch3 in the kidneys is involved in the progression of renal disease by promoting migratory and pro-inflammatory pathways and that blocking this activation preserves renal function and structure. Inhibiting Notch3 activation could be a novel, promising approach to treat chronic renal disease.
Abstract 237: Family History Of Premature Cardiovascular Events And Total Plaque Area
Authors: Perez Hernan A , Garcia Nestor H , Spence J. Dave , Armando Luis J
Published: March 2022

Although the positive family history for premature cardiovascular events has been considered a putative risk factor for decades, it has not been incorporated along with other established risk factors such as hyperlipidemia, hypertension, and cigarette smoking in the daily clinical practice when physicians evaluates the cardiovascular risk of a patient. One of the reason may be the presence of multiple risk factors coexisting during these evaluations.

The objective of this research was to investigate if patients with a family history of premature cardiovascular events have higher total plaque area than patients without it, in the absence of other classical risk factors.

Design and method:
Patients from the primary prevention database of Blossom DMO program were identified to have positive family history of premature cardiovascular events (FHPCVE). After apply excluding criteria (absence of any classical risk factor) 23 patients qualified. A control group was generated reaching same data in variables (blood pressure, age, body weight, LDL cholesterol), except carotid total plaque area which was reported at the end of groups formation, once both groups were formed. We used the same definition as the Framingham study (family history of <55 years in men and <65 years in women, first degree relatives) to consider a patient with FHPCVE.

Results:
The group with FHPCVE was similar to the control group. Age 59±2 sv 60±1 yo, blood pressure 126±2/75±1 mmHg vs 127±1/74±1 mmHg, body mass index, 23±1 vs 24±1 Kg/cm
2
and LDL cholesterol 116±15 vs 139±20 mg/dl. When total plaque area was determined by Doppler, patients with FHPCVE had a TPA of 49±8 mm
2
while control group 33±4 mm
2
(p<0.05). Framingham risk score were similar, while the post test for acute myocardial infarction was lower in control group (16±1% vs 20±3%, p<0.05).

Conclusions:
Our data indicates that patients with positive family history for premature cardiovascular events have larger total plaque area than patients without it, indicating enhanced risk to develop a cardiovascular event, even in the absence of other classical risk factor. Physicians should identify early and treat accordingly these patients to prevent arterial deterioration.
Abstract 248: Gender Dimorphism Of Endothelial Disorders In Hypertensives With Metabolic Risk Factors
Authors: Murashko Natalia , Bragina Anna , Bragina Galina , Podzolkov Valery
Published: March 2022

Objective:
To investigate endothelial function in men and women with essential hypertension (EH) in relation to metabolic risk factors.

Methods:
We examined 124 untreated patients (45 men and 79 women) with EH (mean age 50,3±5,6y and 52,3±7,6y, body mass index - 30,9±6,2 и 31,1±2,2 kg/m2 accordingly) (p>0,05) and 25 healthy volunteers (10 men and 15 women) with characteristics comparable. Plasma leptin levels were measured by radioimmunoassay, plasma levels of stable nitric oxide metabolites (NOx) - by spectrophotometry. Results were pro¬cessed with Statistica 6.0 software.

Results:
Prevalence of metabolic factors (visceral obesity, dyslipidemia and hyperglycemia) in men and women was comparable (p<0,05). Hypertensive males and females had higher NOx concentration than controls (49,5±19.1, 42,1±22.1 and 28,3±9,6 μmol/l, accordingly) (p<0,05) with predominance in males. In males with EH the highest NOx level was at grade 1(53,9±26,2), lower - at the grade 2(44,5±17) and the lowest - at the grade 3 EH 41,4±23,9 μmol/l (p<0,05). Females with EH didn’t have significant differences of NOx levels in accordance to EH grade: 42.6±2.2, 43.9±25.3, and 40.2±20.8μmol/l, accordingly (p>0,05). In males NOx levels correlated with EH grade (r=-0,36, p<0,05). NOx concentration did not differ between males and females with EH with or without dyslipidemia and hyperglycemia(p>0,05). Obese males with EH had higher NOx levels (39,5±14,9 μmol/l), than females (35,9±17,8 μmol/l) (p<0,05). Nonobese males and females with EH didn’t have significant differences of NOx concentration (55,3±17,9 и 53,8±20,1 μmol/l, accordingly) (p>0,05). In female NOx levels correlated with the presence of obesity (r=-0,48, p<0,05). Males with EH and high leptin levels had higher NOx concentration (31,6±13,1 μmol/l), than females (25,4±10,1 μmol/l) (p<0,05). NOx levels did not differ between males and females with EH and normal leptin levels (53,1±11,3 и 51,8±10,5 μmol/l, accordingly) (p>0,05).

Conclusion:
Our results demonstrate gender dimorphism of cardiovascular risk factors impact in endothelial dysfunction. The presence and grade of EH are more significant for male hypertensives while obesity and hyperleptinemia - for female hypertensives.
Abstract 252: Renal Rescue of D2R Function in Mice Using an AAV9 Vector Reverses the Renal Injury and High Blood Pressure Induced by D2R Silencing in the Kidney
Authors: Konkalmatt Prasad R , Asico Laureano D , Yang Yu , Jose Pedro A , Armando Ines
Published: March 2022

Lack or downregulation of dopamine D2 receptor (D2R) function in mice increases the vulnerability to renal inflammation. Recent studies from our laboratory showed that long-term (28 days) selective down regulation of D2R in one kidney in mice with two intact kidneys increases systolic blood pressure and the expression of inflammatory and injury markers. We hypothesized that rescuing D2R function by re-expressing the receptor in the kidney of mice with D2R silencing will reduce renal inflammation and injury, and normalize blood pressure. To test our hypothesis we silenced D2R expression by the renal subcapsular infusion of D2RsiRNA and then we rescued the silenced D2R expression using retrograde ureteral infusion of AAV9 vector carrying wild-type D2R (D2RAAV). Mice were treated with non-silencing siRNA (NS siRNA) or D2RsiRNA (3 μg/day, n=6 per group) using 28 day osmotic minipumps. Fourteen days following the initiation of siRNA treatment, each group was treated with control AAV (CAAV) or D2RAAV (n=3 per group, 1e+11 viral genome particles per mouse). Fourteen days following AAV administration, blood pressure was recorded and organs were collected. D2R expression was decreased in D2RsiRNA-treated kidneys compared with NS siRNA-treated kidneys (immunoblotting: 54 ± 0.8 vs 100± 22 %; P<0.05). D2RAAV treatment increased D2R expression (7.5 -11 fold; P<0.01) in both D2R and NS siRNA-treated mice. Mice treated with D2RsiRNA+CAAV had increased systolic blood pressure (121±3 mmHg; P<0.05) in comparison with mice treated with D2RsiRNA+D2RAAV (100±6 mm Hg) (rescued mice), NS siRNA +CAAV (101±4 mm Hg), or NS siRNA +D2RAAV (101±1 mm Hg). The expression of TGF-β was higher in the mice treated with D2RsiRNA+CAAV than in D2R rescued mice (D2RsiRNA+D2RAAV) (100±22 vs 54± 11%; P<0.05). The number of cells positive for the marker of proliferation ki-67 was higher in kidneys of D2RsiRNA+CAAV mice than in D2RsiRNA+D2RAAV (32± 5 vs 15±2 per field; P<0.01). In conclusion, these results demonstrate that increased blood pressure and renal injury due to D2R silencing could be rescued by over expression of D2R using AAV9 vector in the kidney. Furthermore, these data confirms the protective effects of renal D2R and may provide the basis for designing novel therapies for kidney disease.
Abstract 253: Norepinephrine Suppresses T-lymphocyte Growth and Cytokine Production via Superoxide During Hypertension
Authors: Case Adam J , Zimmerman Matthew C
Published: March 2022

Numerous studies have shown that pro-inflammatory cytokines produced by T-lymphocytes exacerbate hypertension. However, it is not clearly understood how endogenous pro-hypertensive stimuli such as norepinephrine (NE) directly act upon T-lymphocytes to modulate the responses of these inflammatory cells during hypertension. Herein, we hypothesized that increased systemic levels of NE directly activate T-lymphocytes during hypertension. To test this hypothesis, we examined the status of T-lymphocyte activation in the spleens of NE-infused hypertensive mice. Contrary to our hypothesis, while subcutaneous infusion of NE (3.8 μg/kg/min) significantly increased mean arterial pressure by approximately 18 ± 2 mmHg for 14-days (p<0.05 vs. saline infused), splenic T-lymphocytes demonstrated no signs of increased activation. More specifically, splenic T-lymphocytes from NE-infused mice showed an approximate 20% ± 5% (p<0.05) decrease in proliferation accompanied by a 50% ± 17% (p<0.05) and 85% ± 6% (p<0.05) reduction in interferon gamma (INFγ) and tumor necrosis factor alpha (TNFα) production respectively as compared to T-lymphocytes from saline-infused mice. Additionally, NE directly inhibited naïve T-lymphocyte proliferation and cytokine production
ex vivo
in a dose dependent manner. Furthermore, while NE did not demonstrate any pro-apoptotic effects on T-lymphocytes, a 21% ± 2% (p<0.05 vs. saline) increase in G1 arrest was observed in NE-treated T-lymphocytes, and this was accompanied by a 60% ± 4% (p<0.05 vs. saline) decrease in pro-growth cyclin D3, E1, and E2 mRNA expression. Interestingly, NE caused a 71% ± 17% (p<0.05 vs. saline) increase in cellular superoxide (O
2
•-
) production as evidenced by dihydroethidium (DHE) oxidation, which was shown to be partially-causal to the inhibitory effects of NE as the addition of Tempol, a O
2
•-
scavenger, to the drinking water of NE-infused mice was able to moderately restore T-lymphocyte growth and pro-inflammatory cytokine production. Taken together with previous studies, our data indicates that direct NE stimulation of naïve T-lymphocytes inhibits their proliferation and cytokine production in hypertension.
Abstract 256: AMP-activated Protein Kinase Activator AICAR Attenuates TNFα-induced Inflammation In Murine Adipocytes.
Authors: White Anna , Nguyen Dinh Cat Aurelie , Jenkins Carol , Mancini Sarah J , Montezano Augusto C , Salt Ian P , Touyz Rhian M
Published: March 2022

Adipose tissue, a metabolically active tissue and important regulator of vascular function, contributes to obesity-related cardiovascular diseases such as hypertension. Molecular mechanisms are still elusive, but inflammation and activation of a local renin-angiotensin-aldosterone system (RAAS) seem to play an important role in adipocyte dysfunction leading to vascular damage. AMP-activated protein kinase (AMPK), a key regulator of cell metabolism, may play an important role in adipose tissue biology/inflammation. In this study, we evaluated whether AMPK activation regulates TNFα-induced inflammation and RAAS status in adipocytes. Murine adipocytes (3T3-L1) were used and treated with 10ng/ml TNFα in the absence/presence of AICAR (1mM - AMPK activator). Pro-inflammatory markers were assessed by qPCR or ELISA and protein expression by immunoblotting. TNFα stimulation increased IL-6 (113.4 fold) and MCP-1 (449.6 fold) mRNA levels, as well as IL-6 release (17.67 fold) into culture medium (p<0.05, vs vehicle); an effect abrogated by AICAR (75% reduction MCP-1 mRNA, 60% reduction IL-6 mRNA and 30% reduction IL-6 release). Adiponectin mRNA levels were decreased by TNFα (60%, p<0.05 vs vehicle), yet preincubation with AICAR had no effect. The effects of TNFα were also evaluated on components of the RAAS. TNFα induced a decrease in angiotensinogen (60%), mineralocorticoid receptor (70%) and AT2R (90%) mRNA levels in adipocytes (p<0.05), which was unaffected by AICAR. Activation of AMPK also suppressed TNFα-stimulated phosphorylation of MAP kinases and NFκB assessed by immunoblotting. In conclusion, AMPK activation seems to play a role in the control of inflammation and RAAS in adipocytes. Our data highlights the importance of AMPK as an important anti-inflammatory signaling protein in adipose tissue and may be an interesting target for treatment of obesity-related vascular injury leading to hypertension. This is especially significant given that the commonly used drug metformin is an AMPK inhibitor.
Abstract 257: miR-217 Mediates the Protective Effects of the Dopamine D2 Receptor on Renal Inflammation and Fibrosis.
Authors: Han Fei , Jiang Xiaoliang , Konkalmatt Prasad , Yang Yu , Gildea John , Jones John E , Felder Robin A , Jose Pedro A , Armando Ines
Published: March 2022

Some common single nucleotide polymorphisms (SNPs; rs 6276, 6277, and 1800497) in the human dopamine D2 receptor (DRD2) gene are associated with decreased receptor expression and function, as well as high blood pressure. We have reported that human renal proximal tubule cells (hRPTCs) from subjects carrying D2R SNPs had decreased D2R expression and function and increased expression of the pro-fibrotic factor TGFβ1 and its signaling targets Smad3 and Snail1. These cells also showed induction of epithelial mesenchymal transition and production of extracellular matrix proteins fibronectin-1 (FN-1), Collagen 1a (Col 1a), and vimentin. To determine the mechanisms underlying the effects of the D2R SNPs, we studied the expression of several microRNAs (miRs) that are associated with the regulation of chemokines, anti-fibrosis, extracellular matrix remodeling, and cell adhesion through post-transcriptional repression of their target mRNAs. The expression of 4 miRs (miR-217, miR-224, miR-335, miR-1265) was downregulated (-10.3 to -6.0 fold) and that of one miRNA (miR-1290; 5.3 fold) was upregulated in hRPTCs with D2R SNPs. The expression of miR-217 and miR-224 is regulated by the D2R; the expression is decreased by D2R downregulation in cells not carrying D2R SNPs (WT) and normalized by treating cells carrying D2R SNPs with a Drd2 plasmid. Treatment of WT cells with a miR-217 mimic significantly decreased the mRNA expression of TGFβ1 (43±2%), MMP3 (65±4%), and FN-1 (30±4%) while treatment with a miR-217 inhibitor increased TGFβ1 (94± 9%), MMP3 (96±5%), and FN-1 (212±30%). miR-217 mimic treatment of D2R SNPs cells also significantly decreased the expression of TGFβ1, MMP3, and FN-1. However, treatment of these cells with a miR-217 inhibitor had non-significant effects probably because miR-217 level was already reduced. In contrast treatment of either WT or D2R SNPs cells with miR-224 mimic or inhibitor had no significant effects on the expression of TGFβ1, MMP3, or FN-1. These results indicate that the protective effects of D2R on inflammation and fibrosis are at least, in part, mediated by regulation of miR-217 expression. Dysregulation of miR-217 expression may be responsible for the profibrotic phenotype of hRPTCs in individuals with these D2R SNPs.
Abstract 258: The Compensatory Effect Of An Increased Body Of Myeloid-derived Suppressor Cells In Hypertension
Authors: Shen Xiao , Shah Kandarp , Shi Peng , Giani Jorge , Li You , Bernstein Kenneth
Published: March 2022

The immune system plays a critical role in hypertension progression. It has been showed that the lack of T cells prevented the development of hypertension. However, the role of myeloid cells in this process is elusive. We now show that there is a 2 fold increase of CD11b+ Gr1+ myeloid cells in murine hypertension models induced by angiotensin II, L-NAME or high salt diet. These increased myeloid cells are immunosuppressive in that they could suppress T cell proliferation. The production of reactive oxygen species mediates their immunosuppressive functions. When hypertensive mice were depleted of these myeloid cells, using either anti-Gr1 antibody or gemcitabine, there was a 15 mmHg further rise in blood pressure and aggravation of T cell activation as manifested by increased production of IFN-γ, TNFα and IL-17 in both spleen and kidney. In contrast, adoptive transfer of myeloid-derived suppressor cells could be used as both treatment reagent to alleviate established hypertension and preventive reagent from hypertensive insult. These data unveil the role of a distinctive myeloid population in hypertension, which may be a potential target for hypertension management.
Abstract 263: Toll-like Receptor 4 Involvement in Normal Bladder Smooth Muscle Contractile Function
Authors: Szasz Theodora , Nunes Kenia P , Carrillo-Sepulveda Maria Alicia , Webb R Clinton
Published: March 2022

Overactive bladder (OAB) and bladder dysfunction are commonly associated with aging and metabolic disorders, and both hypertension and the use of anti-hypertensive medication have been correlated with OAB. Recent evidence demonstrates the involvement of the innate immune system in chronic disease conditions via Toll-like receptors (TLRs) activation by damage associated molecular patterns (DAMPs) released from injured cells. TLR4 is expressed in both urothelium and detrusor layers of the bladder and we have previously shown that TLR4 activation contributes to diabetic bladder dysfunction. Here we hypothesized that TLR4 participates in the physiological function of bladder smooth muscle cells. We used wild type (WT) and TLR4-/- (TLR4KO) adult male mice and evaluated the bladder smooth muscle contractile function. We observed that bladder weight was increased in TLR4KO compared to WT mice (% body weight: WT=0.98±0.03, TLR4KO=1.20±0.07). Contraction of urothelium-denuded bladder strips to carbacholine was increased in TLR4KO compared to WT mice (pD2: WT=5.74±0.11, TLR4KO=6.22±0.08). TLR4KO bladder also displayed a higher electrical field stimulation (EFS)-induced maximal contractile response compared to WT, both in the presence and absence of urothelium (mN force at 32 Hz: WT-uro=22.96±2.33, TLR4KO-uro=30.98±6.45, WT+uro=33.05±4.57, TLR4KO+uro=45.30±3.61). Cyclooxygenase 1 (COX1) expression was increased in bladder smooth muscle from TLR4KO compared to WT. COX inhibition by indomethacin reverted the augmented contractile response of TLR4KO bladder strips to carbacholine. No differences in basal reactive oxygen species levels were observed between bladder smooth muscle cells isolated from WT and TLR4KO. These data demonstrate that genetic ablation of TLR4 leads to bladder hypercontractility via COX activation and suggest that TLR4 contributes to the maintenance of physiological smooth muscle function of the bladder.
Abstract 270: Successful Development and Implementation of an Algorithm to Identify Resistant Hypertension in the Electronic Medical Record
Authors: Shuey Megan M , Denny Joshua C , Edwards Todd L , Brown Nancy J
Published: March 2022

Resistant hypertension (RH) affects 8.4 to 17.4% of the hypertensive population, and is associated with an elevated risk of end-organ damage. To identify genetic or environmental factors associated with RH, we developed an algorithm to phenotype patients using electronic medical records (EMRs).

We modified a previously validated algorithm for detecting patients with RH from 158,128 adults in BioVU, a DNA databank incorporating clinical data from EMRs. We defined patients as having RH if they had a SBP>140 mmHg or a DBP>90 mmHg for at least one month while receiving at least three anti-hypertensive agents including 25 mg HCTZ or equivalent doses of another thiazide diuretic or amlodipine 5 or 10 mg, or equivalent doses of a similar calcium channel blocker. Patients could be controlled after addition of a fourth drug. Patients were defined as having controlled hypertension if they carried a diagnosis of hypertension, were taking at least one and no more than two anti-hypertensive medications, and had a SBP<135 and a DBP<90 mmHg on repeated measurement. Drug exposures were identified from electronic-prescribing tools and MedEx.

To validate the algorithm, we conducted an independent review of 150 EMRs from patients classified as having resistant or controlled hypertension. The accuracy of the algorithm was 94.7%. Among 55,477 patients with hypertension in BioVU, 14.2% met the criteria for RH. The table shows the characteristics of patients with resistant and controlled hypertension.

This study demonstrates the successful use of a phenotyping algorithm to identify subjects with RH from EMR data and provides important characteristics of patients with RH in a real-world clinical setting.
Abstract 279: Vascular Aging In Aldosterone Associated Hypertension: Role Of NADPH Oxidase 1
Authors: Harvey Adam , Montezano Augusto , Graham Delyth , He Ying , Ceravolo Graziela , Yabe-Nishimura Chihiro , Griendling Kathy , Touyz Rhian
Published: March 2022

In hypertension, vascular aging is accelerated leading to heart failure, stroke and renal dysfunction. Cellular and molecular mechanisms of age-associated vascular changes are unclear, but enhanced ROS generation by NADPH oxidases (Nox) and aldosterone (aldo) have been suggested. Nox1 is of particular interest due to increased levels in hypertension. Here, we postulated that aldo plays an important role in vascular aging through Nox1-dependent mechanisms. We examined signalling molecules associated with aging in arteries from 2 experimental models: stroke-prone spontaneously hypertensive rats (SHRSP) rats and Nox1 knockout mice infused with aldo (300 ug/Kg/day). Gene expression was assessed by qPCR and protein levels by immunoblotting. Aldo was measured by ELISA. In SHRSP rats, aging-associated mRNA inflammatory markers, such as RANTES (5- fold), MCP-1 (6-fold) and IL-6 (2-fold); as well as aldo levels (6-fold) and H2AX (marker of aging-associated DNA damage - 1.5-fold) were increased compared with control rats, p<0.05. In mice treated with aldo, JNK (pro-inflammatory - 69%) and p66SHC (pro-senescence - 92%) activation were increased in mesenteric arteries (p<0.05); an effect blunted in vessels from Nox1 KO mice. In parallel studies, vascular smooth muscle cells (VSMCs) from adult and aged control mice, as well as, from adult Nox1 transgenic mice (Nox1 overexpression in VSMCs) were extracted and stimulated with H2O2 and aldo. Basal levels of p66SHC (39%) and OGG-1 (48%), markers of senescence, were increased in VSMCs from aged animals, p<0.05. H2O2 (64%) and aldo (84%) increased p66SHC activation in VSMCs from adult mice to similar levels observed in cells from aged animals (p<0.05). However, aldo effects on p66SHC activation were exacerbated in VSMCs from adult Nox1 transgenic mice. In conclusion, aldosterone may play an important role in the aging-like phenotype in vascular injury associated with hypertension. Such processes may involve Nox1 and redox-sensitive p66Shc signalling.