bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 245: Role Of Melanocortin-4-receptor In The Blood Pressure Regulation In Female Hyperandrogenemic Rats, A Model Of Polycystic Ovary Syndrome
Authors: Maranon Rodrigo O , Lima Roberta , do Carmo Jussara M , Da Silva Alexander , Hall John E , Reckelhoff Jane F
Published: July 2021

Women with polycystic ovary syndrome (PCOS) are characterized by hyperandrogenemia, hirsutism, infertility, and enlarged cystic ovaries. Obesity and hypertension are also frequently found in women with PCOS, although the mechanisms responsible for the elevated blood pressure (BP) are unclear. Thus we tested the hypothesis that the melanocortin-4 receptor (MC4R), known to contribute to obesity hypertension in males, contributes to the elevated BP in hyperandrogenemic female rats, a model of PCOS. Female Sprague Dawley rats (4 wks) were implanted with dihydrotestosterone (DHT; 7.5mg/90 days sc) or placebo pellets (PL) (n=5/grp) and aged to 12 wks. Body weight and food intake (whether rats were pair fed or had ad libitum access) were measured daily. Two wks following implantation of radiotelemetry transmitters and intracerebroventricular cannulae, baseline mean arterial pressure (MAP) was measured for 5 days; then rats received MC3/4R antagonist, SHU-9119 (SHU; 1 nmol/h ICV) or vehicle for 7 days and MAP was recorded. DHT-treated rats had higher body weight and MAP than PL rats (BW: PL: 266.0±8.7; DHT: 348.5±10.4 g, p<0.01; MAP: PL rats: 102±5; DHT: 114±5 mmHg, p<0.05). SHU significantly increased food intake and body weights in both placebo (PL) and DHT-treated rats fed ad libitum (PL: 379.2±28.5; DHT: 451/3±7.3 g, p<0.01 DHT vs PL; 0.01 SHU vs control), but had no effect on MAP compared to controls (PL: 104±5; HAF rats: 114±5 mmHg; p<0.05, HAF vs PL; p=NS, SHU vs controls). However, in other rats, when pair fed with little increase in body weight (PL: 253.7±2.0, SHU: 261.0±0.6, p<0.05; DHT: 306.7±2.6, SHU: 316.7±1.5 g, p<0.05), SHU decreased MAP in DHT treated rats but not placebo controls (PL rats: 102±1, SHU: 103±2 mmHg; p=NS; DHT rats: 110±1 vs. SHU: 97±1 mmHg; p<0.001). Thus MC4R antagonist reduces MAP in DHT-treated rats only when food intake and body weight are controlled. These data suggest that activation of MC4R contributes to elevated BP in our model of PCOS and may also contribute to the elevated BP in women with PCOS. Supported by NIH R01HL66072, P01HL05971 and AHA 14POST18640015.
Abstract 259: C-reactive Protein Ultrasensitive (hscrp) In Resistant Hypertensive Patients (rhtn) With Left Ventricular Hypertrophy (lvh)
Authors: Dinamarco Nelson , Dinamarco Marileide , Moreno Heitor
Published: July 2021

Introduction:
Left ventricular hypertrophy (LVH) is an important cardiocerebrovascular risk factor and due to this fact, when present , in hypertensive patients should plan a more elaborate therapeutic treatment. The hsCRP is systemic inflammatory marker associated with increased risk of myocardial infarction and vascular diseases.

Objective:
Identify hsCRP as a strong biomarker with correlation with LVH in resistant hypertensive patients (RHTN).

Methodology:
A total of 180 patients: 60 RHTN and 60 RHTN with LVH with echocardiographic diagnosis (LVH-RHTN), compared to 60 normal subjects (control), with equal distribution of gender and age. The hs-CPR was measured using nephelometry method.

Results:
In LVH-RHTN hsCRP was 3.96 ± 1.11 mg/L and was significantly higher when compared with RHTN, which was 2.20 ± 0.53 mg/L, and with hsCRP 0.56 ± 0.23 in CON (p <0.001). There is a strong correlation between elevated hsCRP and LVH in patients with resistant hypertension, as shown by the Pearson coefficient (r = 0.74).

Conclusion:
hsCRP can be considered a strong predictor biomarker in presence of LVH in RHTN.
Abstract 278: The Glucocorticoid Receptor Antagonist RU486 Ameliorates Cold Stress-induced Exacerbation of Cardiac and Adipose Tissue Pathology and Metabolic Disorders in a Rat Model of Metabolic Syndrome
Authors: Nagasawa Kai , Matsuura Natsumi , Minagawa Yuji , Ito Shogo , Sano Yusuke , Hattori Takuya , Yamada Yuichiro , Watanabe Shogo , Murohara Toyoaki , Nagata Kohzo
Published: July 2021

Introduction:
Chronic stress, when combined with hyperphagia, can affect adiposity and metabolism. However, few studies have reported the effects of cold stress on cardiovascular and metabolic disorders in metabolic syndrome (MetS). We investigated the effects of chronic cold stress and glucocorticoid receptor (GR) blockade on cardiac and adipose tissue pathology and gene expression and on glucose and lipid metabolism in a rat model of MetS.

Methods and Results:
We used DahlS.Z-Leprfa/Leprfa (DS/obese) rats which are derived from a cross between Dahl salt-sensitive and Zucker rats and represent a new animal model of MetS. DS/obese rats were exposed to cold stress (ice-cold water, 1 cm depth, 2 h/day) for 4 weeks beginning at 9 weeks of age with or without the GR antagonist RU486 (2 mg/kg/day, sc). Age-matched homozygous lean (DahlS.Z-Lepr+/Lepr+, or DS/lean) littermates of DS/obese rats served as controls. Chronic cold stress exacerbated hypertension as well as left ventricular (LV) hypertrophy, fibrosis and diastolic dysfunction, in a manner sensitive to RU486. Cold stress and RU486 did not affect body weight or visceral and subcutaneous fat mass. In contrast, cold stress further increased superoxide production and NADPH oxidase activity in the heart as well as macrophage infiltration and the expression of proinflammatory genes in LV and visceral fat tissue. RU486 treatment inhibited these changes in gene expression, as well as cardiac oxidative stress and inflammation and adipose tissue inflammation. Cold stress further up-regulated cardiac renin-angiotensin-aldosterone system gene expression as well as the expression of GR and 11β-hydroxysteroid dehydrogenase type 1 genes in LV and visceral adipose tissue, and all of these effects were attenuated by RU486. In addition, RU486 ameliorated the stress-induced deterioration of dyslipidemia (elevations in low-density lipoprotein cholesterol, triglycerides, and free fatty acid) as well as that of glucose intolerance and insulin resistance.

Conclusions:
The present results indicate that GRs may be involved in cold stress-induced exacerbation of cardiac and adipose tissue pathology as well as that of glucose and lipid metabolism in a rat model of MetS.
Abstract 336: Acute Increase In O-GlcNAcylation Reduces The Release Of Cytokines And Attenuates Hypotension In Mice With LPS-induced Sepsis.
Authors: Olivon Vania C , Ferreira Raphael G , Mestriner Fabíola L , Alves-Filho José C , Cunha Fernando Q , Tostes Rita C
Published: July 2021

The attachment of β-O-linked N-Acetylglucosamine (O-GlcNAc) in nuclear and cytoplasmic proteins, known as O-GlcNAcylation, is a common post-translational modification controlled by two enzymes: O-GlcNAc transferase (OGT) and β-N-acetylglucosaminidase (OGA). Acute increases in O-GlcNAc reduce migration of inflammatory cells and the release of pro-inflammatory mediators, important events in sepsis caused by bacterial infection or multiple non-infectious causes. We postulated that acute increases of O-GlcNAc reduce sepsis-associated mortality, release of inflammatory mediators and changes in blood pressure and vascular reactivity. C57BL/6 mice received lipopolysaccharide (LPS) injections to produce mild (LPS M, 10mg/Kg, i.p.) or severe (LPS S, 20mg/Kg, i.p.) sepsis. LPS-treated and naive (N) mice received glucosamine (GlcN), 300mg/Kg, i.v.) or vehicle (30 min before the LPS administration) and were euthanized 6 h later. GlcN treatment augmented O-GlcNAc levels and increased survival in mice with LPS-induced sepsis (LPS M=20%*; LPS S=50%*). GlcN treatment reduced serum levels (pg/mL) of IL-1β [N and N+GlcN = not detected (nd); LPS M = 286.3±14.5*, LPS M+GlcN = 212.9±3.1*#; LPS S= 343.9±29.1*, LPS S+GlcN = 128.4±11*#], IL-6 (N and N+GlcN = nd; LPS M = 448.0±11.5*, LPS M+GlcN = 241.2±11.6*#; LPS S = 508.6±21.7*, LPS S+GlcN = 451.6±8.9*) and TNF-α ((N and N+GlcN = nd; LPS M = 311.3±15.7*, LPS M+GlcN = 76.5±5.5*#; LPS S = 354.2±32.1*, LPS S+GlcN = 136.2±10.2*#) and vascular mRNA expression (2-ΔΔCT) of these cytokines: IL-1β (N and N+GlcN = nd; LPS M = 20.6±1.2*, LPS M+GlcN = 14.0±1.4*#; LPS S = 21.46±1.5*, LPS S+GlcN = 15.87±0.9*#), TNF-α [N and N+GlcN = nd; LPS M = 0.07±0.004*, LPS M+GlcN = 0.02±0.004*#; LPS S = 0.13±0.01*, LPS S+GlcN = 0.02±0.006*#. GlcN treatment attenuated LPS-induced decrease in blood pressure [(mmHg), N = 108±9.0, N+GlcN = 112±10.0; LPS M = 72±6.0*, LPS M+GlcN = 92±8.5*#; LPS S = 59±4.5*, LPS S+GlcN = 88±7.5*# mmHg). No changes in vascular reactivity (thoracic aorta) to phenylephrine or acetylcholine were detected 6 h after LPS administration; (* p<0.05 vs. N; # p<0.05 vs. LPS). In conclusion, O-GlcNAc reduces sepsis-associated inflammatory and cardiovascular events, making this pathway a potential target for therapeutic intervention.
Abstract 348: Inhibition Of Phosphodiesterase 5 Attenuates Angiotensin II-dependent Hypertension And Renal Vascular Dysfunction In C57bl/6 Mice But Not In Enos-Ko Mice
Authors: Thieme Manuel , Sivritas Sema , Potthoff Sebastian A , Mergia Evanthia , Rump Lars C , Stegbauer Johannes
Published: July 2021

The kidney plays an outstanding role in the blood pressure (BP) regulation. The renal vasoconstrictor response to angiotensin (Ang) II is balanced by the NO/cGMP-signalling cascade. Ang II causes hypertension and vascular dysfunction by reducing cGMP sensitivity. Ang II is able to increase cGMP degradation by activating phosphodiesterase (PDE)1 and PDE5. The aim of the present study was to identify the predominant PDE subunit regulating renal blood flow (RBF) and vascular tone during hypertension. Therefore, we tested in vivo effects of acute PDE1 (vinpocetine) and PDE5 (sildenafil) inhibition at baseline and during acute Ang II infusion (200ng/kg/min). Furthermore, we examined the impact of PDE-inhibition on Ang II dependent hypertension (500ng/kg/min; 14 days) and on renal vascular function in the isolated perfused kidney.

Acute vinpocetine administration (0.8-800μg/kg BW) showed almost no effect on systemic BP and RBF at baseline and during acute Ang II infusion. In contrast, sildenafil (0.8-800μg/kg BW) significantly decreased BP under baseline conditions. During acute Ang II infusion, BP reduction and RBF increase induced by sildenafil was even more pronounced suggesting a pivotal role of the PDE5 in the regulation of renal vascular tone. Based on these results, we tested whether inhibition of the PDE5 protects from hypertension and vascular dysfunction. Indeed, chronic sildenafil treatment significantly attenuated Ang II dependent hypertension in C57BL/6 (vehicle vs. sil: 156±4 vs. 139±7; p<0.05). Moreover, Sildenafil treatment significantly improved NO-dependent vasorelaxation in kidneys of Ang II- treated C57BL/6. To confirm that PDE5 is activated by an increased NO/cGMP signaling, we used eNOS-KO mice, a model known for decreased NO dependent cGMP generation. In eNOS-KO mice, sildenafil failed to reduce Ang II dependent hypertension (172,4 ± 4,3 mmHg vs. 166,1 ± 3,8 mmHg, p=0,2753) and did not improve vascular dysfunction in Ang II treated kidneys.

In summary, the PDE5 is the predominant PDE regulating RBF. Inhibition of PDE5 by sildenafil ameliorates chronic Ang II dependent hypertension and improves vascular dysfunction. This study reveals new evidence for the pivotal role of PDE5 in the pathogenesis of AngII-induced hypertension.
Abstract 401: Immuno-phenotypic Characterization of Lateralized Primary Aldosteronism with New Monoclonal Antibodies against the Human Aldosterone Synthase (CYP11B2) and 11- beta hydroxylase (CYP11B1)
Authors: Gioco Francesca , Gomez-Sanchez Celso , Fassina Ambrogio , Qi Xin , Velarde-Miranda Carolina , Rossitto Giacomo , Gomez-Sanchez Elise , Rossi Gian Paolo
Published: July 2021

Background:
Primary Aldosteronism (PA) is commonly due to an Aldosterone-Producing Adenoma (APA), but until now the lack of a specific antibody for human aldosterone synthase (hCYP11B2) has prevented the identification of APA in the excised adrenal gland.

Objectives:
To investigate the usefulness of a novel monoclonal antibody for hCYP11B2 for the immuno-phenotypic characterization of a large series of consecutive adrenal glands from APA patients diagnosed by the “four corners criteria” fully characterized for KCNJ5 mutations.

Methods:
To generate the antibodies, spleen cells from mice immunized with peptides against hCYP11B2 were fused to mouse myeloma cells. CYP11B2 and CYP11B1 were detected with immunohistochemistry in the adrenals from 49 consecutive APA patients. Based on results of a pilot study the following classification was developed: Pattern1: adenoma with uniform CYP11B2 staining + few areas of CYP11B1 staining; Pattern2: adenoma with diffuse CYP11B2 staining + few areas of CYP11B1 staining + some cells co-expressing CYP11B2/CYP11B1; Pattern3: adenoma expressing CYP11B1 + clusters of cells CYP11B2; Pattern4: no adenoma + multiple clusters of sub-capsular CYP11B2 cells; Pattern5: multi-nodular adrenal cortex + clusters of sub-capsular CYP11B2 cells.

Results:
KCNJ5 mutations were found in 38% of the cases (19/49). With CYP11B2/CYP11B1 immunostaining the following results were found: pattern 1 16.3% (8/49); pattern2 32.7% (15/49); pattern 3 16.3% (7/49); pattern 4 22.4% (10/49); pattern 5 12.2% (5/49).

Conclusions:
The highly variable steroidogenic enzymes (CYP11B2, CYP11B1) expression in a series of consecutive APA show the existence of at least 5 different patterns, thus challenging the classical notion that PA entails only APA or bilateral hyperplasia.
Abstract 455: M2 Macrophages Contribute to Vessel Fibrosis and Stiffening During Angiotensin II-Dependent Hypertension in Mice
Authors: Drummond Grant R , Moore Jeffrey P , Tuck Kellie L , Sakkal Samy , Krishnan Shalini M , Diep Henry , Kemp-Harper Barbara K , Budzyn Klaudia , Ricardo Sharon D , Guzik Tomasz J , Samuel Chrishan S , Sobey Christopher G
Published: July 2021

Arterial fibrosis and stiffening are hallmarks of hypertension and contribute to elevated systolic blood pressure (BP), heart failure and end organ damage. Hypertension is also associated with accumulation of macrophages in the vascular wall; however, the phenotype of these cells (M1/M2) and their contribution to vascular fibrosis/stiffening remain to be established. Chronic angiotensin II-infusion in mice (0.7 mg/kg/d, 28 d) caused a marked increase in systolic BP (169±6 mmHg vs 118±3 mmHg in saline-infused mice). Flow cytometric analysis of aortic cell suspensions revealed that angiotensin II infusion also caused a 2-fold increase in F4/80
+
macrophage numbers. Moreover, these cells were positive for the M2 marker, CD206. Quantitative PCR confirmed that aortic expression of CD206 and other M2 markers (arginase-1, Fc receptor-like S, scavenger receptor) were elevated in angiotensin II-infused mice, whereas levels of M1 markers (inducible nitric oxide synthase, CXCL2, tumour necrosis factor) were unchanged. Twice-weekly treatment of angiotensin II-infused mice with clodronate-encapsulated liposomes reduced numbers of circulating monocytes (i.e. macrophage precursors) by ~70%, whereas saline-containing liposomes had no effect. Clodronate treatment prevented M2 macrophage accumulation in the aorta, and also in the kidneys, and importantly reduced collagen deposition in these tissues. Clodronate treatment also improved arterial compliance and lowered BP by ~20 mmHg in angiotensin II-treated mice. In conclusion, M2 macrophages contribute to tissue fibrosis and aortic stiffening during angiotensin II-induced hypertension in mice. Thus, strategies that limit macrophage accumulation in the vessel wall and/or prevent their polarization towards an M2 phenotype may hold promise as future therapies for hypertension.
Abstract 456: Role of Signal Transducer and Activator of Transcription protein (STAT)-1 in the Angiotensin II-induced Cardiovascular Damage
Authors: Chmielewski Stefan , Aoqui Cristiane , Bluijssen Hans , Baumann Marcus
Published: July 2021

Background:
Inflammation participates importantly in hypertensive organ damage. Angiotensin II (Ang II) plays a crucial role in hypertension and induces inflammation. An essential mediator of inflammation is the transcription factor STAT1 which is activated by interferon but also by Ang II. We hypothesized that activation of STAT1 during Ang II infusion upregulates chemokines, enhances chemotaxis and consequently results in heart fibrosis and vessel dysfunction independent on blood pressure and hypertrophy

Methods:
C57BL/6, C57BL/6 receiving candesartan (5mg/kg) and STAT1-/- mice received infusion of Ang II 1.5 μg/g/day or placebo for 4 weeks (n=9/group). Blood pressure was measured using tail cuff pletysmography. Expression of chemokines Cxcl10, Cxcl9 and MCP-1 as well as Nos2 were investigated. Small mesenteric arteries (SMA) were mounted in a wire myograph to assess their function. Cardiac hypertrophy and inflammation (CD45 staining) and fibrosis (hydroxyproline assay) were determined.

Results:
Ang II caused expression of IFNg in C57BL/6 and STAT1-/- in vitro and in vivo. Blood pressure and cardiac hypertrophy did not differ between angiotensin treated C57BL/6 and STAT1-/- mice. Ang II increased in C57BL/6 expression of STAT1 dependent genes of chemokines and Nos2 (Cxcl10: 4.8-fold, Cxcl9: 3.4-fold, MCP-1: 6.6-fold, Nos2: 2.6-fold; all P<0.05) whereas this remained abolished in STAT1-/- mice. Ang II lead in STAT1-/- mice as compared to C57BL/6 to decreased cardiac CD45 number/view (C57BL/6-AngII: 27±4 STAT1-/--AngII: 11±5; P<0.05) and reduced cardiac fibrosis (hydroxyproline assay: C57BL/6-AngII: 80.6±11.8μmol/l versus STAT1-/- -AngII: 55.0±8.4μmol/l; P<0.05). Mesenteric arteries of STAT1-/- mice were fully protected against angiotensin induced endothelial dysfunction. Candesartan (AT1 receptor antagonist) inhibited action of Ang II in C57BL/6 and reduced but not reversed expression of chemokines in the heart.

Conclusions:
Lack of STAT1 revealed protection against Ang II-induced cardiac fibrosis and endothelial dysfunction. We suggest that STAT1-induced chemokine activation induces chemotaxis into hypertensive target organs and modifies cardiac fibrosis and vascular dysfunction in a blood pressure-independent manner.
Abstract 458: The Baroreflex Modulates The Anti-inflammatory Response Caused By Intravenous Lipopolysaccharide (lps)
Authors: Brognara Fernanda , Becari Christiane , Castania Jaci A , Martins Dias Daniel P , Tezini Geisa , Lopes Alexandre , Cunha Thiago M , Fazan Rubens , Salgado Helio C
Published: July 2021

Activation of the baroreflex elicits parasympathetic overactivity combined with sympathetic inhibition. The effect of electrical stimulation (ES) of the aortic depressor nerve (ADN) on the hemodynamic and inflammatory responses elicited by LPS (1.5 mg/kg iv) was examined in conscious rats. Rats were divided into 4 groups: 1) SALINE (n=6); 2) ES+SALINE (n=5); 3) LPS (n=7); 4) ES+LPS (n=5). Under ketamine/xylazine anesthesia the subjects were implanted with a pair of electrodes around the ADN and catheters into the femoral artery and jugular vein. On the next day, mean arterial pressure (MAP) and heart rate (HR) were recorded. ES (0.5mA; 0.25ms; 15Hz) was performed during 60 min after saline or LPS administration. Plasma was collected 1, 2 and 3h after saline or LPS administration, while the spleen and hypothalamus were collected after 3h to quantify interleukin (IL10) and tumor necrosis factor (TNFα). ES in the SALINE group promoted a prompt (5s) fall in MAP (-16± 5 mmHg) and HR (-39±12 bpm) followed by complete recovery of the MAP (7± 3 mmHg) and partial recovery of the HR (-8 ± 10 bpm) after 1h. In the LPS group ES promoted a prompt (5s) fall in MAP (-19± 6 mmHg) and HR (-40± 15 bpm) followed by complete recovery of the MAP (1 ± 3 mmHg) combined with significant tachycardia (30± 14bpm). Plasma TNFα increased 1h after LPS (400±78 vs 11±4 pg/μL from SALINE) followed by a decrease 2h (80±14 pg/μL) and 3h (34,4±5 pg/μL). After LPS the ES augmented IL-10 in the spleen (344±37 vs. 518± 79 pg/μL from ES+LPS) and decreased in the hypothalamus (73±8 vs 57±5 pg/μL from ES+LPS); while after saline ES increased IL-10 in the spleen (63±18 vs 314±99 pg/μL from ES+SALINE) but did not change in the hypothalamus (84±10 vs 97±17 pg/μL from ES+SALINE). After LPS administration ES did not change TNFα in the spleen (428±50 vs 450±98 pg/μL from ES+LPS) and hypothalamus (48±4 vs 38±5 pg/μL from ES+LPS); likewise in the spleen (285±38 vs 286±83 pg/μL from ES+SALINE) and hypothalamus (51,7±6 vs 54±8 pg/μL from ES+SALINE) after saline administration. These findings indicate that electrical activation of the baroreflex in conscious rats was capable to modulate the inflammatory response, promoting an increase of anti-inflammatory mediators in the LPS model of inflammation.
Abstract 476: Renal AT2 Receptor Activation Prevents Sodium Retention And Lowers Blood Pressure In Angiotensin II-Dependent Hypertension
Authors: Padia Shetal H , Howell Nancy L , Kemp Brandon A , Gildea John J , Keller Susanna R , Carey Robert M
Published: July 2021

A major proposed mechanism for the initiation of hypertension involves a primary increase in renal tubular sodium (Na+) reabsorption. Activation of intrarenal angiotensin type-2 receptors (AT2R) increases Na+ excretion; however, the role of intrarenal angiotensin type-2 receptors (AT2R) in the development of hypertension is unknown. Sprague-Dawley rats (N=36) underwent uninephrectomy and telemetric blood pressure probe implantation. Following a 72h recovery, two osmotic minipumps were inserted in each rat, one for chronic systemic delivery of 5% dextrose in water (D5W) or angiotensin II (Ang II, 200 ng/kg/min), and one for chronic intrarenal delivery of D5W (0.25 μL/h x 7d), highly selective AT2R agonist Compound 21 (C-21; 60 ng/kg/min x 7d), or specific AT2R antagonist PD-1223319 (PD; 10 ng/kg/min x 7d). Five groups of rats were studied: Group 1 (Control; N=10): systemic D5W + intrarenal D5W; Group 2 (Ang II-induced hypertension; N=8): systemic Ang II + intrarenal D5W; Group 3 (N=6): systemic Ang II + intrarenal C-21; Group 4 (N=6): systemic Ang II + 48h lead-in intrarenal C-21; Group 5 (N=6): systemic Ang II + intrarenal PD. Systemic Ang II infusion increased mean systolic blood pressure from 126±5 to 190±3 mm Hg over a 7d period in Group 2 (ANOVA F=73; P<1 X 10-6). Intrarenal administration of AT2R agonist C-21 (Groups 3 and 4) markedly inhibited the pressor effect of systemic Ang II (P<0.0001). Intrarenal AT2R antagonist PD (Group 5) augmented the pressor action of Ang II (P<0.0001). Consecutive 24h urinary Na+ excretion (UNaV) was reduced from 0.95±0.04 to 0.34±0.07 μmol/min (P<0.0001) on day 1 of Ang II infusion; Ang II-induced antinatriuresis was inhibited by intrarenal C-21 (P<0.0001) and augmented by intrarenal PD (P<0.0001) during the entire 7d infusion, demonstrating that one of the mechanisms to prevent Ang II-induced hypertension during intrarenal AT2R activation is the abolition of the initial increase in Na+ reabsorption that triggers the hypertensive cascade in this model. Thus, renal AT2Rs represent a novel therapeutic target for the prevention of hypertension.
Abstract 481: Deactivation of the renin-angiotensin-aldosterone system (RAAS) lowers glycated Hemoglobin (HbA1c) in hypertensive patients with Atherosclerotic Renal Artery Stenosis (ARAS)
Authors: Caielli Paola , Sanga Viola , Motta Raffaella , Battistel Michele , Calò Lorenzo , Maiolino Giuseppe , Miotto Diego , Rossi Gian Paolo
Published: July 2021

Background:
blockade of the RAAS lowered the incidence of Diabetes Mellitus in randomized clinical trials, but whether the de-activation of the RAAS can elicit the same beneficial effect in patients with renovascular hypertension (RVH) is unknown.

Aim:
to verify if endovascular treatment (EVT) could improve glycemic control in patients with atherosclerotic RVH.

Methods:
in the METRAS study (http://clinicaltrials.gov/show/NCT01208714, a randomized clinical trial comparing the effect of EVT on top of optimal medical therapy versus medical therapy alone (OMT) on GFR of the ischemic and contralateral kidney in patients with atherosclerotic RVH), glycemic control, as assessed by HbA1c at baseline and repeatedly during follow-up, was a pre-specified secondary endpoint of the study.

Results:
between June 2010 and March 2014, 16 patients were randomly assigned to EVT plus optimal medical therapy (n = 9) or OMT alone (n = 7). At baseline the 2 groups showed no significant difference of age, clinical and demographical features. All the patients, except two in the EVT group, were on a RAAS blocker. At 2 years follow-up HbA1c increased in patients on OMT (59±12 mmol/mol) from baseline values (45±16 mmol/mol, p<0.001). By contrast, in the patients assigned to EVT HbA1c remained stable (42±7 mmol/mol at baseline vs 41±5 mmol/mol at follow-up; p = NS; p<0.001 vs OMT at follow-up). When analyzed after multivariate adjustment for age, HbA1c at enrollment, and presence/absence of DM, the difference between the arms remained highly significant (p<0.001).

Conclusions:
in patients with atherosclerotic RVH and chronic activation of the RAAS, deactivation of the RAAS by means of EVT was associated with no increase in plasma levels of HbA1c over long term follow-up. At variance, similar patients assigned to optimized medical therapy alone showed worsened glycemic control over time. These findings support the contention of a causative role of RAAS activation in the incidence/progression of DM.
Abstract 484: The Vasorelaxing / No Release Effect Of Angiotensin-(1-9) Is Independent Of At2, Mas Or Mrgd Receptors
Authors: Resende Gabriel S , Silva Neyva , Villela Daniel C , Santos Robson A
Published: July 2021

Angiotensin-(1-9) is a nonapeptide formed by the hydrolysis of angiotensin I by ACE2 that seems to counter-regulate the classical RAS axis. Recent studies suggest that Ang-(1-9) acts via AT2 receptors (AT2R), however the pharmacological tool used to asses this ang-(1-9) /AT2R mediated interaction, is the AT2R antagonist, PD123319, that appears to have a great deal of inespecificity . Two other candidates for the ligation of Ang-(1-9), that have protective effects similar to the AT2R, are the MAS and the recently described MrgD receptor. Thus, in this study we addressed if Ang-(1-9) could be a ligand for AT2, MAS or MrgD receptors using aortic rings taken from AT2 and Mas knockout mice and AT2R or MrgD-transfected CHO cells.

Materials and methods:
The endothelium-dependent vasodilatory response to Ang-(1-9) was tested in aortic rings taken from Wild-Type, AT2KO and MASKO mice and Sprague-Dawley rats, pre-contracted with phenylephrine (0.1 umoles/L). NO release from AT2R or MrgD stable transfected CHO cells was evaluated using the NO indicator 4-amino-5 methylamino-2, 7 difluorofluorescein diacetate (DAF-FM) after Ang-(1-9) stimulation.

Results:
In aortic rings from SD rats Ang-(1-9) produced a dose-related relaxation (Emax= 15 ± 2.6) which was not modified by A-779 (Emax=12.9 ± 1.9) , the Mas/MrgD antagonist D-Pro7-Ang-(1-7) (Emax= 15.6 ± 1.7); or by PD123319 (Emax= 22.7 ± 5.4) . In aortic rings taken from MasKO there was a minor attenuation of the Ang-(1-9) vasorelaxant effect when compared to the WT, but the response was not abolished. No difference between AT2KO and WT mice was observed regarding the vasorelaxation produced by Ang-(1-9) (Emax= 20.7±2.4 and 20.9 ± 1.3 respectively). Moreover, the vasorelaxing effect of Ang-(1-9) was not affected by the association of PD123319 and D-Pro7-Ang-(1-7).

In addition, the nonapeptide did not stimulate NO production in AT2R-stably transfected CHO cells or MrgD stably transfected cells.

Taken together these results suggest that the vasorelaxation produced by Ang-(1-9) in rodent aortic rings is independent of Mas , AT2R or MrgD receptors.
Abstract 513: Impaired Hemodynamic Response to Head up Tilt in Adolescents Presenting with Chronic Nausea and Orthostatic Intolerance
Authors: Shaltout Hossam A , Wagoner Ashley L , Fortunato John E , Diz Debra I
Published: July 2021

We previously reported that ~70% of adolescents presenting to a Pediatric GI clinic for chronic nausea exhibit orthostatic intolerance (OI) in response to head upright tilt testing (HUT). The objective of this study was to determine whether supine mean arterial pressure or hemodynamic responses to HUT differ in these patients. Forty-eight patients (mean age of 15 [10-18] years, 36 females) completed a 45 minutes 0 to 70° HUT. Continuous blood pressure and heart rate recordings were acquired using non-invasive finger cuff. Thirteen subjects had normal tilt (Normal) while thirty five demonstrated OI. There were no differences between the two groups in supine blood pressures (BP), baroreflex sensitivity measured by frequency method in HF range (BRS), heart rate variability (HRV) measured as the root of mean square of successive differences (rMSSD), blood pressure variability (BPV) measured as standard deviation of mean arterial pressure (SDMAP) or the sympathovagal balance measure LF
RRI
/HF
RRI
. HUT caused a greater increase in heart rate in OI group (from 71 ± 6 beats/min to 104 ± 4 in OI vs from 75 ± 3 to 95 ±3 in normal, p=0.01) which was accompanied with lesser increase in BP (mainly due to lack of increase in diastolic) in the OI group. There was a trend for greater reduction in BRS in OI subjects (from 28.5 ± 13 ms/mm Hg to 6.3 ± 0.8 in OI vs from 21.1 ± 3.6 to 12.0 ± 2.9 in normal, p=0.09). HUT impaired HRV in both groups compared to supine values but the reduction was greater in OI group (-66.7 ± 4 % vs -52.0 ±5.6 in normal, p=<0.001). SDMAP increased by HUT compared to supine but to a greater extent in OI (40.6 ± 4 % vs 13.4 ± 8 in normal, p=0.02). LF
RRI
/HF
RRI
increased to a greater magnitude in OI group with HUT (from 1.8 ± 0.8 to 6.8 ± 0.8 in OI vs from 1.14 ± 0.18 to 4.1 ±0.7 in normal, p=0.02). These data reveal that the adolescents with orthostatic intolerance have attenuated parasympathetic responses and exaggerated activation of the sympathetic system to the heart and blood vessels. Despite these responses, subjects fail to maintain BP. Similar to previous studies in other subjects with OI, the excessive tachycardia often followed by syncope in most of these adolescents may reflect a loss of vascular responses to the activation of sympathetic and neurohumoral stimuli. Support: AHA12CRP9420029
Abstract 517: Ubiquitylation of G Protein Coupled Receptor Kinase 4 (GRK4) Regulates its Localization and Abundance
Authors: Gildea John J , Omer Walid , Bigler Wang Dora , Tran Hanh , Felder Robin A
Published: July 2021

The dopamine-1 receptor (D
1
R) is responsible for regulating up to 60% of natriuresis in the kidney under sodium loaded conditions. G protein coupled receptor kinase type 4 (GRK4) phosphorylates the D
1
R and reduces its membrane expression, but little is known about regulation of GRK4. We hypothesized that GRK4 is targeted for elimination through ubiquitylation. Using D
1
R and GRK4γ stably transfected human embryonic kidney cells (HEK293), we demonstrated GRK4 and ubiquitin coimmunoprecipitation. Addition of clasto-lactacystin beta-lactone (CLBL, an inhibitor of the 26S proteasome) increased GRK4 expression levels (western blot) using two different GRK4 fusion protein constructs. The addition of CLBL (10 μmol/L, 24 hrs) increased the expression of both a tandem affinity tagged (tap-tag) GRK4 fusion construct (2.65±0.19 fold over vehicle (VEH): VEH 4,010±404 RFU; CLBL 10,626±744 RFU, P<0.05, N=3) as well as a mCherry GRK4 fusion construct (6.98±0.50 fold increase over VEH: VEH 5,044±3229 RFU; CLBL 35,210±2547 RFU, P<0.05, N=3). The ubiquitin binding sites (AA 216,217; MYACK
Ub
K
Ub
LQKK) are located near the nuclear localization signal (AA 219-228; QKKRIKKRK) in GRK4. We therefore examined the subcellular localization of GRK4 after CLBL treatment. Nuclear accumulation of the GRK4-mCherry fusion protein fluorescence decreased markedly while cytoplasmic GRK4-mCherry fluorescence increased (nuclear to cytoplasmic GRK4 ratio: VEH 1.18±0.09 RFU; CLBL 0.78±0.03 RFU, P<0.01, N=12). In summary, GRK4 expression levels are negatively regulated by the ubiquitin proteasome system and ubiquitylation of GRK4 prevents nuclear import. We hypothesize that ubiquitylation is sterically hindering the nuclear localization sequence in GRK4 from interacting with the nuclear import machinery of the cell. Through manipulation of GRK4 ubiquitylation, one may be able to separate nuclear vs. cytoplasmic activity of GRK4 and reverse the negative impact overactive GRK4 has on dopaminergic signaling.
Abstract 541: Adenosine Inhibits C-Kit+ Progenitor Smooth Muscle Cell (P-SMC) Infiltration In Injury-Induced Neointima Formation And Human P-SMC Growth: Role of A2B Adenosine Receptor-Mediated Growth Regulation
Authors: Dubey Raghvendra K , Gillespie Delbert G , Fingerle Juergen , Imthurn Bruno , Jackson Edwin K , Baruscotti Isabella , Rosselli Marinella
Published: July 2021

Adenosine inhibits injury-induced neointimal formation; however, the mechanism is incompletely understood. Marrow-derived C-Kit+ progenitor cells (P-SMCs) express a smooth muscle phenotype and contribute to injury-induced neointimal thickening. The goal of our investigation was to determine whether adenosine attenuates neointimal hyperplasia in part by inhibiting the growth of P-SMCs and to clarify which adenosine receptor subtypes are involved. Digitized pixel analysis demonstrated that administration of adenosine (20 μM) peri-arterially for 7 days in a slow-release formulation inhibited injury-induced neointimal hyperplasia in the rat carotid artery by 71% (p<0.05: neointimal thickness: 37,424 ± 18,371 pixels in vehicle-treated [n=7] versus 10,352 ± 2,824 pixels in adenosine-treated [n=7]). Microscopic analysis following immunostaining for c-Kit+ cells showed a marked decrease (> 50%) in staining for c-Kit+ cells in the neointima of rats receiving adenosine. In cultured human P-SMCs, serum-induced growth (as determined by quantification of DNA synthesis and cell number), cell-migration, and collagen synthesis was inhibited by adenosine and 2-chloroadenosine, but not by N
6
-cyclopentyladenosine (selective A
1
-receptor agonist), CGS21680 (selective A
2A
-receptor agonist) or N
6
-(3-iodobenzyl)adenosine-5'-N-methyluronamide (selective A
3
-receptor agonist). This agonist profile was consistent with an A
2B
-receptor-mediated effect. Augmentation of endogenous adenosine levels with

erythro
-9-(2-hydroxy-3-nonyl)adenine (adenosine deaminase inhibitor)

and 5-iodotubericidin (adenosine kinase inhibitor) attenuated P-SMC growth. Moreover the antimitogenic effects of both exogenous and endogenous adenosine were abrogated by blockade of A
2B
receptors (MRS 1754), but not by antagonism of A
1
(8-cyclopentyl-1,3-dipropylxanthine), A
2A
(SCH 58261) or A
3
(VUF 5574) receptors. We conclude that adenosine, via activation of A
2B
receptors, inhibits neointimal formation in part by inhibiting infiltration and growth of C-Kit+ P-SMCs. These findings suggest that A
2B
receptor stimulation may prevent vascular remodeling associated with coronary artery disease, hypertension, atherosclerosis and restenosis.
Abstract 557: Blood-Pressure Lowering Effects of Canagliflozin Across a Range of Background Antihyperglycemic Treatment Regimens Used in Patients with Type 2 Diabetes Mellitus
Authors: Weir Matthew R , Yee Jacqueline , Vijapurkar Ujjwala , Meininger Gary
Published: July 2021

Hypothesis/Research overview:
Hypertension is a common comorbidity in patients with Type 2 Diabetes Mellitus (T2DM) and is an important risk factor for cardiovascular disease. Therefore, lowering blood pressure in patients with T2DM without meaningfully increasing heart rate is of benefit in reducing the risk of cardiovascular complications. In this analysis, the effect of the SGLT2 inhibitor canagliflozin (CANA) on blood pressure and heart rate was studied in patients with T2DM across a broad range of antihyperglycemic treatment regimens.

Methods:
In four Phase 3 clinical trials and two substudies, CANA 100 mg and 300 mg was compared to placebo (PBO) as monotherapy, dual therapy (add-on to metformin [MET] or sulfonylurea [SU]), triple therapy (add-on to MET + SU or MET + pioglitazone [PIO]), and add-on to insulin (± other antihyperglycemic agents). Efficacy and safety endpoints reported here include change from baseline in systolic blood pressure (SBP), diastolic blood pressure (DBP), and heart rate. Last observation carried forward imputation method was employed.

Results:
Treatment with CANA 100 mg and 300 mg was associated with reductions in SBP and DBP, with no change in mean heart rate, across the range of treatment regimens evaluated. PBO-subtracted changes (least square mean change from baseline and 95% confidence intervals) are reported for each endpoint (Table).

Conclusions:
Cana reduces blood pressure without changing heart rate whether used alone, in combination with oral antihyperglycemic agents, and with insulin in patients with T2DM.
Abstract 570: Characteristics of Long Non-coding RNAs in the Brown Norway Rat and Alterations in the Dahl Salt-Sensitive Rat
Authors: Wang Feng , Li Liping , Xu Haiming , Liu Yong , Yang Chun , Cowley Allen W , Wang Niansong , Liu Pengyuan , Liang Mingyu
Published: July 2021

Long non-coding RNAs (lncRNAs) are emerging as important mediators of genomic regulation. lncRNAs, however, remain poorly characterized in the rat model organism, which is widely used in hypertension research. Using poly(A)-independent and strand-specific RNA-seq, we identified approximately 1,500 to 1,800 lncRNAs expressed in each of the following tissues of Brown Norway rats: the renal cortex, renal outer medulla, liver, cardiac left ventricle, adrenal gland, and hypothalamus. Expression patterns and the binding of histone H3K4me3 to promoter regions were confirmed for several of the identified lncRNAs. Patterns of rat lncRNA expression appeared to be more tissue-specific than mRNA. Rat lncRNAs, on average, had similar lengths to mRNA but substantially fewer exons. The median abundance of rat lncRNAs was approximately 40% of that of mRNA. Approximately 28% of the lncRNAs identified in the renal outer medulla appeared to lack a poly(A) tail. Differential expression of 74 lncRNAs was detected in the renal outer medulla between Dahl salt-sensitive SS rats and salt-insensitive, congenic SS.13BN26 rats fed a high-salt diet. Two of the differentially expressed lncRNAs, which were confirmed, were located within the substituted congenic region. The study identified genome-wide characteristics of lncRNAs in the rat model and suggested a role of lncRNAs in hypertension.
Abstract 648: Nebivolol and Valsartan Increase Nitric Oxide Release from Human Endothelial Cells in a Synergistic Fashion
Authors: Mason R. Preston , Jacob Robert F , Malinski Tadeusz
Published: July 2021

Introduction:
Nebivolol is a β1-adrenergic receptor antagonist that stimulates endothelial nitric oxide (NO) release through β3-adrenergic receptor activation, ATP-mediated stimulation of purinergic P2Y receptors, and inhibition of membrane lipid oxidation. Valsartan is an angiotensin II receptor blocker (ARB) that selectively inhibits angiotensin II type 1 (AT1) receptors, thereby exerting no direct effect at type 2 (AT2) receptors, which have been shown to stimulate NO synthase activity through a bradykinin-mediated pathway. As NO is a key regulator of blood pressure and these two antihypertensive agents promote NO release through distinct mechanisms, we compared their individual and combined effects on NO release from human endothelial cells.

Methods:
Human umbilical vein endothelial cells (HUVECs) were incubated for 1 hr with vehicle, nebivolol or valsartan alone (each at 1.0 μM), or nebivolol (1.0 μM) and valsartan (0.5-5.0 μM) combined. The comparative effects of these agents on maximal NO release were measured in individual cells using porphyrinic nanosensors following stimulation with calcium ionophore (1.0 μM).

Results:
Nebivolol treatment increased HUVEC NO release by 49% (509 ± 18 nM, mean ± SD) as compared to vehicle treatment alone (342 ± 26 nM; p<0.001). Valsartan had a more modest effect, increasing NO release by 13% (385 ± 19 nM) as compared to vehicle-treated controls (p<0.01). Treatment with both agents at 1.0 μM increased HUVEC NO release by 91% (655 ± 19 nM) as compared to vehicle alone (p<0.001) and was 29% (p<0.001) and 70% (p<0.001) greater than the separate effects observed for nebivolol and valsartan, respectively. The additive effect of valsartan was dose-dependent and was also observed at 0.5 and 5.0 μM in combination with nebivolol.

Conclusions:
These data suggest that nebivolol and valsartan, when applied in combination, increase the ability of endothelial cells to release NO in a synergistic manner. The exact mechanism of this process remains unclear but, considering the importance of NO in regulating blood pressure, merits further study.
Abstract 660: Prevalence of Hypertension Among Children with Diabetes Mellitus.
Authors: Drutel Robert O , Paulo Remberto
Published: July 2021

This study aims to determine the prevalence of hypertension among children with diabetes mellitus, and describe adherence to the standard of practice regarding early diagnosis and treatment of hypertension in this population. Diabetes renders higher susceptibility to cardiovascular disease in affected patients and thus early detection of hypertension can be beneficial for future quality of life. We hypothesize that a number of diabetic children with hypertension are not promptly diagnosed and treated due to the difficult process preceding diagnosis. This cross-sectional study in an out-patient clinic of a university hospital was based on a group of 263 children (both females and males, age range between 3-18) diagnosed with type 2 diabetes. At the visit, the child's weight and height were recorded, BMI was calculated, systolic and diastolic blood pressure (BP) was taken, and blood sample drawn for hemoglobin A1C (HbA1C) readings. Within the study population we have identified a group of 73 children with hypertension. Comparing the two groups: the normotensive with the hypertensive, we have observed that the normotensive patients have significantly lower average BMI (26.7), p=0.000012 and slightly reduced HbA1C (8.9%), p=0.28 levels compared to the hypertensive group: BMI (32.5) and HbA1C (9.3%), respectively. We investigated the same parameters within every age group starting from age 10, and recorded that HbA1C was only significantly different for the group of 14 year-olds (8.7%; 11.7%, p=0.039). We also found that a significantly higher BMI is linked with hypertension for groups: age 13: BMI (29.2; 33.9, p=0.047), age 14: BMI (24.1; 35.6, p=0.00007) and age 18: BMI (31.8; 45.6, p=0.045). Within the different age groups there were differences between normotensive and hypertensive patients in BMI and HbA1C measurements, but they were not statistically significant and we assume that an increased sample size would be needed to confirm the data. We are currently working on identifying other risk factors including sex, race, height, urine creatinine, urine microalbumin, serum creatinine, lipid profile and thyroid function, that might be responsible for hypertension in pediatric patients with type 2 diabetes.
Abstract MP07: Nicotine Promotes Vascular Inflammation And Endothelial Dysfunction In Obese Rats
Authors: Zhou Ming-Sheng , Liu Chang , Chadipiralla Kiranmai , Tian Runxia , Raij Leopoldo
Published: July 2021

Epidemiological studies have shown that obesity and cigarette smoking (CS) are major cardiovascular (CV) risk factors and when coexisting in the same individuals have additive/synergistic effects upon CVD development. We have shown that stable compounds of CS (ATVB 2004) as well as nicotine, in concentrations found in smoker’s plasma, promote atherosclerotic CVD (AJP 2013). Here we studied in Sprague Dawley (SD) rats with diet -induced obesity the mechanisms involved in nicotine enhancement of CVD. SD rats (N=6-7 each group) were fed either a high fat (HF) or a standard chow (SCH) diet with or without nicotine (100 mg/kg/day in the drinking water) for 20 weeks. The HF rats developed central obesity, characterized by increased body weight gain (22%) and abdominal fat weight (53%), increased plasma levels of cholesterol (33%), non-esterified free fatty acids (68%), insulin (15%) glucose (12%)and systolic blood pressure (SBP: 146 ± 5 vs. 131 ± 5 mmHg SCH rats, p<0.05). Nicotine further increased SBP in obese HF rats (158 ± 4 mmHg, p<0.05) but not in lean SCH rats. Nicotine significantly increased O2- production in both obese (1689 ± 87 count/min/mg) and lean rats (1074 ± 105 count/min/mg) and further impaired EDR (Emax: 74 ± 5%) in obese HF rats. Nicotine also increased the expression of the macrophage marker ED1 in the aortas of obese HF rats. In peritoneal macrophages from obese HF rats TNFα, IL1β and CD36 were increased, and were further significantly increased in nicotine-treated obese HF rats. Using PCR array for inflammatory cytokines and receptor signaling pathway we found that the aortas from obese HF rats showed 2-4 fold increases in expression of several chemokines and interleukin genes expression; nicotine further increased the expression of 11 pro-inflammatory genes in the aortas from obese HF rats. Our results suggest that nicotine dramatically aggravates the CV effects of diet -induced obesity by increasing oxidative stress, vascular inflammation and endothelial dysfunction. Clinically chronic inhalation of nicotine, as that delivered by E-cigarettes, by individuals unable to quit the habit may have an important pro-atherogenic effect, particularly in obese subjects.
Abstract 004: Splenectomy Differentially Affects Angiotensin-2 and L-NAME Murine Models of Hypertension
Authors: Gigliotti Joseph , Cechova Sylvia , Le Thu
Published: July 2021

The immune system plays a major role in animal models of hypertension (HTN) and end-organ damage. However, few studies have assessed the role of lymphoid organs in the pathogenesis of HTN. We have shown previously that prior splenectomy (SPLX) significantly alters tissue inflammation; however the effect of SPLX on HTN remains unclear. Therefore, the objective of the current study is to determine whether prior SPLX influences the development of HTN in 2 different mouse models. Mice underwent SPLX or sham surgery 7 days prior to the induction of HTN using angiotensin-II (AngII, 400ng/kg*min s.c.) or nitric oxide synthase inhibition using L-NAME (30mg/kg*d in drinking water). Systolic blood pressure (SBP) was measured by tail-cuff manometer daily and mice were euthanized 14 days after induction of HTN. Heart weight/body weight (H/BW) ratios were calculated and kidney leukocyte infiltration was analyzed by flow cytometry. Mice with prior SPLX+AngII had significantly lower (P=0.03) SBP at both week 1 (148±7) and week 2 (135±7) as compared to Sham+AngII (174 and 173±7mmHg, n=5). Similarly, SPLX+AngII mice had significantly smaller (P=0.007) H/BW (4.3±0.3) as compared to Sham+AngII treated mice (5.2±0.4mg/g BW). Interestingly, no difference was observed in renal CD45+ (9.8±3 vs 10.6±3x105 cells/g, P=0.64) or CD3+ T-cell infiltration (8.8±0.2 vs 9.6±0.1x104 cells/g , P=0.64) between the Sham+AngII and SPLX+AngII treated mice, respectively (n=4/5). Furthermore, SPLX did not appear to influence the development of L-NAME HTN. SPLX+L-NAME mice had similar (P=0.84) SBP (145±4mmHg) as the Sham+L-NAME group (146±4mmHg, n=6) after 2 weeks. Relative heart weights were also similar (P=0.45) between SPLX+L-NAME (4.9±0.2) and Sham+L-NAME treated mice (4.8±0.3mg/g BW). Our data suggests that the full pressor response to AngII is dependent on the spleen. However, the effect of the spleen appears to be independent of renal inflammation. Moreover, the protective effect of the spleen is specific to AngII-dependent HTN and does not appear to be generalizable to all mouse models of hypertension. Further studies are needed to understand the physiological link between lymphoid organs (such as the spleen), renal inflammation, and the development of chronic HTN.