bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 184: Deterioration of Kidney Function by the (Pro)renin Receptor Blocker Handle Region Peptide in Aliskiren-treated Diabetic Transgenic (mRen2)27 Rats
Authors: Batenburg Wendy W , van den Heuvel Mieke , van Esch Joep H , van Veghel Richard , Garrelds Ingrid M , Bhaggoe Usha , Bouhuizen Angelique M , Leijten Frank P , Danser A H
Published: March 2022

Objective
Renal function in patients with diabetes mellitus (DM) might be influenced by the level of prorenin due to its binding to the (pro)renin receptor ((P)RR) and the subsequently occurring angiotensin production and/or the direct agonistic effects of prorenin mediated via this receptor. Indeed, in DM elevated prorenin levels are correlated with the development of microvascular complications such as nephropathy.

Design and Methods
In this study we evaluated renal function in diabetic TGR(mREN2)27 rats (a high prorenin hypertensive model), treated with vehicle, the renin inhibitor aliskiren, or aliskiren plus the putative (P)RR antagonist HRP for 3 weeks. 24-hour urine was collected, and blood and kidney were evaluated for renin-angiotensin system components and pathology.

Results
Increased diuresis and proteinuria due to DM were prevented by aliskiren, but not aliskiren+HRP. Aliskiren+HRP additionally decreased creatinine clearance, and increased the plasma levels of the profibrotic marker plasminogen-activator inhibitor-1. The increased natriuresis and renal collagen-1 expression in this model were unaffected by aliskiren±HRP. Aliskiren increased rat renin expression in the renal cortex. This was associated with a decline in (P)RR and AT
1
receptor mRNA expression, and these changes were unaffected by HRP. Glomerular volume and interlobar arterial lumen diameter modestly increased in the aliskiren+HRP group, and were unaffected by aliskiren alone.

Conclusions
HRP, when given on top of aliskiren in DM TGR(mREN2)27 rats worsens kidney damage and counteracts the beneficial effects of aliskiren. (P)RR blockade is therefore contraindicated in DM.
Abstract 186: ACE2 Activation Protects Corpus Cavernosum Impairment Induced by Hypercholesterolemia
Authors: Fraga-Silva Rodrigo A , Faye Younoss , Costa-Fraga Fabiana P , Shenoy Vinayak , Montecucco Fabrizio , Mach François , da Silva Rafaela F , Raizada Mohan K , Santos Robson A , Stergiopulos Nikolaos
Published: March 2022

Erectile dysfunction (ED) is multicausal disorder strictly associated with cardiovascular diseases. ACE2 is a key enzyme of the renin angiotensin system which plays an essential role in the cardiovascular system; however, the function of this enzyme in erectile tissues has never been investigated. Here we examined the expression of ACE2 in erectile tissues and its protective actions against hypercholesterolemic-induced corpus cavernosum injures. ACE2 expression was evaluated by immunostaining and western blot. Apolipoprotein E knockout (ApoE-/-) mice fed with Western-type diet for 11 weeks were treated with DIZE, an ACE2 activator compound, or vehicle during the last 3 weeks. Collagen and reactive oxygen species (ROS) content within the corpus cavernosum (CC) were evaluated by Sirius red and dihydroethidium staining, respectively. Protein expression of nNOS, eNOS, NADPH subunits (p67-phox and p22-phox) was assessed by western blot. Functional studies in CC strips were performed in an isometric organ bath. ACE2 was expressed in smooth muscle and endothelial cells of mouse CC. Interestingly, this enzyme was downregulated in penis from hypercholesterolemic mice. The treatment with DIZE reduced ROS production and NADPH oxidase expression in penis from ApoE-/- mice. Furthermore, DIZE treatment elevated vascular NO production (untreated-ApoE-/-: 11.3±1.1 vs. treated-ApoE-/-: 18.7±4.5, mol/L of serum NO3-) and augmented the expression of nNOS and eNOS in the penis. The NO-dependent relaxation induced by acetylcholine (ACh) was significantly increased in CC strip from DIZE-treated ApoE-/- mice (wild type 53.7%; untreated-ApoE-/- 39.3%; treated-ApoE-/- 52.1%, percentage of relaxation at 1μM of ACh). Additionally, DIZE treatment decreased collagen content within the CC. Of note, these beneficial actions of DIZE on the CC were not associated with changes in atherosclerotic plaque deposition or serum lipid profile. In summary, ACE2 activation attenuated oxidative stress, augmented NO bioavailability and decreased collagen deposition in CC of hypercholesterolemic mice. These results demonstrate that ACE2 is expressed and functional in erectile tissue, suggesting that this enzyme might have significant therapeutic benefits against ED.
Abstract 194: Low Levels of Angiotensin Converting Enzyme (ace) Do Not Prevent From Metabolic Alterations And Endothelial Dysfunction of Resistance Arteries Induced by High Fructose Intake
Authors: Moraes-Silva Ivana C , Irigoyen Maria-Claudia , Casarini Dulce E
Published: March 2022

Purpose:
ACE gene polymorphism may be associated with different responses to cardiovascular and metabolic disturbances. Therefore, we tested whether mice with low vs. normal levels of circulating and tissue ACE differently responds to chronic and excessive fructose (F) consumption.

Methods:
Male mice (22±1g) harboring 1 or 2 copies of the ACE gene received F (100mg/l) or tap water for 8 weeks. Blood pressure (BP) was measured by plethysmography. Glycemia, lipids and glucose tolerance (0 to 120 min after glucose load=1.5 g/Kg ip) were also evaluated. After euthanasia, the white adipose tissue was weighed and the mesenteric resistance artery (MRA) was dissected out. Relaxation of MRA rings was evaluated in an isometric myograph in response to acetylcholine (Ach;0.01nM-30μM) or sodium nitroprusside (SNP; 0.01nM-300μM) after pre-constriction with noradrenaline.

Results:
Caloric intake (~ 12.7 kcal/day) and body weight gain (1 copy: ~40%; 2 copies: ~30%) were similar among all groups. However, adipose tissue deposition was 43% higher in 1-copy F-fed mice and 36% higher in 2-copy F-fed mice in relation to the controls (p<0.05), with no influence of ACE gene copy number. F consumption increased systolic BP in 2-copy mice (1 copy: 114±2 vs. F=116±2; 2 copies: 113±2 vs. F=122±1 mmHg) while diastolic BP was increased in both F-fed groups (1 copy: 79±1 vs. F= 88±2; 2 copies: 81±2 vs. F=93±2 mmHg). This augment in BP was significantly higher in 2-copy than in 1-copy mice (p<0.05). F-fed mice also presented hyperglycemia (1 copy: 84±3 vs. F=128±5; 2 copies: 85±4 vs. F=133±8 mg/dl) as well as glucose intolerance (increase of ~67% in the area under time course curve); with no alterations in total cholesterol nor triglycerides. MRA % of relaxation was unaltered in response to SNP, while F-fed groups showed reduced relaxation response to Ach (1 copy: 87% vs. F=51%; 2 copies: 92% vs. F=57%).

Conclusions:
F intake provokes glucose intolerance, increases visceral adipose tissue deposition and decreases endothelium-dependent relaxation of resistance arteries at a similar magnitude for both, 1- and 2-copy ACE gene mice, while F-induced BP increase seems to be more dependent on ACE gene dosage.
Abstract 195: Circadian Rhythm of Plasma and Urinary Angiotensinogen in Patients with Chronic Kidney Disease
Authors: Nishijima Yoko , Kobori Hiroyuki , Mizushige Tomoko , Hara Taiga , Nishiyama Akira , Kohno Masakazu
Published: March 2022

Recent basic and clinical data demonstrated that the intrarenal renin-angiotensin system (RAS) plays an important role in the progression of chronic kidney disease (CKD). The urinary angiotensinogen (AGT) excretion rate could be a novel biomarker for the activity of the RAS in the kidney. We previously reported that the healthy volunteers do not have a circadian rhythm of AGT level in urine (Hypertension. 2011;57:e78) or in plasma (Hypertension. 2012;60:A399). However, the circadian rhythm of AGT level in urine and in plasma in patients with CKD has not been reported yet. Therefore, this study was performed to investigate the circadian rhythm of AGT level in urine and in plasma in patients with CKD.

We recruited 6 CKD patients with continuous proteinuria admitted to the Kagawa University Hospital from 06/2011 to 10/2011 for the purpose of diagnostic renal biopsy. Plasma samples were collected at 06:00, 12:00, and 18:00. Urine samples were collected at 06:00, 09:00, 12:00, and 18:00. Plasma renin activities (PRAs), plasma and urinary AGT concentrations, and urinary albumin (Alb) concentration were measured using commercially available kits. The urinary concentrations of AGT and Alb were normalized by the urinary concentration of creatinine (Cr) (UAGT/Cr and UAlb/Cr, respectively).

PRA (2.07 +/- 0.77 ng of angiotensin I/mL/hr at 06:00, 2.45 +/- 0.77 at 12:00, and 2.58 +/- 0.76 at 18:00, P = 0.8853) or plasma AGT (19.9 +/- 2.4 μg/mL at 06:00, 24.1 +/- 3.1 at 12:00, and 23.0 +/- 3.9 at 18:00, P = 0.6300) did not show a circadian rhythm. Moreover, UAlb/Cr (511 +/- 319 mg/g Cr at 06:00, 827 +/- 459 at 09:00, 1479 +/- 862 at 12:00, and 1370 +/- 818 at 18:00, P = 0.6964) or UAGT/Cr (54 +/- 22 μg/g Cr at 06:00, 119 +/- 41 at 09:00, 350 +/- 194 at 12:00, and 198 +/- 104 at 18:00, P = 0.3035) did not show a circadian rhythm.

In conclusion, in addition to healthy volunteers, patients with CKD do not have a circadian rhythm of AGT level in urine or in plasma.
Abstract 197: Effects of a Stabile Angiotensin-(1-7) Analogue, Cyclic Angiotensin-(1-7), on Progenitor Cell Recruitment and Cardiac Remodeling
Authors: Sevá Pessôa Bruno , van Veghel Richard , Danser A.H. Jan , Roks Anton J.M.
Published: March 2022

Objectives:
In animal models, angiotensin-(1-7) treatment has beneficial effects improving cardiac function and remodeling after myocardial infarction (MI). Angiotensin-(1-7) might have a beneficial angiogenic effect after MI, since infusion increases the number of progenitor and VEGF+ cells in the heart. Angiotensin-(1-7) also promotes the
in vitro
differentiation of endothelial progenitor cells (EPC), and tube formation. For optimal clinical use, metabolically stable 4-Ser,7-Cys-thioether-bridged angiotensin-(1-7), called cyclic angiotensin-(1-7) (cAng-(1-7)), has been developed. We studied the effect of cAng-(1-7) treatment on progenitor cells recruitment and early cardiac remodeling after MI, with an emphasis on angiogenic properties.

Design and Methods:
Angiogenic progenitor cell recruitment was measured 24 h after a bolus injection of 50 ug/kg cAng-(1-7) in healthy mice by flow cytometric counting of ckit+, sca-1+ and flk1+ cells in blood and bone marrow. In addition, mice underwent coronary ligation or sham surgery and either saline or 2.4 ug/kg/h cAng-(1-7). After 3 weeks of treatment, cardiac function was measured by intraluminal pressure catheter. Cardiac vascular density, fibrosis and myocyte dimensions were assessed. Furthermore, we studied the dose-related effects of normal and cAng-(1-7) on tube formation by HUVEC.

Results:
cAng-(1-7) increased blood ckit+, sca-1+ and flk1+ cells, which tended to decrease in bone marrow. cAng-(1-7) reduced cardiomyocyte hypertrophy (476±99μm
2
vs
292±78μm
2
/cell) compared to saline, but showed no effect on heart weight, infarct size, fibrosis and vascular density. Cardiac contractile function was not improved by cAng-(1-7). Tube formation by HUVEC was reduced by cAng-(1-7) as well as by native Ang-(1-7).

Conclusions:
Our results suggest that cAng-(1-7) given early after MI might not lead to improved angiogenesis, despite an increased recruitment of hematopoietic and endothelial-like cells from the bone marrow. This might explain why there is no swift beneficial effect of cAng-(1-7) on cardiac performance despite a reduction of myocyte hypertrophy. Further optimization of cAng-(1-7) treatment and evaluation of treatment lasting longer than 3 weeks will be necessary.
Abstract 2: Compensatory Activation of the Sympathetic Nervous System after Deletion of Angiotensin Type 1 Receptors from Vascular Smooth Muscle Cells
Authors: Sparks Matthew A , Stegbauer Johannes , Chen Daian , Gomez Jose A , Herrera Marcela , Gurley Susan B , Coffman Thomas M
Published: March 2022

Vasoconstriction is a signature physiological action of angiotensin II (Ang II). In order to define the physiological role of direct actions of AT1 receptors (AT1 R) in VSMCs, we generated mice with cell-specific deletion of AT1A R from VSMCs using a floxed Agtr1a allele and transgenic mouse lines expressing Cre only in VSMC lineages. We first compared systemic vascular responses to acute infusions of Ang II in mice lacking AT1A R in VSMCs (SMKOs) and controls. Surprisingly, brisk vasoconstrictor responses to 1μg/kg Ang II were preserved in SMKOs with peak increases in MAP only modestly reduced to ≈75% of control levels (15±2 vs. 23±1 mmHg P<0.05). Robust systemic vasoconstriction to Ang II was observed whether elimination of AT1A Rs was carried out in utero with a constitutively expressed SM-Cre or in adult animals with a tamoxifen-inducible SM-Cre. By contrast, Ang II-dependent vasoconstriction in the renal circulation was largely abrogated in SMKOs. Moreover, despite preserved systemic vascular responses to acute Ang II in SMKOs, their baseline BP was reduced and susceptibility to Ang II-dependent hypertension was significantly attenuated. To determine whether the minor AT1BR isoform might be responsible for their residual acute vascular responses, we generated SMKOs on an AT1B-null background (SMKO 1B-/-); the absence of AT1B Rs did not affect this response (SMKO 1B-/- 16±2 vs. SMKO-1B+/+ 15±2 mmHg; ns). As an alternative explanation, we tested for a possible involvement of the sympathetic nervous system (SNS) by infusing 400μg/kg of the α-adrenergic blocker phentolamine (PT) 5 min before giving Ang II. While PT had no effect on the intensity of Ang II-dependent vasoconstriction in controls (pre-PT: 23±1 vs. post-PT: 23±3 mm Hg), it almost completely abrogated vasoconstriction in SMKOs (pre-PT: 15±2 vs. post-PT: 3±1 mm Hg; P<0.005). Thus, we find a significant capacity for activation of the SNS to maintain acute Ang II-dependent vasoconstriction when AT1R signaling in VSMCs is absent. This pathway can induce brisk vasoconstriction in the systemic but not the renal circulation. Furthermore, despite compensatory activation of the SNS, deletion of AT1Rs from VSMCs reduces baseline BP and attenuates the severity of Ang II-dependent hypertension.
Abstract 200: Inhibition of TNFa-Induced ICAM-1 Expression by AT2 Activation Involves ACE2
Authors: Zhu Liping , Carretero Oscar A , Xu Jiang , Yang Xiao-Ping
Published: March 2022

The renin-angiotensin system (RAS) is a coordinated hormonal cascade involved in cardiovascular control, with angiotensin II (Ang II) as the main effector peptide. Both Ang II type 2 receptor (AT2) and angiotensin I-converting enzyme 2 (ACE2) are important in RAS system, and counteract the vasoconstrictor and pro-inflammatory effects of Ang II. Recent studies have suggested a link between AT2 and ACE2. However, the role of ACE2 in mediating the anti-inflammatory effects of AT2 has not been established. Tumor necrosis factor (TNFα) is a cytokine involved in Ang II signaling and promotes the inflammatory response
via
NF-kB. We hypothesized that activation of AT2 prevents TNFα stimulation of intercellular adhesion molecule-1 (ICAM-1) expression in human coronary artery endothelial cells (HCREC) through ACE2. Cultured HCREC were pretreated with AT2 antagonists PD123319 (10 μM) or ACE2 antagonist DX 600 (1 μM) for 30 min, then AT2 agonist CGP42112A (0.1 μM) for 30 min and then stimulated with TNFα (0.5 ng/ml). ICAM-1 and
p
-IkB expression were measured by Western Blot. ACE2 mRNA was measured by real-time RT-PCR. ICAM-1 expression was very low in untreated cells and greatly increased by TNFα treatment. Pretreatment with AT2 agonist CGP42112A inhibited TNFα-induced ICAM-1 expression by 47% ± 5% (
p
<0.01). This effect was diminished by AT2 antagonist PD123319. Interestingly, activation of AT2 with agonist CGP42112A increased ACE2 mRNA expression by 1.82 ± 0.09 fold (p<0.01). This increase was further enhanced by overexpression of AT2 in the presence of CGP42112A. Pretreatment with ACE2 antagonist DX 600 prevented the inhibitory effects of AT2 stimulation on TNFα-induced ICAM-1 expression (
p
<0.01), whereas DX 600 itself had no effect. Furthermore, we found that TNFα increased
p
-IKB expression by 7.46 ± 0.51 fold (
p
<0.01) compared to untreated cells and this effect was diminished by activation of AT2 with CGP42112A. Our findings suggest that stimulation of AT2 reduces TNFα-induced ICAM-1 expression, which is partly through ACE2. NF-kB may be involved in this signaling cascade.
Abstract 204: Sodium Sensing Network in Hypertension-Induced Cardiac Hypertrophy
Authors: Pinho Maria J , Ciscotto Paula , Bertorello Alejandro M , Soares-da-Silva Patricio
Published: March 2022

Hypertension is related to alterations of sodium homeostasis that promote abnormal accumulation of water in the intravascular compartment leading to blood pressure elevation. This process can been associated with increased heart mass resulting in cardiac hypertrophy. At cellular level, the concentration and active transport of Na+ is regulated by Na+,K+-ATPase enzyme (NK). In vitro studies have shown a correlation between high blood pressure, NK and SIK activities. Increases in intracellular Na+ are paralleled by elevations in intracellular Ca2+ via NCX1, leading to the activation of SIK1 by a Ca2+/calmodulin-dependent kinase. Activation of SIK1 results in the dephosphorylation of the NK α-subunit and an increase in its catalytic activity. Accordingly, the aim of the present study was to evaluate the cardiac Na+ sensing network changes associated with hypertension in aging hypertensive rats. Systolic and diastolic blood pressures, determined by the tail-cuff method, were significantly higher in 3, 13 and 21-month old SHR than in age-matched WKY. In the SHR, ageing was associated with increases in cardiac mass, gene expression of hypertrophic (α-skeletal muscle actin and β-myosin heavy chain) markers as well as inflammatory cytokine (IL-6). The decreased expression of heart SIK isoforms, SIK1 and SIK3, was associated with downregulation of transcription factors Snail2, Zeb1, MFAT5c and KLF4, in 13 and 21-month old SHR. Cardiac α2-isoform of NK was found to be decreased in 21 month old SHR. Whereas, NKα1, NHE3 and NCX1 showed no differences between age-matched SHR and WKY. Na+-dependent ASCT2 and Na+-independent LAT1 amino acid transporter transcript levels of were significantly higher in 21 month old SHR than in WKY. In the aged SHR, cardiac hypertrophy was associated with dysregulation of several elements of the intracellular Na+ sensing network. Downregulation of SIK1 and transcription factors is correlated with expression of NKα2, suggesting impaired sodium handling. It is proposed that, cardiac hypertrophy is not exclusively the consequence of mechanical stress but also of other factors associated with elevated blood pressure such as abnormal cell sodium homeostasis.Supported by grant PIC/IC/83204/2007.
Abstract 215: The Role of ER Stress in Renal Diseases in Metabolic Syndrome Animals : Blockade by Natural Antioxidants
Authors: R. Nair Anand , Francis Joseph
Published: March 2022

ER stress has been implicated in hypertension, obesity, insulin resistance and diabetes, all of which are hallmarks of metabolic syndrome (MetS). MetS is also characterized by an increased risk for cardio-renal diseases. Recent evidences demonstrate a role for ER stress in kidney injury. Recently we demonstrated that blueberry (BB) treatment attenuates renal injury in spontaneously hypertensive rats. However, the mechanism for this protective effect is not known. Since blueberries are known to have antioxidant and anti-inflammatory properties, we hypothesized that BB protect against renal dysfunction in MetS by attenuating ER stress.

Methods:
Male zucker rats - Lean controls (LZR) and Obese (OZR) were both fed with a BB-enriched diet or a control diet for 15 weeks. At the end of the feeding period, the rats were subjected to acute renal clearance experiments to determine the blood pressure and kidney function parameters. Kidney cortex tissues were examined for gene and protein expression of ER stress and kidney injury markers, by RT-PCR and western blotting respectively.

Results:
are tabulated. Further studies are being carried out to investigate whether blocking ER stress would attenuate the structural and functional kidney damage in MetS animal models.

Conclusions:
1) ER stress increased in the kidneys of MetS animals and contribute to renal dysfunction. 2) BB ameliorates the progression of renal diseases in MetS by attenuating ER stress, a novel therapeutic approach.
Abstract 216: Dietary Salt Restriction Improves Cardiac and Adipose Tissue Pathology Independently of Obesity in a Rat Model of Metabolic Syndrome
Authors: Hattori Takuya , Murase Tamayo , Takatsu Miwa , Ohura Sae , Takeshita Yuuri , Takahashi Keiji , Murohara Toyoaki , Nagata Kohzo
Published: March 2022

Introduction:
Metabolic syndrome (MetS) enhances salt sensitivity of blood pressure and is responsible for cardiovascular disease. However, few studies have reported the effects of dietary salt restriction on the link between cardiovascular and metabolic disorders in MetS. We investigated whether dietary salt restriction might ameliorate cardiac and adipose tissue injury in a rat model of MetS.

Methods and Results:
We used DahlS.Z-

Lepr
fa
/Lepr
fa

(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 fed a normal-salt (0.36% NaCl in chow) or low-salt (0.0466% NaCl in chow) diet from 9 weeks of age and were compared with homozygous lean littermates (DahlS.Z-

Lepr
+
/Lepr
+

, or DS/lean rats). DS/obese rats fed the normal-salt diet progressively developed severe hypertension and showed left ventricular (LV) hypertrophy, fibrosis, and LV diastolic dysfunction at 15 weeks of age. Dietary salt restriction markedly attenuated all of these changes in DS/obese rats. The levels of cardiac oxidative stress and inflammation and the expression of cardiac renin-angiotensin-aldosterone system genes were increased in DS/obese rats fed the normal-salt diet, and these parameters were down-regulated by dietary salt restriction in both DS/obese and DS/lean rats. In addition, dietary salt restriction suppressed the increases in visceral adipose tissue inflammation and circulating proinflammatory cytokines apparent in DS/obese rats. Body weight and visceral adipocyte size were not affected by dietary salt restriction. Dietary salt restriction did not alter fasting serum glucose levels, but it markedly reduced the fasting serum insulin concentration and the HOMA-IR in DS/obese rats.

Conclusions:
Dietary salt restriction not only prevents hypertension and cardiac injury but also ameliorates systemic and adipose tissue inflammation and insulin resistance, without affecting obesity, in this model of MetS.
Abstract 217: Chronic Treatment with Carbon Monoxide Releasing Molecule Reverses Dietary Induced Obesity in Mice
Authors: Hosick Peter A , Stec David E
Published: March 2022

We have recently demonstrated that chronic, low level treatment with carbon monoxide releasing molecules (CORMs) prevents the development of obesity in response to a high fat diet. The objective of this study was to test the hypothesis that chronic, low level treatment with CORM can reverse established obesity via a mechanism independent of food intake. This hypothesis was tested by treating 24 week old DIO C57BL6/J mice with a low level of the CO releasing compound, CORM-A1 (5 mg/kg body weight, i.p.; n=4) or the inactive compound iCORM-A1 (n=6) every 48-hours for 30 weeks while maintaining a high fat (60%) diet. Initial body weights (BW) were not different between the groups; however, at 30 weeks of treatment, CORM-A1 significantly reduced body weight as compared to iCORM-A1 (32.7±2.9 g (62% of initial BW) vs. 52.1±13.2 g; p<0.05 (98% of initial BW)). CORM-A1 treatment also reduced fasting glucose by 20-23% at 12, 18, 24, and 30 weeks compared to iCORM-A1 (p<0.05). Further, an intraperitoneal glucose tolerance test conducted at 25 weeks into treatment demonstrated that glucose utilization was improved by CORM-A1 treatment as area under the curve (AUC) was significantly reduced 15062±2568 AUC vs. 20795±1809 AUC; p<0.05). Oxygen consumption was increased in CORM-A1 versus iCORM-A1 treated mice (90.2±20.3 mL/kg
LBM
/min vs.70.4±16.6 mL/kg
LBM
/min; p=0.07). However, weight loss in CORM-A1 treated mice was independent of differences in motor activity (4665±2396 cm vs. 5339±3325 cm, p=0.81) and food intake (3.1±0.2 g/day vs. 3.5±0.1 g/day; p=0.1). These results demonstrate that chronic, low level treatment with a CO donor at levels which do not raise blood carboxyhemoglobin levels reverses established obesity in mice fed a high fat diet through a mechanism independent of food intake. Our results suggest that chronic, low level treatment with CO releasing compounds may be a novel therapy for the reversal of obesity. This work was supported by grants from the NHLBI, PO1HL-51971, HL088421 and 1T32HL105324.
Abstract 218: Age Impacts Circulating Testosterone Levels in Male Intrauterine Growth Restricted Rats Leading to Glucose Intolerance and Alterations in Programmed Hypertension
Authors: Intapad Suttira , Fahling Joel , Alexander Barbara
Published: March 2022

Low birth weight increases the risk for hypertension, obesity, and impaired insulin and glucose metabolism in later life. We previously reported that reduced uterine perfusion induced at day 14 of gestation in the pregnant rat leads to intrauterine growth restriction (IUGR) and programs testosterone dependent hypertension at 4 months of age in male IUGR offspring relative to male control. Whether age impacts chronic health in male IUGR is unknown. Thus, the aim of this study was to test the hypothesis that age impacts cardiovascular and metabolic health in male IUGR offspring. Male control and male IUGR offspring were studied at 6 and 12 months of age. Mean arterial pressure (MAP) measured in conscious, chronically instrumented animals via arterial catheter was significantly increased in IUGR at 6 months of age (138.9±2 mmHg, n=9;
P
<0.05) relative to age-matched control (127.4±4 mmHg, n=9). However, blood pressure was not significantly elevated at 12 months of age (IUGR: 142.9±4 mmHg, n=10 versus control: 136.7±2 mmHg, n=6). Glucose tolerance was maintained in IUGR at 6 months of age versus age-matched control as demonstrated by area under the curve (AUC) (456.2±28, n=7 versus 436.6±14, n=11; respectively). However, glucose tolerance was impaired in IUGR at 12 months of age (509.1±18, n=10;
P
<0.05) versus age-matched control (437.2±13 AUC, n=9). Insulin tolerance and insulin release in response to an oral glucose challenge were similar in IUGR and control at 12 months of age. Age-dependent increases in body weight, visceral fat, and total fat mass were also similar and 24 hour food intake and water intake did not differ at 12 months of age. Testosterone deficiency is reported to be associated with impaired metabolic health. In this study, circulating testosterone remained markedly higher in IUGR (448.9±156 ng/dl;
P
<0.05) at 6 months of age relative to male control (152.2±17 ng/dl); yet, was no longer significantly elevated at 12 months of age (IUGR: 256.6±40 versus control: 177.4±18 ng/dl). Thus, these findings indicate that male IUGR offspring exhibit glucose intolerance and a loss of programmed hypertension at 12 months of age that may be due to an age-dependent decrease in testosterone. Future studies are needed to investigate the exact underlying mechanisms.
Abstract 223: Relationship Between Leukocyte Count and Platelet Count as Markers of Inflammation and Obesity in Patients With Acute Chest Pain
Authors: Chaikriangkrai Kongkiat , Kassi Mahwash , Khaleel bala Sayf , Chang Su Min
Published: March 2022

Introduction
Obesity is associated with low-grade chronic inflammation which has been linked to complications of obesity including coronary artery disease. Epicardial adipose tissue volume (EATv), a surrogate of visceral adiposity burden, has been reported to be associated with adverse cardiovascular risk factor profiles. The aim of this study was to examine the relationship between leukocytes count and platelet count as markers of inflammation and obesity defined by body mass index (BMI) and EATv in patients with acute chest pain.

Methods
In this cross-sectional study, 1030 consecutive patients without reported history of coronary artery disease who presented with acute chest pain to our hospital were included. Patients with any signs of infection or inflammation were excluded. Cardiac risk, complete blood counts and EATv measured by non-contrast multi-detector computed tomography scan were collected. Patients were categorized by BMI classification and EATv quartiles. Data were described in mean and standard deviation. Students’t-test, ANOVA and linear regression analysis were used when appropriate.

Results
The population consisted of 21.2% (206 of 972) patients without overweight or obesity, 33.2% (323 of 972) with overweight, 45.6% (443 of 972). A subset of 72% (704 of 972) has EATv measured. Compared to non-overweight/non-obese group, obese group have similar leukocyte count (mean 7800±3330 vs 8200±7600; p-value=0.349) and platelet count (mean 253.07±71.51vs 240.96±79.25; p-value=0.056) but higher absolute lymphocyte count (mean 2180±1620 vs 1940±1840; p=0.04). Obese group also had higher absolute lymphocyte count compared to overweight group (2180±1620 vs 1940±1030; p=0.003). BMI is independently associated with absolute lymphocyte count in linear regression analysis (beta = 17). Among EATv quartiles, there was no significant difference in leukocytes count, absolute neutrophil count, absolute lymphocyte count and platelet count.

Conclusion
Neither leukocyte count nor platelet count is correlated with obesity defined by either BMI or epicardial adipose tissue volume. However, higher BMI is independently associated with higher absolute lymphocyte count in patients with acute chest pain.
Abstract 225: Relationship Between Blood Pressure Lowering Medication Profiles and Coronary Artery Disease in Patients With Acute Chest Pain
Authors: Chaikriangkrai Kongkiat , Kassi Mahwash , Khaleel Bala Sayf , Chang Su Min
Published: March 2022

Introduction
Whether use of blood pressure lowering medications (BPLMs) is associated with presence and severity of coronary artery disease (CAD) is unknown. The aim of this study was to examine the relationship between BPLMs and coronary artery calcium score (CACS) as a marker of CAD in patients with acute chest pain.

Methods
In this cross-sectional study, 1030 consecutive patients without reported history of CAD who presented with acute chest pain were included. CACS by non-contrast multi-detector computed tomography scan, BPLMs profiles prior to the visit were collected. Patients were categorized by presence and severity of CACS, number and classes of BPLMs used.

Results
The population with mean age of 54±13 years consisted of 60.6% (624 of 1030) patients with zero CACS, 21.7%(223 of 1030) with mild calcification(0<CACS<100) and 17.8%(183 of 1030) with moderate-to-severe calcification(CACS≥100). Patients with single BPLM and multiple BPLMs are accounted for 24% and 24% of all the patients respectively. Compared to non-BPLM group, BPLMs group had more patients with presence of CACS and moderate-to-severe calcification (238 of 494; 48.2% vs 168 of 536; 31.3% p<0.001; for CACS>0 and 118 of 494; 23.9% vs 65 of 536;12.1% p<0.001;for CACS≥100). Compared to single BPLM group, multiple BPLMs group had no significant difference in numbers of patients with presence of CACS and moderate-to-severe calcification (p=0.106 and p=0.345 respectively). Most BPLMs classes are associated with CACS as shown in table1.

Conclusion
Use of blood pressure lowering medications is associated with presence and severity of coronary artery disease in patients with acute chest pain.
Abstract 228: Circadian Patterns of Heart Rate Turbulence, Heart Rate Variability in Essential Hypertension
Authors: Tuyzarova Irina , Dimitriev Dimitri
Published: March 2022

Purpose.
The aim of this study was to investigate the effect of circadian change of autonomic tone on heart rate turbulence (HRT) and variability (HRV) in patients with essential hypertension and matched controls.

Methods.
In 25 essential hypertension patients (dippers) and matched 25 controls (mean age 54,83 +/- 1 years; 47,9% female) with at least 100 ventricular premature beats in 24-hour Holter monitoring, turbulence onset, turbulence slope, and heart rate variability parameters were calculated in 2 equal 12-hour periods during day and night.

Results.
There were 15% and 13% with abnormal turbulence onset (TO) and turbulence slope (TS) values in patients. No circadian changes in TO values were found, while TS displayed significantly lower values during day than during night hours ((mean±std. err.) controls: 8.07±0.68, vs. 12.29±0.9 ms/RR, respectively; patients: 5.11±0.89, vs. 9.04±0.97 ms/beat, respectively; p <0.05). Absolute value of daytime TO was significantly lower in the patients with hypertension compared to controls. (-0.95±0.2l% vs. -1.92±0.37% p <0.05) and daytime levels of TS were higher in controls (p<0.01). Nighttime TO and TS values were not significantly different between the patients and controls. Spectral analysis showed that power in the high-frequency range (0.15 to 0.40 Hz) was lower among the hypertensive patients during night (269.1±50.5 ms2 vs. 689.9±33.04 ms2, p<0.01). Power in the low-frequency range (0.04 to 015 Hz) was higher among the hypertensive patients at day (1697.5±89.2 vs. 769.9±57.9, p<0.01) and at night (2499.3±94.4 vs. 1703.4±60.5, p<0.01). The group mean cosinor analysis and the acrophase-amplitude analysis demonstrated significant differences in circadian rhythm in pLF, pHF and LF between the group with essential hypertension and controls. In analysis the relationship between HRT and HRV parameters, TS has higher correlation with HRV than TO values.

Conclusions.
Circadian change of TS values is more prominent than TO. Our findings indicate that hypertensive subjects were characterized with a decreased physiological circadian fluctuation on autonomic functions compared with controls.
Abstract 233: Resistance Training Attenuates Renal Dysfunction in Animal Diabetic Nephropathy but Does Not Restore Kidney ACE/ACE2 Balance
Authors: Santos Ralmony A , Silva Kleiton A , Aragao Danielle S , Arita Danielle Y , Rosa Rodolfo M , Arlotti Marcelo R , Firoozmand Lilia T , Peres Giovani B , Moraes Milton R , Schor Nestor , Casarini Dulce E , Cunha Tatiana S
Published: March 2022

Overactivity of the intrarenal renin-angiotensin system (RAS) has been implicated in the pathogenesis of diabetic nephropathy. Our group has already demonstrated that aerobic exercise reduces kidney angiotensin II levels and attenuates renal dysfunction under concurrence of diabetes and hypertension. Resistance training (RT) has recently been recognized as a useful therapeutic tool for the treatment chronic diseases and similar to aerobic exercise, has been reported to improve glycemic control. Therefore, the aim of this study was to evaluate the effect of RT on renal function and RAS in diabetic animals, to understand whether this type of exercise is also associated with renoprotection. Wistar rats (3 months old) were randomized into: sedentary control (SC); trained control (TC); sedentary diabetic (SD) and trained diabetic (TD). Animals were made diabetic with a single tail injection of streptozotocin (STZ, 50 mg/Kg). RT was performed on an 110-cm ladder (8 ladder climbs, once/day, 5 days/week, 8 weeks), carrying a load of 50-80% body weight (BW) appended to the tail. At week 8, 24 hr urine volume and albuminuria were evaluated. Kidney was excised and ACE and ACE2 activities were determined (ZPhe-HL and 7-Mca-APK(Dnp), respectively) (Two way ANOVA + Tukey test; P<0.05). RT significantly reduced blood glucose (TD = 449 ± 17 vs. SD = 572 ± 18 mg/dL) and attenuated BW loss of diabetic animals. DM reduced renal ACE activity in sedentary and trained groups (SD = 3.72 ± 0.48, TD = 3.85 ± 0.40 vs. SC = 9.2 ± 0.59 nmol/min/mg), while RT reduced enzyme activity only in control group (TC = 5.14 ± 0.26 vs. SC = 9.2 ± 0.59 nmol/min/mg). RT reduced renal ACE2 in the control group compared to the others (TC = 0.05 ± 0.0001 vs. SC = 0.09 ± 0.004, SD = 0.09 ± 0.003, TD = 0.10 ± 0.002 μM/min/mg), with no effect of diabetes on enzyme activity. RT improved renal function, decreasing urinary volume and albuminuria (DT = 4.13 ± 0.84 vs. SD = 11 ± 2.11 mg/24h) in DT group. The results from the present study show that RT is strongly associated with renoprotection in an experimental model of diabetic nephropathy. Moreover, results show that this improvement on renal function is modulated by other pathways apart from ACE and ACE2 converting enzymes. Financial Support: FAPESP, CAPES, CNPq.
Abstract 238: Interleukin 17 Production Induced by Angiotensin Ii Type 2 Receptor Activation in T Cells Drives Flow-mediated Outward Remodeling of Mouse Resistance Arteries
Authors: Caillon Antoine , Grenier Céline , Grimaud Linda , Ngu Yen Phuc Minh Chau , Toutain Bertrand , Loufrani Laurent , Delneste Yves , Henrion Daniel
Published: March 2022

Objective:
Flow (shear stress)-mediated outward remodeling of resistance arteries is involved in collateral growth during postischemic revascularization. Besides the NO pathway the involvement of inflammatory factors and reactive oxygen species has been suggested in this remodeling although the mechanism and the sequence remains unknown. As T cells drive the inflammatory response, we investigated their role in flow-mediated remodeling. Mouse mesenteric resistance arteries (250μm internal diameter) were submitted in vivo to a chronic increase (144±18 to 239±25 μl/min) in blood flow. Arteries were collected for in vitro analysis after 1 to 7 days.

Results:
After 1 week, remodeling occurred in high flow (HF) arteries (diameter increased from 242±21μm to 324±20μm, n=9 per group, P<00.1) in control mice. No remodeling occurred in nude mice and in mice treated with anti-CD3 antibodies. In control arteries lymphocyte T cells accumulate around HF arteries at day 1 (shown using both immunohistology and flow cytometry) and they expressed angiotensin II type 2 receptor (AT2R). In isolated T cells AT2R stimulation induced preferentially IL-17 production (20±6pg/ml to 50±5pg/ml, n=7 per group, P<00.1). We then tested the occurrence of remodeling in mice lacking AT2R or IL-17. In both AT2R-/- and IL-17-/- mice diameter expansion did not occur whereas T cells and myeloid cells accumulation around the HF artery was not affected. IL-17 infusion (5ng/h, Alzet osmotic minipump) to nude mice and to AT2R-/- mice restored HF remodeling to control level. In order to confirm the role of AT2R in flow-mediated remodeling in another vascular territory, neovascularization was examined using an ischemic skin flap model in AT2R-/- mice. As in mesenteric arteries, the absence of AT2R prevented diameter expansion in the arteries feeding the skin flap.

Conclusion:
we demonstrate for the first time the involvement of lymphocytes T cells polarization into TH17 by angiotensin II type 2 receptor in flow-mediated outward remodeling of resistance arteries.
Abstract 239: CD8+ T Cells Elicited by an ApoB-100 Related Peptide Vaccine Mediate Protection Against Angiotension II-induced Hypertension and Renal Fibrosis
Authors: Honjo Tomoyuki , Dimayuga Paul C , Chyu Kuang-Yuh , Zhao Xioaning , Zhou Jianchang , Yano Juliana , Lio Wai Man , Trinidad Portia , Cercek Bojan , Shah Prediman K
Published: March 2022

Background:
T cells play an important role in angiotensin II (AngII)-induced hypertension but the specific subtype involved remains unknown. We recently reported that immunization with apoB-100 related peptide p210 modified CD8+ T cell function, and reduced mean blood pressure (BP) and renal inflammation in AngII-infused apoE (-/-) mice. In this study, we assessed the role of CD8+ T cells in p210 vaccine mediated renal protection.

Method and Results:
ApoE (-/-) mice were immunized with p210/cBSA/Alum (p210; 100 μg) at 7, 10, and 12 weeks of age. Mice receiving PBS or cBSA/Alum (cBSA) served as controls. At 10 weeks of age, mice were subcutaneously implanted with an osmotic pump which released AngII (1 mg/Kg/min), and were euthanized 4 weeks later. Flow cytometric analysis showed p210 treated mice had significantly increased CD8+CD25+IL-12+ splenic T cells indicating CD8+ T cell activation (Table). Renal fibrosis assessed by Masson-Trichrome staining and pro-fibrotic gene expression was significantly reduced in p210 vaccine treated group (Table). P210 immunized mice with CD8+ T cell depletion by a monoclonal antibody (Ab) developed higher mean BP compared to similarly treated mice receiving isotype Ab or no Ab injection ( no Ab injection = 123±1 mmHg, CD8 Ab = 132±1 mmHg*, isotype Ab = 123±2 mmHg, *p<0.05, N=8, 7, 5 respectively, *p<0.05 vs no Ab injection.). Depletion of CD8+ T cells also increased renal fibrotic gene expression compared to no Ab injection (Table).

Conclusion:
CD8+ T cells elicited by an apoB-100 related peptide vaccine mediate protection against AngII-induced hypertension and renal fibrosis.
Abstract 245: Role of Endogenous T Cell Receptors in the Development of Hypertension
Authors: Trott Daniel W , Chen Wei , Harrison David G
Published: March 2022

The purpose of this investigation was to determine if endogenous T cell receptors (TCR) are required for the development of hypertension. Our laboratory has preliminary data suggesting that hypertension is associated with an oligoclonal TCR expansion of CD8+ T cells in mouse kidney. To determine whether mice with an altered TCR phenotype are protected against hypertension, OT1 mice, which express a transgenic TCR specific for ovalbumin on CD8+ cells, and OT1xRag-1-/- mice were infused with angiotensin II (490ng/min/kg) or vehicle. Blood pressure was measured by telemetery. After 2 weeks OT1 mice infused with angiotensin II exhibited increased blood pressure (171 ± 10 systolic, 113 ± 16 diastolic, mean ± S.E.M. mmHg) compared to controls (117 ± 15 systolic, 83 ± 7 diastolic). The hypertensive response was blunted in OT1xRag-1-/- mice that received angiotensin II (142 ± 12 systolic, 85 ± 13 diastolic). Analysis of kidney resident CD8+ TCR by flow cytometery revealed that 6.8 ± 2.9% and 5.2 ± 1.7% in sham and angiotensin II treated OT1 mice respectively were non-transgenic whereas 0.3 ± 0.2% and 0.5 ± 0.1% in sham and angiotensin II treated OT1xRag-1-/- mice respectively were non-transgenic. These data together with the finding that hypertension promotes an oligoclonal TCR expansion in kidney suggest that endogenous TCR are required to respond to neoantigens and mediate the development of overt hypertension.
Abstract 249: Modulation of Angiotensin II-Induced Hypertension by Cytochrome P450 1B1 Metabolites of Estrogen in Female Mice
Authors: Jennings Brett L , Pingili Ajeeth K , Khan Nayaab S , Malik Kafait U
Published: March 2022

Pre-menopausal females are less susceptible to increases in blood pressure (BP) compared to males in various models of experimental hypertension, and this protection is lost following ovariectomy (OVX). We reported that in male rodents, cytochrome P450 (CYP) 1B1, via generation of reactive oxygen species, contributes to angiotensin (Ang) II- and DOCA-salt-induced hypertension and in SHR. Recently it has been shown that dexfenfluramine causes pulmonary arterial hypertension by a CYP1B1-dependent mechanism in female, but not male mice. Whether sex differences exist in the contribution of CYP1B1 to systemic hypertension is not known. Since CYP1B1 has a high affinity for estrogen, and estrogen and its metabolites exert both pro- and anti-proliferative effects, this study was conducted to determine the contribution of CYP1B1, estrogen, and its metabolites in the development of Ang II-induced hypertension in 8 weeks old female

Cyp1b1
+/+

and

Cyp1b1
-/-

mice. Mice were infused with Ang II (700 ng/kg/min) or vehicle with or without 2-hydroxyestradiol (2-OHE), 4-OHE (1.5 mg/kg/day, i.p.), or the CYP1B1 inhibitor, 2,4,3′5′-tetramethoxystilbene (TMS, 300 μg/kg, every 3
rd
day, i.p.), for 2 weeks, and BP was measured twice weekly by tail cuff. Ang II increased BP to a greater degree in

Cyp1b1
-/-

than in

Cyp1b1
+/+

mice (119 ± 3 to 171 ± 11 mmHg vs. 120 ± 4 to 149 ± 4 mmHg, respectively, P < 0.05). The increase in BP produced by Ang II in

Cyp1b1
-/-

mice was reduced by treatment with 2-OHE (171 ± 11 vs. 140 ± 8 mmHg, P < 0.05). However, treatment with 4-OHE in

Cyp1b1
+/+

mice enhanced the hypertensive effect of Ang II (190 ± 4 vs. 149 ± 4 mmHg, P < 0.05), as did TMS (187 ± 5 vs. 149 ± 4 mmHg, P < 0.05). These agents did not affect basal BP in

Cyp1b1
+/+

or

Cyp1b1
-/-

mice. Furthermore, our preliminary data show that OVX increased the hypertensive effect of Ang II after 7 days of infusion in

Cyp1b1
+/+

mice (171 ± 10 vs. 140 ± 4 mmHg, P < 0.05) but not

Cyp1b1
-/-

mice (167 ± 8 vs. 169 ± 7 mmHg, P > 0.05). These data suggest that estrogen metabolites of CYP1B1 exert both anti- (2-OHE) and pro-hypertensive (4-OHE) effects, and that the low pressor effect of Ang II in female mice is most likely mediated by the latter. Moreover, agents that inhibit CYP1B1 activity could promote development of systemic hypertension in female mice.
Abstract 250: 20-HETE is Increased and EETs are Decreased in the Renal Vasculature of a Rat Model of Polycystic Ovary Syndrome
Authors: Moulana Mohadetheh , Roman Richard J , Baker Rodney , Reckelhoff Jane F
Published: March 2022

Polycystic ovary syndrome (PCOS) is the most common hormonal disorder among women during their reproductive years. Enlarged ovaries containing numerous small cysts, elevated circulating androgens, modest increases in blood pressure, and insulin resistance, are among the main symptoms of PCOS. However, the mechanisms responsible for higher blood pressure have not been elucidated. We have characterized a rat model of PCOS, the hyperandrogenemic female (HAF) rat that exhibits characteristics of PCOS in women. The present study was performed to test the hypothesis that 20-HETE and EET levels and expression of the cytochrome P450 (CYP) 4A ω-hydroxylases are different in HAF and placebo rats. Dihydrotestosterone (DHT) (7.5 mg/90 days)

or placebo pellets were implanted subcutaneously in female SD rats (n = 6/group) at 4

weeks of age. Kidneys were removed at 12 weeks of age, preglomerular vessels isolated, and 20-HETE, EETs, and ω-hydroxylase activity were measured by liquid chromatography-mass spectroscopy; CYP 4A protein was measured by western blot. The basal renal microvascular level of 20-HETE (0.15± 0.02 vs. 0.81± 0.08 pmol/mg; p<0.001) and renal vascular ω-hydroxylase activity (0.20± 0.04 vs. 0.69± 0.14 pmol/min/mg; p<0.01) were higher in HAF compared to placebo controls. In addition, CYP4A protein expression was increased by 79% (0.31 vs. 0.40 AU; p<0.005) in HAF rats. In contrast, the basal renal microvascular level of EETs was reduced in HAF rats (40.80± 11.96 vs. 2.33± 1.74 pmol/mg; p<0.005). The data suggest that renal production of 20-HETE is increased in HAF, and may contribute to renal vascoconstriction, whereas EETs are decreased which may contribute to reduced vasodilation of preglomerular vasculature. These data suggest that androgens may affect intrarenal vascular eicosanoids in women with PCOS to mediate their increases in blood pressure.