bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract P071: Mineralocorticoid Receptor Overexpression in Adipose Tissue Leads to Metabolic Syndrome and Induction of Prostaglandin D2 Synthase PTGDS and lipocalin-2 NGAL
Authors: Feraco Alessandra , Urbanet Riccardo , Nguyen Dinh Cat Aurelie , Venteclef Nicolas , El Mogrhabi Soumaya , Rossignol Patrick , Fallo Francesco , Touyz Rhian M , Jaisser Frederic
Published: March 2022

Objectives:
Metabolic Syndrome (MetS) is a cluster of metabolic risk factors associated with a higher risk for cardiovascular events. Adipose tissue (AT) plays a central role in the obesity-related metabolic abnormalities and it’s known that Mineralocorticoid Receptor (MR) activation affects adipocyte differentiation and function. The purposes of this study were to evaluate the molecular and metabolic consequences of adipocyte-targeted increase in MR expression in mice.

Results:
In our study we showed that MR expression is increased in visceral adipose tissue (VAT) in a preclinical mouse model of MetS. Thus, we generated a double transgenic mouse model with a conditional and inducible overexpression of MR in AT (Adipo-MORE), demonstrating that increased expression of MR in AT contributes to MetS development with multiple metabolic abnormalities, including visceral obesity (VAT mass, control-MR 5.0±0.6 adipo-MORE 10.4±1.8, p<0.05), insulin resistance (HOMA index, control-MR 8.9±0.8 adipo-MORE 14.5±2.2, p<0.05) as well as dyslipidemia (total cholesterol, mg/dL control-MR 86.0±6.0 adipo-MR 109±6, p<0.05). We also identified prostaglandin D2 synthase (PTGDS) as a novel mediator of adipogenic effects of MR activation in adipocytes. Since adipokines play pivotal roles in the regulation of insulin sensitivity in obesity, we also focused on the role of lipocalin-2 NGAL, recently identified in our laboratory as a target of aldosterone action in cardiovascular system, as new adipocyte MR target. It’s known that NGALKO mice fed a high fat diet are protected from obesity-induced cardiovascular dysfunctions. In our model we observed an increase of NGAL mRNA levels (3 fold increase, p<0.05) in VAT. Moreover, NGAL was induced (2-8 fold increase, p<0.05) in primary cultures of adipocytes differentiated ex vivo from adipo-MORE mice in presence of aldosterone stimulation compared to controls.

Conclusions:
We demonstrated that PTGDS is a novel direct MR target in rodent adipocytes mediating adipogenic effects of MR activation. We also found that NGAL is induced in presence of MR overexpression in adipocytes in vivo and ex vivo. In consideration of our results, other analyses are necessary to better understand the role of NGAL as adipocyte MR target.
Abstract P074: Role of Peroxisome Proliferator-Activated Receptor-γ (PPAR) in the Pro-opiomelanocortin (POMC) Neuron-Mediated Regulation of Energy Balance
Authors: Stump Madeliene , Guo Deng Fu , Rahmouni Kamal , Sigmund Curt D
Published: March 2022

PPARγ, a master regulator of adipogenesis, was recently shown to affect energy homeostasis through its actions in the brain. Deletion of PPARγ in mouse brain, and specifically in the POMC neurons, results in resistance to diet-induced obesity. To study the mechanisms by which PPARγ in POMC neurons controls energy balance, we generated a transgenic mouse model in which a dominant-negative mutant (P467L) form of PPARγ is conditionally expressed in POMC neurons. The transgene will express both human PPARγ-P467L and the tdTomato reporter gene after it is selectively activated by a Cre-recombinase driver. Here, POMC-Cre was used to direct POMC neuron-specific expression of the transgene. Co-localization of tdTomato and ACTH staining in the Arcuate nucleus of POMC-Cre X PPARγ-P467L double transgenic (DTg) mice demonstrates successful transgene activation in POMC neurons. Interestingly, 25 week treatment with 60% high fat diet resulted in no difference in body weight in DTg mice compared to littermate controls (55.7±1.0 g vs 57.1±1.2 g, n>11). However, feeding these mice a 10% fat isocaloric-matched control diet led to a significant increase in the body weight of DTg mice compared to littermate controls (36.0±0.9g vs. 31.7±0.5g, p<0.0001, n=10). The increased body weight in DTg was due to increased adipose mass (DTg: 23.9±2.4% vs. control: 16.8±2.2%, p=0.04, n=10) and decreased lean mass (DTg: 60.5±2.0% vs. control: 66.5±1.7%, p=0.04) respectively, as demonstrated by nuclear magnetic resonance spectroscopy. We observed no difference between genotypes in food intake (DTg: 2.1±0.1g vs. controls: 2.2±0.1g, n>9) and cumulative body weight change (DTg: -4.5±0.7% vs. controls: -4.4±1.0%, n>9) in response to 1mg/kg (i.p.) leptin. Since interference with PPARγ in POMC neurons does not impede sensitivity to leptin, we speculate that increased weight gain in DTg mice on low fat diet may be due to differences in food intake, decreased sympathetic drive to adipose tissues, and/or decreased energy expenditure, mechanisms which are currently under investigation. We conclude that PPARγ in POMC neurons may play a role in energy balance under certain dietary conditions.
Abstract P078: Cardiovascular Autonomic Impairment Triggers Cardiometabolic Dysfunction in an Experimental of Metabolic Syndrome
Authors: Bernardes Nathalia , Conti Filipe F , Dias Danielle d , Brito Janaina d , Malfitano Christiane , Irigoyen Maria-Cláudia , De Angelis Kátia
Published: March 2022

The aim was evaluate on time-course of changes in cardiometabolic dysfunction model. Male Wistar and SHR rats were divided: Control (C), hypertensive (H), hypertensive + fructose (HF). The fructose overload (100g/L) was initiated at 30 days (d) of life. All evaluations were at 37, 45, 60 and 90 d of age. Arterial pressure (AP) signals were directly recorded. The baroreflex sensitivity was evaluated by the tachycardic (TR) and bradycardic (BR) responses to AP changes. Cardiovascular autonomic modulation was evaluated by spectral analysis. Inflammatory markers were evaluated in adipose tissue. No differences were observed in glycemia and triglycerides between the groups. The SHR groups showed increased systolic (SAP), diastolic (DAP) and mean arterial pressure (MAP) compared to control groups in all times of evaluation. Additionally, SHR groups progressively increased AP during the time course evaluation. The HF group showed additionally increased in DAP and MAP (90: MAP: C: 114±1, H: 165±3, HF: 179±3 mmHg) vs. H group. The cardiac parasympathetic modulation (HF-IP) was decreased H group in 37 d and HF group in 37, 90 d vs. C group. The VAR-PAS was higher in SHR groups vs. C group in all times evaluated. The HF group showed additionally increase in VAR-PAS during the time course evaluation (45, 90 vs. 37 d). The vascular sympathetic modulation (LF-PAS) was higher H group in 60, 90 d vs. C group. The HF group showed increased LF-PAS in 45, 60, 90 d vs. C group, and vs. 37 d in the same group; and showed additionally increased in 90 d vs. H group (90: C: 3.7±0.6 H: 6.2±0.9 HF: 8.7±0.5 mmHg2). The spontaneous baroreflex sensitivity was lower in SHR groups all the times evaluated (90: C: 0.80±0.1 H: 0.51±0.05 HF: 0.32±0.02 ms/mmHg). The H group showed decreased BR in 60, 90 d vs. C group, the HF group showed decreased in 37, 45, 60, 90 d vs. C group (90: C: 1.15±0.1 H: 0.80±0.05 HF: 0.77±0.07 bpm/mmHg). The HF group showed decreased TR in 60, 90 d vs. C group. Increased TNF alpha and IL-6 were observed only in 45 and 60 d in HF group. The results showed that cardiometabolic dysfunctions are time-dependent in SHR, and fructose overload induces additional dysfunctions in this model. Furthermore, the baroreflex impairment seems to precede hemodynamic, metabolic and inflammatory changes in this model.
Abstract P080: New Strategy, Old Remedy: Elucidating the Effect of Hibiscus Sabdariffa in a Neuronal Model of Oxidative Stress
Authors: Shalgum Aiman , Jones Parquita , Angajala Anusha , Glenn Johnny , Dobbs Jamel , Bowers Pecoria , Griffin Gerald , Collier Willard , Dhanasekaran Muralikrishnan , Bradford Chastity N
Published: March 2022

Introduction:
Neurogenic hypertension is characterized by increased blood pressure, neuroinflammation, and neuronal oxidative stress. Thus, its pathophysiology dictates a unique therapeutic strategy. It is established that Hibiscus sabdariffa (HS) plays a beneficial role in decreasing blood pressure, yet the role of HS as an anti-oxidant defense system in neurogenic hypertension has not been investigated. Our objective was to generate an in vitro model of neuronal oxidative stress to test the hypothesis that HS creates a neuroprotective antioxidant defense system.

Methods:
To create a model of oxidative stress, SY5Y, neuroblastoma cells, were treated with 32uM hydrogen peroxide. To determine the effect of methanol-extracted HSE (50 and 100ug n ≥ 8) in this model of oxidative stress, SY5Y cells were treated with vehicle-PBS, HSE (24hrs), pretreated with HSE (24hrs) and then treated with H2O2 (8hrs). Using molecular techniques markers of oxidative stress were measured and percentages were out of 100. Data was normalized to the control and analyzed by one way ANOVA (p value < .05 was considered significant).

Results:
HSE significantly increases cell viability (50 or 100ug > 99%) compared to H2O2 alone (43%). HSE significantly decreases ROS generation (113%-50ug versus 89%-100ug) compared to H2O2 (120%). HSE significantly decreases lipid peroxidation (11.3%-50ug versus 10.2%-100ug) compared to H2O2 (135%). HSE significantly increases GSH content (72%-50ug versus 91%-100ug) compared to H2O2 (58%). HSE significantly increases catalase activity (31%-50ug versus 51%,-100ug) compared to H2O2 (23%). HSE significantly increases mitochondrial complex 1 activity (125%-50ug versus 229%-100ug) compared to H2O2 (26.2%). There was no significant change in superoxide dismutase activity amongst groups.

Conclusion:
These observations suggest that HSE creates an antioxidant defense system that provides cytoprotection against H2O2-induced neuronal oxidative stress. Thus, HS, a known anti-hypertensive should be revisited to investigate its in vivo role in neurogenic hypertension.
Abstract P085: Angiotensin II Type 2-receptor Agonist C21 Reduces Proteinuria and Oxidative Stress in Kidney of High-salt Fed Obese Zucker Rats
Authors: Patel Sanket N , Hussain Tahir
Published: March 2022

Oxidative and nitrosative stress have been implicated in hypertension and kidney diseases. Recently we reported blood pressure reducing effect of AT2-receptor agonist C21 in high sodium-diet (HSD)-fed obese Zucker rats. In the present study we investigated C21-mediated reno-protection against HSD-related oxidative and nitrosative stress in obese Zucker rats. Obese rats (13 week) were fed normal sodium-diet (NSD) or HSD 4%, for 14 days, with/without C21, delivered subcutaneously via osmotic pump, 1 mg/kg/day. Compared with NSD controls, HSD-fed rats exhibited reduced eGFR (15.2±2.3 vs. 43.5±9.9 μL/min) and increased proteinuria (0.18±0.02 vs. 0.12±0.02 μg/min) and albuminuria (0.19±0.03 vs. 0.12±0.02 μg/min), which were associated with decreased cortical expression of the protein recycling receptors, megalin (0.25±0.02 vs. 0.47±0.07) and cubilin (0.77±0.16 vs. 1.27±0.14). C21 improved eGFR (27.4±4.9 μL/min) and reduced proteinuria (0.11±0.02 μg/min) and albuminuria (0.08±0.01 μg/min) without any effects on megalin or cubilin. Cortical NADPH oxidase (NOX) activity in HSD-fed rats was increased by 3-fold (0.37±0.07 vs. 0.13±0.04 ΔRLU/μg protein/min), while enzymatic defense (catalase/superoxide dismutase activity ratio) was unchanged. Urinary excretion of H2O2 (41.7±4.7 vs. 23.0±2.1 μM/mg creatinine) and 8-isoprostanes (7.9±1.6 vs. 1.4±0.2 pg/μg creatinine), indices of oxidative stress, were elevated in HSD-fed rats. C21 reduced NOX activity (0.11±0.04 ΔRLU/μg protein/min) as well as the excretion of H2O2 (30.5±3.0 μM/mg creatinine) and 8-Isoprostanes (4.1±1.0 pg/μg creatinine) in HSD-fed rats. Similarly, C21 also reduced carbonyls (5.5±1.4 vs. 10.5±0.9 μM/mg protein), an indicator of protein modification caused by oxidative stress. 3-nitrotyrosine, an indicator of nitrosative stress, remained unaltered in HSD and/or C21 groups. C21-treatment of HSD-fed rats improved plasma nitrites (22.1±2.2 vs. 15.3±0.5 μM/L), which could have been due to reduced oxidative stress in this group. Together results indicate that AT2R agonist exerts antioxidant effects, possibly by reducing NOX activity and by rescuing NO levels, which potentially contributes to the reduction in salt-induced proteinuria/nephropathy in obesity.
Abstract P098: G-protein Coupled Estrogen Receptor Stimulates Capillary Formation by Human Umbilical Vein Endothelial Cells via ALK1-SMAD 1/5/8 Pathway Activation
Authors: Unterleutner Elisabeth , Rigassi Lisa , Barchiesi Federica , Imthurn Bruno , Dubey Raghvendra K
Published: March 2022

Estradiol (E2) induces vascular repair by promoting endothelial growth and capillary formation. Based on our previous findings that the capillary stimulating effects of E2 are mimicked by its non-permeable analog, BSA-tagged E2, we hypothesize that the stimulatory effects are potentially mediated via the newly discovered membrane bound G-protein coupled estrogen receptor (GPER). To investigate this, we assessed capillary formation by endothelial cells in response to E2, and GPER agonist (G1)and antagonist (G15) in a 2-D matrigel based assay. Capillary formation was assessed microscopically by quantifying junction/sprout formation and capillary length. E2 (10nM) increased capillary formation and this effect was mimicked by G1 (10nM; stimulated from 100% to 517±46% and 210±41%, respectively; p<.05 vs control). The effects of E2 and G1 were significantly abrogated by G15 (100nM; p<.05), suggesting a role for GPER in mediating the capillary stimulatory effects of E2. Because G-protein coupled mechanisms, Akt, ALK1 and SMAD1/5/8 are involved in capillary formation, we investigated their role in GPER induced capillary formation. ECs expressed GPER, ALK1 and SMAD1/5/8. Treatment with G1 (10-100nM) up-regulated the expression of ALK1 from 100% to 168±21% (p<.05 vs control) and phosphorylated SMAD1/5/8 from 100% to 208±36% (p<.05 vs control). Similar to capillary formation the stimulatory effects of G1 on SMAD1/5/8 were blocked significantly (p<.05) by G15 (100nM), Pertussis Toxin (0,1ng/ml; G protein pathway inhibitor), LY294002 (5μM; Akt/PI3K inhibitor) and ALK1Fc (100ng/ml; specific ALK-1 antagonist). Silencing of GPER and SMAD1 by siRNA (50nM) abrogated the stimulatory effects of E2 and G1 on capillary formation, and SMAD1/5/8 and Akt phosphorylation. Moreover, treatment with G1(100nM) upregulated ID-1 expression from 100% to 179±26% (p<.05 vs control), a downstream target of ALK1/pSMAD1/5/8. We conclude that E2 via GPER promotes EC-mediated capillary formation by a mechanism that involves non-genomic activation of ALK1→ pSMAD1/5/8 ↔ pAkt → ID-1. GPER agonists in general may promote healing of injured vascular beds by promoting EC activity leading to more rapid endothelial recovery and capillary formation following injury.
Abstract P101: The Role of Primary Cilia in Vascular Endothelial Cells
Authors: Saternos Hannah , Hossain Saad Zubayer , AbouAlaiwi Wissam
Published: March 2022

Primary cilia are mechanosensory organelles that are projected into the lumen of blood vessels. It has been demonstrated that vascular endothelia require primary cilia to sense and transmit external mechanical stimuli into internal biochemical reactions. One of these reactions includes the biosynthesis and release of nitric oxide, which is one of the most potent endogenous vasodilators. This idea has
only
been investigated in cultured endothelial cells
in vitro
. Based on this finding, however, a very bold hypothesis is formed to test that abnormal cilia function results in vascular hypertension. Our laboratory has recently generated and obtained several conditional mouse models to specifically study the function and structure of primary cilia in vascular endothelia. These models include
1)
mice without cilia function (
Pkd1
or
Pkd2
);
2)
mice without cilia structure (
Tg737
or
Kif3a
). Our data indicate that mice with abnormal cilia function (
Pkd1
) or structure (
Tg737
) show significantly higher systolic (150±19 for

Pdgfbcre:Pkd1
flox/flox

and 147±10 for

Tie2Cre:Tg737
flox/flox

vs. 128±9 for wild-type) and diastolic (120±21 for

Pdgfbcre:Pkd1
flox/flox

and 120±11 for

Tie2Cre:Tg737
flox/flox

vs. 102±7 for wild-type) blood pressure than the corresponding wild-type mice. Because there is a positive and continuous correlation between blood pressure and cardiovascular diseases, satellite hypotheses are developed to look at the pathophysiological roles of endothelial cilia in cardiac functions and focal vascular diseases
in vivo
. Our data clearly point towards deteriorating phenotypes in the cardiac muscle, including cardiac fibrosis due to an increased cardiac workload. As a result, a heart-to-body weight ratio was significantly increased by 17 weeks old (0.008

PdgfbCre;Pkd1
f/f

vs. 0.006

Pkd1
f/f

).The present study will very likely provide new insights for hypertension and offer advanced scientific understanding of vascular endothelial cilia in other cardiovascular diseases.
Abstract P107: Metabolic Syndrome and Human AT1R Expression in Transgenic Mice: Implications of Haplotype-dependent Transcriptional Regulation
Authors: Rana Anita , Jain Sudhir , Eren Deniz , Sirianni Natalie , Kaw Meenakshi , Kumar Ashok
Published: March 2022

Angiotensin II (Ang II) contributes to the pathophysiologies of cardiovascular and renal systems. Angiotensin receptor type 1 (AT1R) mediates these effects, and genetic variations that increase AT1R can increase these pathological outcomes. Physiological variables like age or pathologies like the metabolic syndrome alter the transcriptional milieu of cells and can provide for feedback activation of genes. In this regard, we have identified two haplotype blocks of single nucleotide polymorphisms (SNPs) in the hAT1R gene: haplotype II (Hap II: -810A, -713G, -214C, -153G) and I (Hap I: -810T, -713T, -214A, -153A). In clinical studies, Hap I is linked to human hypertension. We have generated transgenic mice (TG) with haplotypes-II and I of the hAT1R gene to study its regulation during metabolic syndrome (MetS). At baseline, ChIP assay shows increased RNA-Pol II binding (~1.6 fold higher) to the chromatin extracts from renal tissues of adult (4-6 months) male Hap I-TG mice with increased hAT1R expression (~6 fold higher). This was accompanied by higher baseline blood pressure in Hap I-TG mice (Hap I- 126±3 vs. Hap II- 115±4, p<0.05). To induce MetS, these mice were fed Western diet (45% Kcal from fat and carbohydrate each) for 12 wks. Change in body weight is higher (p<0.05) in Hap I (161.5 gm.) vs. Hap II (10.41.2 gm.) mice. MetS phenotype is characterized by increase in blood pressure that is significantly greater in Hap I mice (1362 vs. 1203 in Hap II). Transcription factors, p38/MAPK and STAT3, were induced by MetS to similar extent in both groups. However, MetS-induced up regulation of the hAT1R gene is significantly higher in vascular tissues of Hap I mice (≈6 fold), when compared to Hap II. Complementary experiments show increased inflammatory and redox markers in vascular tissues of Hap I mice, when compared to Hap II, during MetS; including, IL1 (2.2 fold), IL6 (1.8 fold), NOX1 (3.5 fold), VEGF (1.5 fold), and ICAM1 (14 fold). Thus, haplotype-dependent transcriptional regulation of the hAT1R gene causes increased hAT1R expression and blood pressure, in Hap I TG mice. Importantly, MetS exacerbates this differential gene-expression regulation, further increasing hAT1R and promoting a prooxidant/inflammatory milieu in mice with Hap I.
Abstract P115: Effects of Exercise Training on Renal Damage and Renin-angiotensin System in Dahl Salt-sensitive Rats
Authors: Sakuyama Akihiro , Ito Osamu , Ogawa Yoshiko , Komatsu Miwa , Hu Gaizun , Miura Takahiro , Muroya Yoshikazu , Suda Chihiro , Kohzuki Masahiro
Published: March 2022

Exercise training (Ex) has anti-hypertensive and renal protective effects. Renin-angiotensin system (RAS) are involved in the regulation of blood pressure and renal damage. In this study, we investigated the effects of the Ex on renal RAS in Dahl salt-sensitive rats. Six-week-old, male Dahl salt-sensitive rats were divided into four groups: 1) normal salt diet (NS); 2) NS + Ex; 3) high salt diet (HS); 4) HS+ Ex. NS or HS groups were fed diet containing 0.6% or 8% NaCl, and treadmill running was performed in Ex groups for 8 weeks (5 days/week; 60 min/day at 16-20 m/min, 0% grade). Systolic blood pressure (SBP) was monitored by tail-cuff method. Urine samples were collected on ice for 24 hours with metabolic cage. After 8 weeks, protein expression of RAS components in renal cortex (CO) and outer medulla (OM) were investigated by Western blotting. HS significantly elevated SBP (209±6 vs. 114±3 mmHg), and Ex did not change SBP (205±8 mmHg). HS significantly decreased creatinine clearance (0.96±0.04 vs. 2.57±0.07 ml/min/g kidney weight), but Ex significantly mitigated creatinine clearance (1.35±0.18 ml/min/g kidney weight). HS significantly increased urinary protein excretion (433±37 vs. 16±2 mg/day), but Ex significantly suppressed urinary protein excretion (327±22 mg/day). HS induced glomerular sclerosis, but Ex suppressed it. HS increased angiotensinogen expression (138 % and 328 %) and decreased renin expression (47% and 24%) in the CO and OM. HS increased angiotensin II type 1 (AT1) receptor expression in the OM (149%) and Mas receptor expression in the CO (198%), but decreased angiotensin II type 2 (AT2) receptor expression in the CO and OM (53% and 36%) and Mas receptor expression in the OM (20%). Ex improved HS-increased angiotensinogen and AT1 receptor expressions only in the OM. Ex improved HS-decreased renin expression in the CO and OM, and HS-decreased AT2 and Mas receptor expressions only in the OM. These results indicated that Ex improves HS-induced renal damage independently of SBP with specific changes of RAS components in the OM. Ex may have beneficial effects in HS-induced renal damage.
Abstract P117: Lysine Specific Demethylase-1 Deficiency Accelerates the Development of Renal Damage and Hypertension During Long Term Exposure to Sodium
Authors: Katayama Rangel Isis A , Chong Cherish , Williams Gordon H , Pojoga Luminita H
Published: March 2022

Long term exposure to salt is a demonstrated risk factor for hypertension (HT), as well as cardiovascular and renal (CVR) outcomes. We recently proposed a novel contributor to the etiology of salt-mediated HT: the Lysine Specific Demethylase-1 (LSD1). We showed that LSD1 deficiency (in mice) and LSD1 gene variants (in humans) associate - in response to
short term
(one week) of sodium loading - with dysregulated renal sodium handling, volume expansion and HT, and RAAS dysfunction. However, the timeline and severity of these changes during
long term
exposure to sodium, and the protective effects of sodium restriction in LSD1 deficient states have yet to be determined. This study aimed (1) to evaluate the timing of onset for changes in CVR health during long term sodium loading in LSD1 deficient mice and (2) to assess whether a low salt (LS) diet can prevent these effects. LSD1 heterozygous (HET) and WT mice were randomized to high salt (HS) or LS and followed longitudinally for 6 months. BP, plasma aldosterone (Aldo) and albumin/creatinine ratios (A/C) were assessed monthly.

The SBP (mm Hg) increased progressively during the study, and reached significance on the 5
th
and 6
th
month for HS-HET and HS-WT (141±4 and 134±3, respectively, both p<0.05 vs. baseline) but not for LS-HET and LS-WT (128±6 and 120±6, respectively). The SBP effects were driven by a significant interaction between genotype and age (p<0.05). Similar results were obtained for DBP (p<0.05), suggesting a volume mediated effect. HS plasma Aldo was appropriately suppressed.

The A/C (μg/mg) was progressively increased in both HS groups, one month prior to the BP change. Namely, A/C reached significance on the 4
th
and 5
th
month for HS-HET and HS-WT (46±4 and 48±9, respectively, both p<0.05 vs. baseline). The LS diet prevented these changes in both genotypes.

Our novel study shows that long term exposure to HS induces kidney damage followed by BP increase, and that these changes are initiated earlier in the LSD1 HET, suggesting LSD1 as a critical component of mechanisms involved in CVR health. Moreover, long term sodium restriction prevented the development of both target organ damage and HT in this model, suggesting that this dietary intervention may be particularly efficient in human carriers of LSD1 gene variants.
Abstract P118: Differential Effects of Renal α- and β-Adrenoreceptors in the Development of Sodium Chloride Cotransporter (NCC) Dysfunction and Salt-sensitive Hypertension
Authors: Walsh Kathryn R , Wainford Richard D
Published: March 2022

Aim:
Determine the contribution of α vs. β-adrenoeceptors in NCC dysfunction & the development of salt-sensitive hypertension (HTN).

Methods:
Sprague-Dawley rats receiving a sc saline, NE (600ng/min), NE with propranolol (Pro; 9.9mg/kg/day) or NE with prazosin (Praz; 2.5mg/kg/day, orally) infusion were fed a 0.4% (NS) or 8% NaCl (HS) diet for 14 days (N=6). On day 14, MAP & NCC activity (peak natriuresis to iv hydrochlorothiazide infusion; HCTZ; 2mg/kg) was assessed. Immunoblotting for NCC, STE-20 Alanine/Proline kinase (SPAK), & oxidative stress response-1 (OxSR1) was performed on kidney cortex tissue.

Results:
NE infusion results in HTN that is exacerbated by HS (MAP [mmHg] NE+NS 151±3; NE+HS 171±4; p<0.05). Excess NE prevents HS-evoked suppression of NCC function and NCC & SPAK expression. NE:Pro co-infusion significantly decreased MAP (MAP NE:Pro+NS 136±4; NE:Pro+HS 136±4), regardless of Na+ intake, when compared to rats receiving the NE infusion, but failed to restore HS-evoked suppression of NCC activity (similar to results observed when AT1Rs are chronically antagonized during NE infusion; data not shown), or NCC, SPAK, or OxSR1 expression despite reduced baseline SPAK & OxSR1 compared to NE alone. NE:Praz co-administration abolished the salt-sensitive component of NE-induced HTN (MAP NE:Praz+NS 159±5; NE:Praz+HS 156±5) and restored HS-evoked suppression of NCC function and expression (Peak ΔUNaV [μeq/μl] NE:Praz+NS 10.7±1.2; NE:Praz+HS 6.14±1.2; p<0.05). NE+Praz co-administration resulted in HS-evoked suppression of SPAK, but not OxSR1 expression, in addition to decreased baseline expression of SPAK/OxSR1.

Discussion:
Our data suggests that α, but not β, adrenoreceptors are predominantly responsible for NE-mediated NCC dysregulation and the development of salt-sensitive HTN. During a HS intake, α-adrenoreceptors mediate NE-evoked salt-sensitivity via a SPAK-NCC dependent pathway. Chronic antagonism of β-adrenoreceptors decreased MAP independent of direct effects on NCC possibly via actions on the RAS. Our data shows a synergistic relationship between α-/β-adrenoreceptors to upregulate SPAK/OxSR1 during chronic adrenergic stimulation with SPAK as the primary driver of NCC dysfunction & salt-sensitivity.
Abstract P123: Retinol-Binding Protein 7 (RBP7) Regulates Endothelial Function via an Adiponectin-Dependent Mechanism in Angiotensin II-Infused Mice
Authors: Hu Chunyan , Keen Henry L , Vogel Silke , Faraci Frank M , Sigmund Curt D
Published: March 2022

The transcription factor peroxisome proliferator-activated receptor gamma (PPARγ) regulates vascular function and protects against endothelial dysfunction. We reported that mice expressing dominant-negative PPARγ in endothelium exhibit an enhanced pressor response to angiotensin II (Ang II). Given that RBP7 is a PPARγ target expressed in endothelium, we hypothesized that loss of RBP7 would augment endothelial dysfunction caused by Ang II. Using osmotic minipumps, RBP7 knockout (KO) mice and wild type (WT) littermates were infused with subpressor Ang II (120 ng/kg/min, 2 weeks) or saline. Systolic blood pressure (by tail cuff) was not different between KO and WT mice and was not altered by infusion of saline or Ang II. Acetylcholine (ACh, 30 μM) mediated relaxation of carotid arteries was not different between groups during saline infusion, but was selectively impaired in KO mice during Ang II (44±5% in KO vs 71±3% in WT, p<0.05). Aortic superoxide, assessed by hydroethidine staining, was higher after Ang II in KO compared with WT mice. Pre-incubation of carotid arteries with a superoxide scavenger Tempol (1 mM, 30 min) restored ACh-induced relaxation in Ang II-infused KO mice (74±6% in Tempol-KO vs 46±8% in KO). To identify molecular mechanisms, RNA Sequencing was performed using carotid arteries from WT and KO mice fed high fat diet (HFD) or normal chow for 8 weeks. Adiponectin (Adipo), a known PPARγ target gene, was increased ~6-fold in HFD-fed WT mice, a response that was markedly blunted in KO mice. Immunohistochemistry revealed that Adipo was expressed specifically in endothelial cells (co-immunostaining with endothelial specific CD31). Rosiglitazone, a PPARγ ligand, increased carotid artery Adipo protein in WT, but not KO mice. To test the importance of Adipo, carotid arteries from Ang II-infused WT and KO mice were incubated with vehicle or full-length Adipo (5 μg/mL) for 12 hrs. Adipo incubation ameliorated Ang II-induced endothelial dysfunction in KO mice (ACh, 30 μM: 86±3% Adipo-KO vs 58±8% KO) while having no effect in WT mice (ACh, 30 μM: 83±1% Adipo-WT vs 87±3% WT). In conclusion, RBP7 protects the endothelium from oxidative stress-induced dysfunction caused by Ang II. Our data suggest an important role for Adipo in mediating this protective response.
Abstract P125: Visceral Adipose Tissue-derived Serine Protease Inhibitor Prevents the Development of Hypertension Through the Inhibition of Vascular Remodeling via Anti-Oxidative Mechanisms in Spontaneously Hypertensive Rats
Authors: Kameshima Satoshi , Sakamoto Yuzaburo , Okada Muneyoshi , Yamawaki Hideyuki
Published: March 2022

Visceral adipose tissue-derived serine protease inhibitor (vaspin) is an adipokine originally identified in visceral adipose tissue of obese type 2 diabetic Otsuka Long-Evans Tokushima Fatty rats. We have been focusing on the direct vascular effects of vaspin and found that vaspin inhibits: 1) tumor necrosis factor-α-induced inflammatory responses in vascular smooth muscle cells (SMCs) via the anti-oxidative mechanisms, 2) platelet derived growth factor-BB-induced migration of SMCs, 3) apoptosis of vascular endothelial cells (ECs) mediated by methylglyoxal, a metabolite of glucose. Since vascular remodeling via inflammation and migration of vascular SMCs as well as apoptosis of ECs is an important process for the development of hypertension, it is suggested that vaspin has preventive roles on the pathogenesis of hypertension. However, it is not revealed whether vaspin affects the development of hypertension in
in vivo
hypertensive model. The aim of the present study was to explore it. Five-week-old male spontaneously hypertensive rats (SHR) were received an intraperitoneal injection of vaspin (1 μg/kg) or saline once daily for four weeks. Age-matched male Wistar-Kyoto (WKY) rats were used as a control. Blood pressure (BP) was measured using a tail-cuff method weekly. Isolated superior mesenteric arteries were used for the examination of vascular structural changes by a hematoxylin and eosin staining. Reactive oxygen species (ROS) generation was examined by an immunohistochemical staining to 4-hydroxy-2-nonenal (4-HNE), an end-product of lipid peroxidation by ROS. Vaspin significantly inhibited age-dependent elevation of BP in SHR (from 165.5 ± 3.9 mmHg to 153.1 ± 2.5 mmHg, n = 3, P < 0.05). Vaspin significantly inhibited vascular wall hypertrophy in SHR mesenteric artery (from 1.35±0.05 to 0.98±0.08-fold relative to WKY, n = 3, P < 0.05). Moreover, vaspin inhibited an increase of 4-HNE-positive area to vessel area ratio in SHR mesenteric artery (from 13.1±4.4 % to 4.6±0.4 %, n = 3). The present results demonstrate that vaspin inhibits the increase of BP through inhibiting vascular remodeling via anti-oxidative mechanisms in SHR.
Abstract P128: RhoBTB1, a Novel PPARγ Target Gene Regulates Vascular Function
Authors: Mukohda Masashi , Ibeawuchi Stella-Rita C , Hu Chunyan , Quelle Frederick W , Sigmund Curt D
Published: March 2022

Peroxisome proliferator-activated receptor gamma (PPARγ) is a ligand activated transcription factor regulating metabolic and vascular function. We previously reported that mice (S-DN) expressing a dominant-negative PPARγ mutation in smooth muscle cells (SMC) are hypertensive, exhibit impaired vascular relaxation and enhanced contraction, and display reduced expression of a novel PPARγ target gene, RhoBTB1. We hypothesized that RhoBTB1 may play a role in the PPARγ-mediated regulation of vascular function that is disrupted in S-DN mice. To test this, we generated transgenic mice (R+) with tamoxifen-inducible, Cre-dependent expression of RhoBTB1 in SMC. These mice were crossed with S-DN to produce mice (S-DN/R+) in which tamoxifen-treatment (75 mg/kg, ip, for 5 days) increased RhoBTB1 RNA expression in aorta from the reduced level seen in S-DN mice, and restored it to the level of non-transgenic mice. Thoracic aorta from S-DN showed impaired acetylcholine (ACh)-induced endothelial-dependent relaxation, which was reversed by replacement of RhoBTB1 in SMC (43.3±4.4 vs 74.2±1.1 %, p<0.01, n=6). A similar improvement was observed in basilar artery (19.9±6.7 vs 48.1±12.3 %, p<0.05, n=6). Aorta from S-DN mice also displayed severely decreased NO donor (sodium nitroprusside, SNP)-induced endothelial-independent relaxation with a right-shifted SNP dose-response, which was also reversed in aorta from tamoxifen-treated S-DN/R+ mice (p<0.01, n=6). To confirm that these effects were specifically due to replacement of RhoBTB1, we assessed vascular function in tamoxifen-treated S-DN mice. Notably, tamoxifen itself did not affect relaxation in response to ACh or SNP, or contraction in response to KCl, endothelin-1 (ET-1) or Prostaglandin F2α in aorta or basilar artery from S-DN (n=4). Interestingly, contraction induced by ET-1, but not KCl, was enhanced in S-DN aorta, and was not improved by restoring RhoBTB1 expression (n=6). This suggests that RhoBTB1 may function specifically by regulating vasodilation pathways. We conclude that RhoBTB1 plays an important role in facilitating vasodilatation in aorta and basilar artery, and loss of RhoBTB1 function explains the vascular dysfunction observed in response to interference with PPARγ in smooth muscle.
Abstract P130: The Influence of Maternal Obesity on Perivascular Adipose Tissue Function in Offspring
Authors: Zaborska Karolina E , Austin Clare , Wareing Mark , Edwards Gillian
Published: March 2022

Objective:
Maternal obesity pre-programs offspring to develop obesity and associated cardiovascular disease although the underlying mechanism is currently unknown. This study investigated the effect of maternal obesity on perivascular adipose tissue (PVAT) regulation of resistance artery tone.

Design and method:
8 week old female SD rats were fed a 10% fat diet (controls) or 45% fat obesogenic diet (HFD) for 12 weeks before mating, then continued on their respective diets during pregnancy and lactation. Male offspring were provided with 10% fat diet until sacrifice at 12 weeks old. PVAT-intact or -denuded mesenteric arteries (250-300μm internal diameter) from offspring were mounted on a wire myograph. Cumulative concentration-response curves were constructed to the thromboxane A2 receptor agonist U46619 (10nM-3μM) ± 10μM A769662, an activator of AMP-activated kinase (AMPK) or 100μM L-NMMA, a nitric oxide synthase (NOS) inhibitor.

Results:
Body weight, systolic and diastolic blood pressure were significantly increased in HFD dams compared to age-matched controls (391±11 vs 348±12 g; 154±1 vs 145±1 mmHg; 116±1 vs 103±103 mmHg respectively) but no differences were observed between offspring. However, fat pads and insulin levels were increased in both HFD dams versus controls (13.4±1.6 vs 6.5±0.8 g; 1.18±0.09 vs 0.76±0.14 nmol/L respectively) and their offspring (7.2±0.3 vs 6.0±0.2 g; 0.56±0.15 vs 0.17±0.05 nmol/L respectively) versus controls. PVAT exerted an anti-contractile effect in artery segments from offspring of control dams (p<0.001), an effect which was lost in offspring of HFD dams. AMPK activation decreased contractility of both PVAT-denuded and -intact arteries from control offspring (p<0.01; p<0.0001 respectively, n=8); this effect was abolished in PVAT-intact vessels of offspring of HFD dams. Inhibition of NOS increased contractility of both PVAT-denuded and -intact arteries from control offspring (p<0.0001 and p<0.0001 respectively, n=8), revealing a contractile effect of PVAT in control offspring (p<0.01, n=8) but not in arteries from offspring of HFD dams.

Conclusions:
In summary, the diminished anti-contractile effects of PVAT in 12 week old offspring of HFD dams may be due to reduced AMPK and nitric oxide activity.
Abstract P131: Mef2C-MYOCD and Leiomodin1 Suppression by miRNA-214 Promotes Smooth Muscle Cell Phenotype Switching in Pulmonary Arterial Hypertension
Authors: Sahoo Sanghamitra , Meijles Daniel , Al Ghouleh Imad , Tandon Manuj , Cifuentes-Pagano Eugenia , Sembrant John , Rojas Mauricio , Goncharova Elena , Pagano Patrick
Published: March 2022

Background:
In pulmonary arterial hypertension (PAH), smooth muscle cell (SMC) phenotype switching from a terminally differentiated contractile to synthetic state is gaining traction as our understanding of disease progression improves. While maintenance of SMC contractile phenotype is reportedly orchestrated by a MEF2C- Myocardin (MYOCD) interplay, little is known regarding molecular control at this nexus. Moreover, the burgeoning interest in microRNAs (miRs) provides a basis for exploring their modulation of MEF2C-MYOCD signaling, and, in turn, a pro-proliferative, synthetic SMC phenotype. We hypothesized that suppression of SMC contractile phenotype in pulmonary hypertension is mediated by miR-214 via repression of the MEF2C-MYOCD-leiomodin1 signaling axis.

Methods and Results:
In SMCs isolated from a PAH patient cohort and commercially obtained hPASMCs exposed to hypoxia, miR-214 expression was upregulated approx. ~1.5 fold compared to controls (p<0.05). These increases in miR-214 were paralleled by downregulation of MEF2C, MYOCD and SMC-specific contractile proteins, leiomodin1 and smoothelin. Of these, leiomodin1 was directly targeted by the miR. MicroRNA-214 overexpression mimicked the PAH profile, downregulating MEF2C (1±0.054 vs 0.696±0.026, p<0.05) and leiomodin1 (1± 0.051 vs 0.281±0.095, for scrambled control vs miR-214 mimic, p<0.05). Hypoxia significantly reduced expression of SMC-specific contractile proteins, leiomodin1 and calponin1 (5 of 10 percent), and smoothelin (approx. 3 of 10 percent), and miR-214 antagomiR abrogated hypoxia-induced suppression of the contractile phenotype. We also found that hypoxia-induced hPASMC proliferation was significantly attenuated by the anti-miR (approx. 2-fold less compared to hypoxia control). Further, anti-miR-214 also restored PAH-PASMCs to a contractile (approx.5 of 10 percent reversal of MEF2C and leiomodin1 expression), and less proliferative phenotype seen during vascular homeostasis.

Conclusions:
Our findings illustrate a key role for miR-214 in modulation of MEF2C-MYOCD-leiomodin1 signaling and suggest that an antagonist of miR-214 could mitigate SMC phenotype changes and proliferation in vascular hyperproliferative disorders including PAH.
Abstract P132: MEF2C-MYOCD and Leiomodin1 Suppression by miRNA-214 Promotes Smooth Muscle Cell Phenotype Switching in Pulmonary Arterial Hypertension
Authors: Sahoo Sanghamitra , Meijles Daniel N , Al Ghouleh Imad , Tandon Manuj , Cifuentes-Pagano Eugenia , Sembrat John , Rojas Mauricio , Goncharova Elena A , Pagano Patrick J
Published: March 2022

Vascular hyperproliferative disorders are characterized by excessive smooth muscle cell (SMC) proliferation leading to vessel remodeling and occlusion. In pulmonary arterial hypertension (PAH), SMC phenotype switching from a differentiated contractile to a synthetic state contributes to disease progression. While SMC contractile phenotype is reportedly maintained by a MEF2C-Myocardin (MYOCD) transcription factor interplay, its molecular control is poorly understood. MicroRNAs (miRs) have emerged as modulators of many cellular processes and some evidence associates them to MEF2C-MYOCD signaling. It is, therefore, plausible that miRs can regulate the synthetic SMC phenotype. We hypothesized that suppression of SMC contractile phenotype in PAH is mediated by miR-214 via repression of signaling by MEF2C-MYOCD and downstream contractile proteins leiomodin1 and smoothelin. Using qRT-PCR, the levels of miR-214 expression were shown to be upregulated in pulmonary artery SMC (PASMCs) from PAH- vs. control human subjects as well as in commercially obtained human PASMC (hPASMCs) exposed to hypoxia (~1.5 fold, p<0.05). These increases in miR-214 were paralleled by downregulation of MEF2C, MYOCD, and SMC-specific contractile proteins, leiomodin1 and smoothelin. MicroRNA-214 overexpression mimicked the PAH profile, downregulating MEF2C (1.00±0.054 vs. 0.696±0.026, p<0.05) and leiomodin1 (1.00± 0.051 vs. 0.281±0.095, p<0.05) protein levels for control vs miR-214 mimic, respectively. Hypoxia significantly reduced expression of SMC-specific contractile proteins leiomodin1, calponin1 (~50%) and smoothelin (~30%), and miR-214 antagomiR abrogated this response. We also investigated whether miR-214 participates in the induction of hPASMC proliferation, and found that hypoxia-induced hPASMC proliferation was significantly attenuated by the anti-miR (~2-fold). Further, anti-miR-214 restored PAH-PASMCs to a contractile (~50% reversal of MEF2C and leiomodin1 expression) and less proliferative phenotype. Our data illustrate a key role for miR-214 in modulation of MEF2C-MYOCD-leiomodin1 signaling and suggest that an antagonist of miR-214 could mitigate SMC phenotype changes in vascular hyperproliferative disorders including PAH.
Abstract P137: Accelerated Arterial Stiffness During the Menopausal Transition - Results from Study of Women’s Health Across the Nation
Authors: Khan Zubair A , Janssen Imke , Mazzarelli Joanne K , Powell Lynda H , Dumasius Andrius , Everson-Rose Susan A , Barinas-Mitchell Emma , Weinstock Perry J , Hollenberg Steven M
Published: March 2022

Background:
Arterial stiffness is an independent marker of cardiovascular disease (CVD). We hypothesized that arterial stiffness would increase in women undergoing menopausal (MP) transition.

Methods:
349 healthy, peri-MP participants from SWAN-Heart, an ancillary study of SWAN (Study of Women’s Health Across the Nation) were evaluated for subclinical atherosclerosis at baseline and again at 2 years. Subjects using hormone replacement (15) or vasodilators (38) were excluded. Radial tonometry waveforms were digitized and a transfer function was utilized to calculate Aortic Augmentation index, adjusted for heart rate of 75bpm (AIx@75). Pulse wave velocity (PWV) was recorded as carotid-to-femoral arterial pulse propagation time. Carotid intima-media thickness (cIMT) was measured by ultrasound. Women were grouped into pre/early-MP (PMP) and late peri/post-MP (LMP). Kruskal-Wallis test was utilized to compare differences among groups.

Results:
Mean duration of follow up was 2.4 years with a retention rate of 69% (203/296). Age at baseline was (mean±SD) 51±3, BMI 29±6; 46 subjects transitioned from PMP to LMP, 82 remained at PMP and 75 at LMP. Peripheral blood pressure (BP) readings did not differ among groups at baseline or follow up (mean, 120/77 mmHg at both). Augmentation index ([[Unable to Display Character: ∆]]AIx@75) changed from 35±6 to 40±6%, for a difference of 5±8, for women transitioning from PMP to LMP vs 34±8 to 34±6% in women staying in PMP and 36±8 to 38±9% in women staying at LMP, p=.04. The trend for [[Unable to Display Character: ∆]]PWV was similar in PMP to LMP, changing from 6.9 to 8.4 m/s ([[Unable to Display Character: ∆]] = 1.5±2.1) compared to PMP, from 7.4 to 8.3 m/s ([[Unable to Display Character: ∆]] = 0.9±1.9) and LMP, from 8 to 8.4 m/s ([[Unable to Display Character: ∆]] = 0.4±2.5), p=.19. There was no difference in cIMT change between all groups; from 0.67 to 0.7 mm ([[Unable to Display Character: ∆]] = 0.03±0.05) in PMP to LMP, from 0.64 to 0.68 mm ([[Unable to Display Character: ∆]] = 0.04±0.05) in PMP and from 0.64 to 0.68 mm ([[Unable to Display Character: ∆]] = 0.04±0.07) in LMP.

Conclusion:
Augmentation index, a physiologic measure of arterial stiffness, increased through the menopausal transition. There was no significant difference in cIMT change, showing that physiological changes occur before detectable morphologic changes in arterial vasculature in this setting. Changes in arterial stiffness occur first and may mediate the increased CVD risk in women undergoing menopause.
Abstract P139: Ft3 Level Correlates with Arterial Stiffness and Systolic Blood Pressure: A Cross-sectional Study
Authors: Wang Wenyao , Carrillo-Sepulveda Maria A , Yang Yuejin , Tang Yi-Da
Published: March 2022

Background:
Previous studies have suggested that thyroid dysfunction was associated with numerous risk factors of cardiovascular diseases, but whether thyroid function status could influence arterial stiffness and blood pressure (BP) has not been investigated sufficiently.

Methods:
From January to December 2011, we enrolled 1732 patients who took physical examination in our center consecutively. The exclusion criteria were: previous histories of cardiovascular and cerebrovascular diseases, current usage of any anti-hypertensive drugs, anti-thyroid drugs, or thyroxin. All patients underwent testing for thyroid function status, brachial-ankle pulse wave velocity (baPWV), and ankle brachial index (ABI). We investigated the relationship between thyroid hormone levels, arterial stiffness markers (baPWV and ABI) and BP.

Results:
There were 97 patients (5.6%) who had thyroid dysfunction, including low-T3-syndrome (11 patients, 0.64%), subclinical hypothyroidism (51 patients, 2.94%) and clinical hypothyroidism (19 patients, 1.10%), subclinical hyperthyroidism (16 patients, 0.92%). After adjusting for conventional risk factors (age, gender, smoking, diabetes mellitus, dyslipidemia, previous hypertension), free triiodothyronine (FT3) was negatively correlated with baPWV (r=-0.482, P<0.001) and positively correlated with ABI (r=0.290, P<0.001), indicating that the lower FT3 level correlates with arterial stiffening. FT3 also had a moderate negative correlation with systolic BP (r=-0.375, P<0.001). TSH was significantly associated with baPWV (r=0.327, P=0.002), but it did not have any relationship with ABI and BP. No significant correlation was found between other thyroid hormone levels (TT4, FT4 and TT3), baPWV, ABI and BP.

Conclusions:
Lower FT3 and higher TSH were associated with arterial stiffening markers. In addition, FT3 is negatively correlated with systolic BP. These results indicate that thyroid hormone levels play an important role in arterial stiffness and hypertension. Further study is warranted to investigate whether thyroid hormone therapy could benefit people with arterial stiffening and hypertension.