bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract P179: Placental Ischemia-Induced Hypertension is Attenuated in Melanocortin-4 Receptor (MC4R)-Deficient Pregnant Rats
Authors: Spradley Frank T , Granger Joey P
Published: December 2018

The incidence of preeclampsia (PE), a pregnancy-specific disorder of new-onset hypertension that is linked to placental ischemia, has increased over the last several decades. While obesity is a major risk factor for PE, the mechanisms whereby obesity impacts placental ischemia-induced hypertension are not fully understood. We recently reported that placental ischemia-induced hypertension is partially dependent upon an intact sympathetic nervous system (SNS). In various hypertensive models including obesity, blockade of MC4R signaling leads to increased food intake and body weight but protects against the development of hypertension via suppression of the SNS. Although blockade of MC4R may lead to increased body weight during pregnancy, we tested the hypothesis that placental ischemia-induced hypertension is attenuated in obese MC4R-deficient pregnant rats. Wild-type (WT) and MC4R heterozygous-deficient rats were mated with genotype-matched males at ~12 wks old. All rats were maintained on NIH31 chow diet. On gestational day 14, rats were subjected to chronic placental ischemia via the reduced uterine perfusion pressure (RUPP) procedure or Sham surgery. By day 19, conscious mean arterial blood pressure (MAP, carotid catheter) was elevated in RUPP WT (N=21) over Sham WT rats (N=14) (117±2 vs. 101±1 mmHg, P<0.05) with reduced average fetus (1.71±0.04 vs. 1.89±0.05 g, P<0.05) but no change in average placenta weights (0.46±0.02 vs. 0.47±0.01 g), respectively. In Sham MC4R-def vs. Sham WT, respectively, there was increased (P<0.05) body weight (366±8 vs. 337±6 g), total body fat mass (72±4 vs. 44±2 g), and circulating leptin adipokine levels (6.9±0.7 vs. 3.3±0.3 ng/mL) but with similar average fetus (1.92±0.02 vs. 1.89±0.05 g) and placenta weights (0.49±0.01 vs. 0.47±0.01 g). Even though blood pressure was elevated in Sham MC4R-def (109±2 mmHg, P<0.05) over Sham WT, the hypertensive response to RUPP was attenuated in MC4R-def rats (114±2 mmHg). RUPP did not alter leptin levels in MC4R-def rats (6.4±1 ng/mL). In conclusion, these data suggest that MC4R heterozygous-def pregnant rats are obese and have higher blood pressure during pregnancy. However, placental ischemia may require a full complement of MC4R expression to elicit a hypertensive response.
Abstract P180: Cardiac Small Vessel Imaging by Light Sheet Microscopy and Micro CT - Discovering the Missing Link Between Preeclampsia and Higher Risk for Further Cardiovascular Disease
Authors: Kraeker Kristin , Haase Nadine , Herse Florian , Verlohren Stefan , Heuser Arnd , Sporbert Anje , Richter Matthias , O'Driscoll Jamie , Thilaganathan Basky , Mueller Dominik N. , Dechend Ralf
Published: December 2018

Introduction:
In preeclampsia, symptoms like high blood pressure and albuminuria are caused by a state of anti-angiogenic and immune imbalance resulting in endothelial dysfunction. The evaluation of smaller vessels is a challenge, but clinically of increasing importance.

Objective:
We want to examine whether the increased risk for postpartum maternal cardiovascular disease after preeclamptic pregnancy is resultant from microvascular changes in connection with the structural remodelling processes.

Methods:
We compared echo data from a human cohort with data from our transgenic animal model (hAGTxhRen) after preeclamptic pregnancy. In addition, we investigated cardiac changes in gene (qPCR) and protein expression levels (ELISA, IHC staining) in maternal rats, as well as alterations in microvascular 3D remodeling using LSFM and Micro CT.

Results:
We were able to show that the echo changes in our transgenic rat model are comparable to human data. Basic parameters like ejection fraction (human 0,91; animal 0,86), end-systolic volume (human 1,26; animal 1,22) or heart rate (human 1,13; animal 1,16) are altered at the end of pregnancy in same direction. Also markers of hypertrophy like relative wall thickness (human 1,19; animal 1,21) are changed. With the help of speckle trackle analysis, a more sensitive method to detect subclinical changes in hearts functionality, both groups show a reduction of global longitudinal strain (human 0,75; animal 0,6) and strain rate (human 0,81; animal 0,73). The microvasculature and entire vascular network has been visualized so far only in cleared mouse brains and partially in adult mouse hearts. Here, we present the 3D network of lectin-labeled blood vessels in cleared adult rat hearts with the analysis of cardiac small vessels with regard to branching points, vessel length and up to a diameter of 6μm.

Discussion:
Preeclampsia leads to a weakened functionality in postpartum hearts, in a human cohort as well as in our transgenic animal model. Studies are underway to quantify coronary microvascular pathology as a possible missing link between preeclampsia and higher risk for further cardiovascular disease. (values show postpartum change compared to non-pregnant)
Abstract P184: Coordinated Control of Cardiac Contractility By Multiple RGS Proteins
Authors: Bernadyn Tyler F , Marshall Jackson G , Dahlen Shelby A , Meleka Matthew M , Owens Elizabeth A , Osei Owusu Patrick
Published: December 2018

Signaling via heterotrimeric G proteins is critical to maintaining cardiovascular homeostasis. Abnormal G protein signaling due to the loss of G protein regulation by proteins such as regulators of G protein signaling (RGS) proteins is implicated in several cardiovascular disorders such as hypertension, cardiac hypertrophy, and heart failure. RGS act as GTPase activating proteins (GAPs) to control the kinetics and amplitude of G protein signaling. Multiple RGS proteins are prominently expressed in the cardiovascular system; however, it is unknown whether their activities/functions are coordinated to control G protein signaling and cardiovascular function. This study used mice concurrently lacking RGS2 and 5 (
Rgs2/5
dbKO) to determine how the dual loss of potent GAPs for Gq/11 and Gi/o class G proteins affects cardiovascular function. Blood pressure and heart rate in conscious, freely moving mice were monitored via radiotelemetry. Surgical implantation of the radiotelemetry device induced marked systolic blood pressure increase in
Rgs2/5
dbKO mice (WT: 140 ± 6 vs. dbKO: 170 ± 2 mmHg;
p
<0.01) at baseline, which gradually declined but remained elevated above wild type (WT) control level several days later. Whereas all WT mice survived the surgery, ~70-80% of male
Rgs2/5
dbKO mice died 72-96 hr post-surgery. When subjected to cardiac stress test using acute dobutamine infusion and echocardiography, male
Rgs2/5
dbko mice showed hypocontractile response relative to WT mice. Freshly isolated ventricular cardiomyocytes from male
Rgs2/5
dbKO mice showed decreased fractional shortening (WT: 16.1 ± 4.3 vs. dbKO: 7.4 ± 1.1 %;
p
<0.01) but high calcium transients (WT: 117 ± 20 vs. dbKO: 198 ± 50 au;
p
=0.07) at baseline, and application of electrical field stimulation or the non-selective β-adrenergic receptor agonist, isoproterenol (ISO), triggered premature calcium transients, tachyarrhythmia and death of cells from
Rgs2/5
dbKO mice. Interestingly, cells from mice harboring just one copy of
Rgs2
(
Rgs2+/-,Rgs5-/-
) but not
Rgs5
(
Rgs2-/-,Rgs5+/-
) were resistant to low-dose ISO-induced arrhythmia. These results together suggest that RGS2 and 5 coordinate their activity to control cardiomyocyte excitation-contraction coupling and normal cardiac rhythm.
Abstract P186: Reversion of Severe Cardiac and Renal Dysfunction With Surgical Correction of Atypical Aortic Coarctation in Adult Patient: Case Report
Authors: Bortolotto Alexandre L , Filho Rogerio B , Moreira Everton L , Dallan Luis A , Bortolotto Luiz
Published: December 2018

Background:
Heart and renal dysfunction are the most frequent complications of arterial hypertension (AH). Secondary causes of AH can lead to severe organ damage, potentially reversible after treatment.

Objective:
To report a rare cause of AH in adult, atypical aortic coarctation, where surgical correction promoted reversion of organ damage.

Report:
Woman, 55 years-old, history of AH for 10 years, worsening of blood pressure (BP) control for the last 10 months, and progressive dyspnea on exertion, reaching dyspnea at rest. Admission: BP=260x110mmHg and acute pulmonary edema. Improvement of clinical condition with intensive measures, and after, the patient showed differences in BP between left (120x100mmHg) and right upper arm (180x110mmHg), systolic murmur on mitral region, absence of pulse in lower limbs. Exams: Serum creatinine (Cr) = 1,7mg/dL; EKG: left chamber overload; Echocardiogram: table; Magnetic resonance angiography of the aorta: severe stenosis of the aortic arch (13.6mm in its largest diameter). Patient was submitted to a bypass (Dacron #18 tube) from the ascending to the descending segment of the aorta. In the post-operatory period the patient improved renal function (Cr 1.0mg/dl) and controlled BP under use of 1 drug. One month after surgery, patient maintained BP adequately controlled significantly improved echocardiographic parameters (table). In long-term follow-up, patient remained asymptomatic with BP=124x72mmHg, and normal renal and cardiac function, under use of 3 antihypertensive drugs.

Conclusion:
Atypical aortic coarctation at adult age can lead to severe cardiac and renal impairment, which is reversible after surgical correction.
Abstract P188: Adipose-Specific Deletion of Angiopoietin-like 4 Protects Against High Fat Diet Induced Dyslipidemia
Authors: Spitler Kathryn M , Cushing Emily M , Shetty Shwetha , Davies Brandon S
Published: December 2018

Hypertriglyceridemia is a risk factor for the development of hypertension and diabetes. Plasma triglycerides (TGs) are hydrolyzed by lipoprotein lipase (LPL) to release free fatty acids for uptake into tissues. A circulating inhibitor of LPL is angiopoietin-like 4 (A4). A4 is expressed in liver and adipose and is present in circulation. In both humans and mice, A4 deficiency reduces plasma TGs. We previously reported that A4 deficient mice clear more TGs from plasma to adipose tissue. However, it is not clear if this effect is mediated by circulating ANGPTL4 or from local, adipose-derived A4. In this study we sought to characterize the effects of adipose- or liver-specific deletion of A4 on lipid homeostasis. Liver-specific or adipose-specific A4 knockout mice (A4-LivKO/AdipoKO) were generated by crossing A4 floxed mice (A4-fl/fl) with albumin-cre mice or adiponectin-cre mice, respectively. At 8 weeks of age, mice were randomized to either a normal chow diet (NCD) or a high fat diet (60% fat/kCal) for 12 weeks. TG levels (mg/dL) were significantly decreased in A4-AdipoKO mice at baseline (90±4) vs. A4-fl/fl (132±5). No differences were seen in A4-LivKO vs. A4-fl/fl mice (132±3 vs 130±4). After 12 weeks on diets, the A4-AdipoKO mice on NCD (92±6) and HFD (73±6) maintained lower plasma TG levels than A4-fl/fl mice (145±12) on NCD. No differences were seen in TG levels between A4-LivKO mice on HFD or NCD compared to A4-fl/fl mice. TG uptake was increased into adipose tissue of HFD fed A4-AdipoKO [(%-injected dose/mg tissue) (epididymal (eWAT) 0.82± 0.2 and brown adipose tissue (BAT) 1.74± 0.6)] compared to floxed controls on HFD (eWAT 0.44± 0.3 and BAT 0.93± 0.9). Liver-specific loss of ANGPTL4 had no differences in TG uptake from floxed controls on HFD. Overall, we anticipate that identifying tissue-specific contributions will be valuable in informing therapeutic approaches for treating hypertriglyceridemia.
Abstract P189: The Prorenin Receptor and its Soluble From Stimulate Hepatic Triglyceride and Cholesterol Metabolism in Male Mice
Authors: Gatineau Eva , Yiannikouris Frédérique
Published: December 2018

Non-alcoholic fatty liver disease is strongly associated with obesity and metabolic syndrome. Our laboratory previously demonstrated that mice lacking adipose prorenin receptor (PRR) have elevated hepatic triglycerides and cholesterol contents. In addition, lack of adipose PRR induced a counter regulatory increase of hepatic PRR and plasma soluble PRR (sPRR). Therefore, our study aimed to determine whether PRR and sPRR stimulated endogenous hepatic triglycerides and cholesterol synthesis. PRR floxed male mice fed a standard diet were injected with a single dose of an adeno-associated virus thyroxine-binding globulin Cre (KO, n=5) or saline (CTL, n=6). Body weight were measured weekly for 3 weeks. The deletion of PRR in liver did not change body weight curves. Hepatic levels of TG were significantly decreased in liver PRR KO mice compared with control mice (CTL, 615 ± 196 mg/g prot; KO, 114 ± 18 mg/g prot; P=0.047). PRR KO decreased significantly PPARγ gene expression suggesting that PRR positively regulates TG contents through PPARγ pathway. Surprisingly, similar to adipose PRR KO, liver PRR KO increased significantly plasma sPRR (CTL, 5942 ± 199 pg/ml; KO, 8903 ± 559 pg/ml; P=0.001) and hepatic total cholesterol levels (CTL, 40 ± 3 mg/g prot; KO, 102 ± 6 mg/g prot; P<0.0001). This elevation was associated with an increase in SREBP-2 and HMGCoA-reductase gene expression. To determine whether sPRR stimulated cholesterol synthesis, HepG2 cells were treated with sPRR and sPRR was infused in C57BL/6J mice (n=7 to 8 mice/group). Results showed that sPRR significantly up-regulated hepatic SREPB2 mRNA expression in HepG2 cells (Veh, 1.00 ± 0.04; sPRR, 1.36 ± 0.08; P=0.009) and in the liver of mice (Veh, 1.01 ± 0.07; sPRR, 1.23 ± 0.07; P=0.04). In conclusion, our results demonstrated that hepatic PRR positively regulated TG contents through a PPARγ-dependent mechanism and that sPRR stimulated cholesterol synthesis via SREBP-2 pathway. Futures studies will investigate the mechanism by which PRR and sPRR modulate lipid metabolism.
Abstract P192: Trpc6 Deficiency Causes Obesity, Impaired Glucose Tolerance and Leptin Resistance
Authors: Wang Zhen , do Carmo Jussara M , da Silva Alexandre A , Moak Sydney P , Bailey Kandice C , Hall John E
Published: December 2018

Transient receptor potential cation channel subfamily C member 6 (TPRC6) is a receptor-operated cation channel that contributes to changes in cytosolic free Ca
2+
concentration. TRPCs have been suggested to play a role in the regulation of neurite outgrowth, synapse formation, and neuronal survival. However, the role of TRPC6 in controlling metabolic and cardiovascular functions is still unclear. We therefore investigated the impact of TRPC6 on regulation of energy balance, metabolic and cardiovascular functions and anorexic responses to leptin in male and female TRPC6 knockout mice and control B6/129 mice, including assessment of body weight (BW), food intake (FI), body fat/lean composition, responses to acute leptin injection, glucose tolerance tests (GTTs), and blood pressure (BP) and heart rate (HR). TRPC6 null mice of both sexes were heavier than control mice from 6 to 16 weeks of age when fed a standard diet (39.5 ± 1.5 vs 31.1 ± 1.2 g in males and 31.1 ± 1.2 vs 22.9 ± 1.0 in females at 16 weeks of age, n=7,
p
<0.05). EchoMRI scans showed that the higher body weight in TRPC6 null mice was mainly due to increased body fat compared to controls (30.0 % vs 22.6 % of fat mass/BW in males and 32.7 % vs 21.0 % in females at 16 weeks of age), and was associated with increased daily FI (3.4 ± 0.1 vs 2.7 ± 0.2 g in males and 2.7 ± 0.1 vs 2.0 ± 0.1 in females at 16 weeks of age, n=7,
p
<0.05). Acute leptin injections (5 mg/kg, i.p. at 18 weeks of age, n=7) significantly reduced 24-hr FI by 41 % and 32 % in male and female control mice, while only an 8 % and 18 % reduction in FI were observed in male and female TRPC6 null mice. At 20 weeks of age, male and female TRPC6 null mice also exhibited impaired glucose tolerance during GTT compared to controls (AUC: 16,138 ± 1,877 vs 8,016 ± 1,476 mg/dL x 120 min in males, and 6,152 ± 1,173 vs 1,975 ± 512 mg/dL x 120 min in females, n=7,
p
<0.05). BP and HR measured by telemetry for 5 consecutive days at 22 weeks of age were similar between groups. Our results indicate that TRPC6 plays an important role in normal control of food intake, body weight and glucose homeostasis as well as for leptin’s anorexic action. Although TRPC6 deficiency caused obesity and metabolic abnormalities, BP and HR did not increase.
Abstract P194: Body Silhouette Trajectories Across the Lifespan and Vascular Aging: The Paris Prospective Study 3
Authors: van Sloten Thomas , Boutouyrie Pierre , Lisan Quentin , Tafflet Muriel , Thomas Frédérique , Guibout Catherine , Climie Rachel , Pannier Bruno , Sharman James , Laurent Stéphane , Jouven Xavier , Empana Jean-Philippe
Published: December 2018

Background:
Vascular aging, i.e. accumulation of functional and structural changes of vessels throughout life, is a major contributor to cardiovascular disease. It can be quantified by higher carotid stiffness, intima-media thickness and diameter, and hypertension. Weight gain across the lifetime may be an important, modifiable determinant of vascular aging.

Aim:
To assess lifetime body silhouette trajectories (a marker of weight change across the lifespan) in relation to vascular aging in late adulthood.

Methods:
We used cross-sectional data from a community-based study (n=8,083; age 59.4; 38.6% women). A linear mixed model was used to assess trajectories of recalled body silhouettes from age 8 to 45. We assessed carotid stiffness, IMT and diameter (ultrasonography), resting hypertension (blood pressure ≥140/90 mmHg or use of antihypertensives), and exaggerated exercise blood pressure, a marker of masked hypertension (systolic blood pressure ≥150 mmHg during submaximal exercise).

Results:
We identified 5 trajectories: lean-stable (32%), lean-increase (11%), moderate-stable (33%), lean-marked increase (16%) and heavy-stable (8%). Compared to those in the lean-stable trajectory, individuals in the moderate-stable, lean-marked increase and heavy-stable trajectories had more evidence of vascular aging (Figure).

Conclusions:
Vascular aging was most prominent among individuals who were lean in early life but gained weight during young adulthood, and among those who were heavy in early life and maintained weight. This suggests that prevention of weight gain across the life course, especially during young adulthood, is important to promote healthy vascular aging.
Abstract P196: Fenofibrate Improves Vascular Contractility and Endothelial Function in Diabetic Mice
Authors: Xu Nan , Wang Qin , Wang Qi J , Jiang Shan , Zhou Su H , Zhao Liang , Lai En Y
Published: December 2018

Aim:
To exam the hypothesis that fenofibrate improves vascular endothelial dysfunction via balancing endothelium-dependent relaxation and constriction of aorta in diabetic mice.

Methods:
The streptozotocin-induced diabetic mice were treated with fenofibrate (100 mg/Kg/d, 8 weeks). The responses were assessed from serum and aortic parameters, vascular reactivity by wire myograph and aortic protein expression of endothelial nitric oxide synthase (eNOS), peroxisome proliferator-activated receptor α (PPARα), AMP-activated protein kinase-α (AMPKα) and liver kinase B1 (LKB1) and cyclooxygenase-2 (COX-2) in control and diabetic mice given placebo or fenofibrate.

Results:
Comparing with the control mice (12.2±2.0 μmol/L), serum creatinine increased by 114.8% in diabetic mice (26.2±2.0 μmol/L) and by 66.2% in fenofibrate-treated diabetic mice (21.6±2.8 μmol/L, p<0.01). NO level in aorta was 49.5±8.2 (μmol/g) in diabetic mice and 71.7±5.2 (μmol/g) in fenofibrate-treated diabetic mice (p<0.01). Compared with placebo-treated diabetic group, phosphorylated-eNOS protein in aorta was 3.6±1.0 fold higher (p<0.01) and endothelium-dependent relaxation was increased in fenofibrate-treated diabetic mice (p<0.01). The aorta vasodilation by fenofibrate treatment was reversed with PPARα inhibitor or AMPKα inhibitor. Fenofibrate treatment elevated PPARα expression, subsequently induced LKB1 translocation from nucleus to cytoplasm to activate AMPKα, thus activated eNOS in diabetic aorta (p<0.01). Fenofibrate treatment reduced vascular contractility and expression of COX-2 in aorta from diabetic mice(p<0.01). Prostaglandin E2 and thromboxane A2 were 174.9±16.4 and 128.9±4.4 (pg/ml) in diabetic mice, and reduced to 142.7±14.1 and 105.9±5.7 (pg/ml) in fenofibrate-treated mice (p<0.01).

Conclusion:
Fenofibrate treatment in diabetic mice normalizes endothelial function by balancing vascular reactivity via increasing NO production and suppressing vasoconstrictor prostaglandin. Our results suggest that implication of fenofibrate as a putative mediator of diabetic vascular complications.
Abstract P201: Exposure to Stress and Low Salt Diet Increases Blood Pressure in Mice Lacking the Brain-Specific Renin Isoform Renin-b
Authors: Nakagawa Pablo , Gomez Javier A , De Azevedo Cruz Seara Fernando , Sigmund Curt D
Published: December 2018

Previous experience to various hypertensive conditions including psychosocial stress and high or low salt diet enhances the response to subsequent hypertensive challenges. Importantly, the activation of the brain renin angiotensin system (RAS) is required for the sensitization to these hypertensive challenges. We previously reported that the brain-specific renin isoform (Ren-b) tonically inhibits the activation of the brain RAS and the ablation of Ren-b results in brain RAS disinhibition leading to hypertension. Interestingly, we observed a high degree of variability in the blood pressure (BP) between Ren-b KO cohorts that can be attributed to different levels of environmental stress. Thus, BP and heart rate (HR) from five separate experimental cohorts which were subjected to different levels of stress were re-analyzed and compared. In cohorts that were not subjected to any surgical procedures, Ren-b KO mice housed in a stressful environment exhibited higher systolic BP (cohort 1, KO: 130 ± 1 vs WT: 122 ± 1 mmHg, p<0.05) in comparison with animals housed in a quiet and more regulated environment (cohort 2; KO: 116 ± 1 mmHg vs WT: 116 ± 1). BP was selectively higher in Ren-b KO mice subjected to a surgical procedure required to study a BP mechanism (Cohort 3, KO: 137 ± 2 vs WT: 117 ± 3, p<0.05; Cohort 4, KO: 139 ± 2 vs WT: 112 ± 6, p<0.05; Cohort 5, KO: 137 ± 3 vs WT: 113 ± 3 mmHg, p<0.05). Similarly, elevation of BP was associated with increased HR. Altogether this reanalysis of pre-existing data suggests that Ren-b KO mice might be sensitive to environmental or surgical stressors. Next, we hypothesized that Ren-b KO mice are sensitized to high salt diet, thus BP was acquired in WT or Ren-b KO fed low or high salt diet (0.3% and 4.0% NaCl, respectively) for a week in a quiet environment. On high salt diet, Ren-b KO did not manifest an increase in mean BP (WT: 113±2 vs KO: 109±2 mmHg, p=0.18), indicating that Ren-b KO are not salt-sensitive. In contrast, on low salt diet Ren-b KO exhibited modest increase in mean BP compared to WT (WT: 108±2 vs KO: 113±1 mmHg, p<0.05) indicating that Ren-b KO might be sensitive to conditions that exacerbate activation of peripheral RAS. We conclude that disinhibition of brain RAS by the ablation of Ren-b leads to exaggerated response to hypertensive challenges.
Abstract P214: Serum Glucocorticoid Kinase 1 and Sympathetic Nerve Activity in Salt-Sensitive Hypertension.
Authors: Van Beusecum Justin P , Barbaro Natalia R , Xiao Liang , Pandey Arvind K , Harrison David G , Kirabo Annet
Published: December 2018

Salt-sensitive hypertension affects nearly 50% of the population and reducing salt intake decreases blood pressure and cardiovascular events in the general population. The precise mechanism of how dietary salt contributes to blood pressure (BP) elevation, renal injury, and cardiovascular disease remains unclear. Substantial evidence supports a role of increased sympathetic output in salt-dependent hypertension and this promotes renal inflammation and dysfunction. It has been shown that salt accumulates in the interstitium of hypertensive humans and animals and drives immune cells toward a proinflammatory phenotype through the salt sensing kinase serum/glucocorticoid kinase 1 (SGK1). We have also shown an important role of monocytes and monocyte-derived dendritic cells in hypertension. In this study, we tested the hypothesis that SGK1 in myeloid CD11c
+
cells promote salt-sensitive hypertension by increasing sympathetic outflow. To test this hypothesis, we created mice lacking SGK1 in CD11c
+
cells (SGK1
CreCD11c
mice) and used SGK1
fl/fl
mice as controls. To induce salt-sensitivity, mice received 0.5 mg/ml of N-Nitro-L-arginine methyl ester hydrochloride (L-NAME) in the drinking water for 2 weeks. This was followed by a 2-week washout period and then a 4% high salt diet for 3 weeks. BP was monitored using telemetry. We found that BP elevation during high salt feeding was significantly attenuated in SGK1
CreCD11c
mice compared to SGK1
fl/fl
mice (125 ± 1 vs. 141 ± 1 mmHg; p < 0.01). SGK1
CreCD11c
mice had a significant reduction in CD45
+
, CD3
+
, CD4
+
CD8
+
and CD19
+
cells in the kidney compared to SGK1
fl/fl
mice as assessed by flow cytometry (p < 0.05). Interestingly, on high salt, SGK1
CreCD11c
mice had a significant reduction in heart rate (HR) compared to SGK1
fl/fl
mice (528 ± 2 vs. 592 ± 7 bpm; p = 0.006). Microglial cells of the brain express CD11c, and we found that the reduction in HR was associated with a marked reduction in HR variability in SGK1
CreCD11c
mice compared to SGK1
fl/fl
mice (1.65 ± 0.38 vs. 3.00 ± 0.57; p = 0.0018), indicating a reduction in sympathetic outflow. Our data indicate that SGK1 in CD11c
+
myeloid cells and likely those of the central nervous system, modulate BP and sympathetic outflow promoting the pathogenesis of salt-sensitive hypertension.
Abstract P215: Role of Nox4 - H
2
O
2
Production in Cctivation of mTORC1 in Salt-Induced Hypertension and Kidney Injury in Dahl S Rats
Authors: Kumar Vikash , Kurth Theresa , Cowley Allen
Published: December 2018

Nox4 is the most abundant NADPH-oxidase isoform in the kidney. It predominantly releases H
2
O
2
and has been shown to contribute importantly to hypertension and renal injury in Dahl salt-sensitive (SS) rats fed a high salt diet. Recently, we reported that systemic inhibition of mTORC1 with chronic rapamycin treatment reduces salt-induced hypertension and kidney injury in SS rats. In the present study, we hypothesized that elevations of H
2
O
2
would activate mTORC1 and that reduction of Nox4 related H
2
O
2
production would reduce the mTORC1 contribution to salt-induced hypertension in SS rats. We first examined the effect of H
2
O
2
on mTORC1 activity
in vivo
by comparing the activity of mTORC1 in SS rats which have high levels of H
2
O
2
to SS-Nox4 knockout (SS
Nox4–/–
) rats which have low levels of renal H
2
O
2
. As analyzed by Western blot, pS6
S235/236
/S6 (index of mTORC1 activity) was found to be significantly lower (P<0.05) in the renal cortex of SS
Nox4-/-
rats indicating that Nox4 generated H
2
O
2
was upstream in the mTORC1 pathway. We then examined whether systemic inhibition of mTORC1 with chronic rapamycin treatment would fail to reduce salt-sensitivity and kidney injury in nine week old male SS
Nox4-/-
rats. It was found that mean arterial pressure (MAP) of SS
Nox4-/-
rats treated with rapamycin (I.P.,1.5 mg/kg/day) fed a high salt diet (4.0%NaCl) for 21 days was not significantly different to vehicle treated SS
Nox4-/-
rats on the final day 21 (150 ± 7; n=8; vs 145 ± 6 mmHg; n=5; P>0.05). Neither did rapamycin treatment have a significant effect on kidney injury in SS
Nox4-/-
rats compared to vehicle treated rats determined at day 21 as reflected by albumin excretion rate (8 ± 2 vs 20 ± 13 mg/day, respectively; P>0.05). Rapamycin treated SS
Nox4-/-
rats exhibited a slow rate of body weight gain (302 ± 14 g at control and 313 ± 12 g at 21 days; P>0.05) compared to vehicle treated rats (316 ± 9 g at control and 366 ± 15 g at 21 days; P<0.05), although average food intake did not differ significantly (P>0.05) between the two groups. We conclude that Nox4 - H
2
O
2
production contributes importantly to mTORC1 activation leading to blood pressure salt-sensitivity, and renal injury in SS rats.
Abstract P222: Oxidative Stress Causes Salt Sensitive Hypertension via Activation of Na
+
-Cl
–
Cotransporter (ncc)
Authors: Wang Conghui , Shimosawa Tatsuo , Latapati Reheman , Koid SuangSuang , Alimila Yeerbolati , Liu BeiBei , Ogura Sayoko , Mori Fumiko , Fujita Toshiro
Published: December 2018

Background:
We have reported that blockade of nitric oxide synthesis by L-NAME causes salt-sensitive hypertension via activating NCC. which was confirmed by the finding that systemic administration of L-NAME failed to cause salt-sensitive hypertension in NCC deficient mice. In vitro study showed when mDCT cells are treated with L-NAME, NCC phosphorylation is increased and NO donner reduced its phosphorylation. However, the precise direct effect of NO on NCC phosphorylation is not clear. In this study we clarified the role of oxidative stress but not angiotensin signaling in NCC phosphorylation

Method:
The passages 3 - 15 of mDCT cells were used in this study. The cells were starved 1 hour before the experiment. L-NAME (10-40 μM), SOD mimetic 4-Hydroxy-TEMPO (10 μM), NO donor sodium nitroprusside, (SNP 100 μM - 2 mM), pSPAK inhibitor STOCK2S-26016, and Losartan (10 μM) were used to treat the mDCT cells for 10 min and 30 min. Oxidative stress was measured by ROS production using the lucigenin method. To confirm the oxidative stress function, we performed vivo experiment with SOD mimetics--4-hydroxy-TEMPO. The C57BL/6J Kwl mice received implantation of subcutaneous osmotic mini-pump containing TEMPO (20 mg/kg/day) with 8 % high salt diet and L-NAME (3.7 μM/day in drinking water) for 4 weeks.

Result: L-NA
ME increased oxidative stress by ROS production (2.0 ± 0.15 times higher than control) and p-SPAK signaling, which was normalized by TEMPO, an SOD mimetic and p-SPAK inhibitor. In contrast, AT1R inhibitor Losartan did not affect L-NAME-induced activation of NCC in mDCT cells. Moreover, the function of TEMPO was also confirmed in vivo study, which attenuated the L-NAME- induced increases in superoxide, mean BP (HS 108.7 ± 3.7 mmHg; HSL 130.5 ± 2.9 mmHg; HSL + TEMPO 116.0 ± 1.7 mmHg) and p-NCC expression in the C57BL/6J mice.

Conclusion:
Oxidative stress - pSPAK pathway plays a pivotal role in the NO-induced inhibition of NCC.
Abstract P229: Effect of Management of Hypertensive Susceptible Population: A Case-Control Study in Shanghai, China
Authors: Tongtong Liang , Dongli Xu , Jinling Zhang
Published: December 2018

Objective:
The management of hypertensive susceptible population was carried out by community family physician in Minhang district since 2011. The management includes health education, blood pressure measurement, and so on.This study was to explore the effect of the management of hypertensive susceptible population under community family physician model.

Methods:
A computerized database of hypertensive susceptible population enrolled in management of community health service centers in Minhang District,Shanghai from 2011 to 2018 was used. According to whether the management status was transferred to patients with hypertension, the subjects were divided into case group and control group. We compared the time of management, demographic characteristics, lifestyle, family history, and disease history between two groups. Univariate analysis was performed by t test and χ
2
test, and Logistic regression analysis was used for multivariate analysis.

Results:
A total of 18163 subjects were enrolled, and 3623 cases were transferred to hypertension patients (case group), accounting for 19.9%(3623 of 18163). The average time of management of case group was (25.00±16.84) months, which was lower than that of control group (38.56±18.45) months (P < 0.001). The results of Logistic regression analysis showed that time of management (OR=0.953,95%CI:0.951-0.955) and exercise (OR=0.655,95%CI:0.554-0.775) were the protective factors for hypertension; the history of diabetes (OR=2.920,95%CI:2.472-3.450), the history of coronary heart disease (OR=1.844,95%CI:1.436-2.367), drinking (OR=1.410,95%CI:1.131-1.757), smoking (OR=1.356,95%CI:1.172-1.570), family history of hypertension (OR=1.226,95%CI:1.121-1.342), age (OR=1.043,95%CI:1.040-1.047), BMI (OR=1.023,95%CI:1.014-1.032) and pulse pressure difference (OR=1.015,95%CI:1.010-1.021) were risk factors for hypertension.

Conclusions:
The management of hypertensive susceptible population under community family physician model can reduce the risk of hypertension and is important for protecting hypertensive susceptible population, after removing the influence of age, BMI, disease history, family history and lifestyle.
Abstract P233: Blood Pressure Variability and Rhythmicity in Adolescents With Spurious Hypertension
Authors: Obrycki Lukasz , Feber Janusz , Niemirska Anna , Litwin Mieczyslaw
Published: December 2018

Objective:
Spurious hypertension (sHTN) is characterized by elevated systolic blood pressure (BP) in peripheral arteries but normal central BP. The BP variability and characteristics of BP rhythms in sHTN is unknown. We therefore analyzed BP variability and rhythmicity in adolescents with sHTN in comparison with other forms of hypertension (HTN).

Patients and Methods:
We studied 265 children (54 girls; 15.1 ±2.5 years) referred with arterial HTN, in whom secondary HTN was excluded. We assessed BP levels - office BP, ABPM, central systolic BP (cSBP), systolic BP average real variability (SBPARV) and mean MAP rhythmicity (amplitudes and acrophases). Anova with TukeyHSD post hoc test was used to compare groups. One tailed t-test was used to compare MAP amplitudes and acrophases with normative values for children with Bonferroni correction for multiple tests.

Results:
115 subjects had white coat hypertension (WCH; office BP>95
th
perc+ABPM<95
th
perc), 25 had ambulatory prehypertension (preHTN; office BP<95
th
perc+ABPM<95
th
perc+ABPM load >25%), 43 had sHTN (office BP+ABPM >95th perc+cSBP<95
th
perc), 82 had true hypertension (tHTN; office BP+ABPM+cSBP >95th perc. There were no significant differences between groups in age, weight and BMI. The median SBPARV was 8.87 in WCH, 8.63 in preHTN, 9.25 in sHTN and 9.28 in tHTN (not significantly different between groups). The prevalence of 24h rhythms ranged from 77 to 88%, not significantly from norms or among groups. The prevalence of 12h rhythms was not significantly different compared to norms and ranged from 50 to 60% (NS among groups). There were also no significant differences in the prevalence of 8h and 6h rhythms. All groups had significantly decreased 24h amplitudes compared to norms, but not significantly different among groups. There were no differences in 12h, 8h and 6 amplitudes between WCH, preHTN, sHTH, tHTN and norms. 24h acrophases were significantly prolonged in sHTN, 12h acrophases were significantly prolonged in preHTN, sHTN and tHTN.

Conclusions:
Adolescents with spurious hypertension have significantly lower prevalence of 24h rhythms, decreased 24h amplitudes and prolonged 24h and 12h acrophases suggesting increased sympathetic drive similarly to patients with other hypertensive phenotypes.
Abstract P236: Sodium Accumulates in the Skin of Patients and Mice With Psoriasis
Authors: Markó Lajos , Wild Johannes , Rakova Natalia , Balogh András , Opitz Elisa , Linz Peter , Birukov Anna , Wilck Nicola , Dechend Ralf , Titze Jens , Kleinewietfeld Markus , Krause Andreas , Kokolakis Georgios , Philipp Sandra , Luft Friedrich C , Boschmann Michael , Kelm Marcus , Kühne Titus , Karbach Susanne , Müller Dominik N
Published: December 2018

Sodium can be buffered in the skin, which mechanism is altered during aging and in certain diseases such as hypertension. High salt environment can promote autoimmunity by expanding pathogenic IL-17 producing T helper (Th17) cells. Psoriasis is a relapsing and remitting inflammatory autoimmune disease affecting the skin and joints and involves proinflammatory Th17 cells. Here we tested the hypothesis if psoriatic skin has a higher sodium content in mice and humans. We used two psoriasis mouse models; the K14-IL-17A
ind/+
mice overexpressing IL-17A in K14-positive keratinocytes and the imiquimod (IMQ) mouse model by applying 62.5 mg IMQ cream (5%) on the shaved back and ears of FVB/N mice for 5 days daily. End of the study skins of mice were collected, weighted, dried and ashed to measure water and sodium content. Additionally, skin sodium and water content were measured in psoriasis patients and aged matched healthy controls by non-invasive
23
Na-MRI on non-affected flexor site of the lower leg and by
23
Na-spectroscopy to compare affected and non-affected sites of the leg. K14-IL-17A
ind/+
mice had significantly higher sodium content compared to control IL-17A
ind/+
mice (0.191±0.021 vs. 0.137±0.023 mg/g dry weight) together with an elevated water content. IMQ-treated back skin had significantly higher sodium content compared to untreated ventral skin of the same mice (0.175±0.023 vs. 0.143±0.014 mg/g dry weight), whereas sham mice had a significantly lower content in both regions (0.116±0.010 vs. 0.107±0.005 mg/g dry weight). IMQ treatment led to significant expansion of IL-17 producing γδT cells in the skin, regional lymph nodes and in the spleen with typical skin lesions. Patients with psoriasis area and severity index (PASI) >5 had significantly higher sodium content in the skin compared to those with lower PASI or with healthy controls (17.73±1.52 vs. 14.32±1.54 vs. 14.30±2.59 AU, respectively); this elevation was water coupled. PASI significantly correlated with skin sodium content (Pearson’s r=0.598, P<0.001). Additionally, patients with PASI>5 has higher sodium content in the affected skin compared to non-affected skin of the same patient. Data from animal models and humans argue for higher sodium accumulation in the inflamed skin.
Abstract P237: Elucidation of Blood Pressure Control Function of Essential Hypertension Candidate Gene Lpin1
Authors: sumida koichiro , Hirawa Nobuhito , Yatsu Keisuke , Ehara Yousuke , Oki Yuki , Fujita Megumi , Fujiwara Akira , Kobayashi Yusuke , Wakui Hiromichi , Umemura Satoshi , Tamura Kouichi
Published: December 2018

We have found that the LPIN1 is the candidate gene for essential hypertension (EHT) by genome-wide association analysis in 2007. Mice characterized by loss of body fat, fatty liver, insulin resistance and peripheral neuropathy were discovered in nature and named fatty liver dystrophy (fld) mice in 2000. Recently, the fld mice were shown that they exhibit few LPIN1 genes, and the LPIN1 gene was proved as the cause of phenotype of fld mice. However, the blood pressure (BP) and the alteratios of the vasoactive factors in fld mice have not yet been elucidated. Therefore, to clarify the BP and the effects of LPIN1 gene defeciency, we analyzed the BP and regulatory factors of the fld mice. Furthermore, we transplanted the fat to fld mice, and clarify the importance of adipocytokine on BP in the model mice. The BP were measured by the tail cuff method and the telemetry method. We performe fat grafting or sham surgery on fld mice, and systolic BP (SBP) was measured by tail cuff method 8 weeks after surgery. We confirmed that the SBP of fld mice were higher than that of CO by the tail cuff method( fld 134±5 vs CO 111±4 mmHg P<0.05). Furthermore, SBP of the fld mice were higher than that of CO (fld 167.7±9.8 vs CO 133.5±10.9 mmHg P<0.01) by the telemetry method. It was confirmed that the adrenalin and noradrenalin excretions in the urine of fld mice were higher than those of CO(fld 0.133±0.067 vs CO 0.038±0.013 μg/day P<0.01 )(fld 0.980 ±0.313 vs CO 0.364±0.080 μg/day P<0.001). There is no significant difference in plasma aldosterone concentration between two groups( fld 434±276.7 vs CO 781±469.5 pg/mL n.s).The SBP tended to decrease with the fat trasplantation in fld mice ( fat transplanted 103.7±4.41 vs sham 114.7±7.67 mmHg P=0.056). We confirmed that fld mouse exhibits higher SBP by both tailcuff method and telemetry method. Because the fld mice exhibit higher PR and high levels of CA concentration and excretion in urine, it was suggested that the sympathetic nervous systems of fld mice were activated. Sympathetic nervous system activity may be related to an increase in BP of fld mouse, and further research is under way. Since it showed that BP decreased by fat grafting, the visceral fat are essential for fld mice to BP control. The implications of adipokine and other fats are under consideration.
Abstract P239: Toll-Like Receptors 9 (TLR9) Play a Key Role in Cardiac and Vascular Dysfunction Associated With 2-Kydney 1-Clip (2K1C) Hypertension in Mice
Authors: Rodrigues Fernanda Luciano , Omoto Ana Carolina Mieko , Menezes da Costa Rafael , Aguiar da Silva Carlos Alberto , Bomfim Gisele Facholi , Carneiro Fernando Silva , Tostes Rita C. , Fazan Junior Rubens , Salgado Helio C
Published: December 2018

Introduction:
Renovascular hypertension is the leading cause of secondary hypertension and is also involved in the physiopathogenesis of resistant hypertension. The activation of toll-like receptors 9 (TLR9) was demonstrated to contribute to the increase in arterial pressure (AP) in spontaneously hypertensive rats. We hypothesize that the TLR9 are also activated in two-kidney one- clip (2K1C) hypertensive mice, eliciting cardiac and vascular dysfunction; therefore, contributing to the increase in AP.

Methods and Results:
C57BL (WT) and TLR9 knockout (TLR9_KO) mice were anesthetized with isoflurane and submitted to 2K1C hypertension by placing a silver clip (0.12 mm) around the left renal artery. After 4 weeks, the cardiac function of mice was evaluated by echocardiography in anesthetized subjects. Following, the direct AP measurement was carried out and the third-order mesenteric resistance arteries (MRA) were removed for vascular function evaluation. All data were compared to Sham-operated WT mice. The AP was remarkably elevated in 2K1C WT mice (133±2 vs 93 ± 4 mmHg), whereas this increase was partially prevented by the absence of the TLR9 (114±5 mmHg). 2K1C hypertension caused cardiac dysfunction in WT mice, displaying decreased ejection fraction (42.6±3.1 vs 52.7±1.5%), fractional shortening (10.1±0.7 vs 13.7±0.9%), stroke volume (25.9±2.2 vs 37.9±1.9 μL) and cardiac output (9.2±0.8 vs 14.6±0.8 mL/min); however, the 2K1K hypertension did not affect the cardiac function in TLR9_KO mice: ejection fraction (54.5±1.6%), fractional shortening (14.5±1.1%), stroke volume (43.4±4.6 μL) and cardiac output (15.2±1.6 mL/min). Vascular dysfunction, characterized by increased contractile response to phenylephrine (Emax: 167.2±2.2 vs 125.9±3.5%) and reduced relaxation to acetylcholine (Emax: 69.3±1.6 vs 87.6±1.4%) and sodium nitroprusside (pD2: 7.3±0.07 vs 7.9±0.05), was observed in 2K1C WT mice, but not in 2K1C TLR9_KO mice (Emax: 127.7±2.9%, 88.5±2.0% and pD2: 7.78±0.06, to phenylephrine, acetylcholine and sodium nitroprusside).

Conclusions:
TLR9 is involved in both cardiac and vascular dysfunctions of 2K1C mice and also in the increase of AP of this experimental model.
Abstract P243: Vascular Drp1 Mediates Angiotensin II-Induced Cardiovascular Hypertrophy via Global as Well as Selective de novo Protein Synthesis.
Authors: Kawai Tatsuo , Cooper Hannah A , Forrester Steven J , Boyer Michael J , Elliott Kathy J , Preston Kyle , Kimura Yayoi , Sesaki Hiromi , Scalia Rosario , Rizzo Victor , Eguchi Satoru
Published: December 2018

Mitochondrial fission has been implicated in various cardiovascular and metabolic diseases but the role of the mitochondrial fission inducer Drp1 in regulating hypertension and cardiovascular remodeling remains unclear. To study the involvement of Drp1 in cardiovascular hypertrophy, rat aortic vascular smooth muscle cells (VSMCs) are infected with dominant negative (dn) Drp1 adenovirus and stimulated with 100 nM angiotensin II (AngII). In vivo, tamoxiphen-inducible SMMHC-Cre+/- Drp1 floxed mice and control mice are infused with AngII (1000ng/kg/min) for 2 weeks. In VSMCs, AngII-induced mitochondrial fission was attenuated with AT1 blocker RNH6270 (ARB) as well as with dnDrp1 (100 moi). ARB and dnDrp1 treatment also inhibited AngII-enhanced global as well as specific de novo protein synthesis evaluated by SunSet assay. To identify potential mediators downstream of Drp1-mitochondrial fission contributing to VSMC hypertrophy, shotgun proteomic analysis was performed on proteins extracted from AngII or vehicle treated VSMC with dnDrp1 or control adenovirus infection. 22 exclusively regulated proteins induced by AngII and attenuated by dnDrp1 were identified including previously known AngII targets, HMGB1 and PAIRB. In addition, ARB and dnDrp1 inhibited AngII-induced enhancement of mitochondrial ox-phosphorylation and oxidative stress (assessed with mito-timer adenovirus). In vivo, VSMC specific DRP1 silencing suppressed AngII-induced cardiac hypertrophy assessed with heart weight/body weight ratio Mean±SEM: 6.33±0.14 vs 5.25±0.07 (p<0.001) as well as with echocardiogram: LVPWd 1.46±0.02 vs 0.81±0.03 (p<0.001). However, both control and the Drp1 silenced mice developed hypertension when infused with AngII as assessed by DSI radio-telemetry system. In conclusion, these data suggest that Drp1 mediates AngII-induced global as well as specific de novo protein synthesis thereby contributing to cardiovascular hypertrophic remodeling.
Abstract P244: Hypoxic Stimulation of Vasoreparative Functions in Human CD34
+
cells is Mediated by Angiotensin Converting Enzyme-2 and Mas Receptor
Authors: Joshi Shrinidh , Leclerc Estelle , Jarajapu Yagna P
Published: December 2018

CD34
+
stem/progenitor cells have the propensity of re-endothelialization and vascular regeneration. Angiotensin Converting Enzyme-2 (ACE2) generates Angiotensin (Ang)-(1-7), which produces vasoprotection by acting on Mas receptor (MasR). Hypoxic preconditioning has been shown to stimulate vascular repair-relevant functions of CD34
+
cells, which was impaired in MasR-deficient HSPCs. The current study tested the hypothesis that hypoxic stimulation of CD34
+
cell functions are mediated by ACE2 and MasR. CD34
+
cells were isolated from mononuclear cells (MNCs) derived from healthy volunteers (n=46). Cells were exposed to normoxia (20% O
2
) or hypoxia (1% O
2
). Protein and mRNA expressions of ACE2 and MasR were determined. ACE2 activity was determined by using an enzyme-specific fluorogenic substrate, and a selective inhibitor, MLN4760. CD34
+
cells were transduced with lentiviral particles carrying ACE2- or MasR- or scramble-3’-UTR (Scr) fused downstream to firefly luciferase reporter (flr) gene. Co-transfection luciferase assay was performed using MicroRNA, miR-421 or miR-143 with ACE-2 or MasR luciferase construct, respectively. Luciferase activity was determined by using luciferase assay kit. Hypoxia stimulated mRNA and protein expressions of ACE2 and MasR in CD34
+
cells (54.9±4.8% and 51.6±14.7%, respectively, higher than normoxia, n=5, P<0.05) but not in MNCs. No changes in ACE or AT1 receptor expressions were observed. Effects of hypoxia were blocked by 2-methoxyestradiol (0.5μM), an inhibitor of hypoxia-inducible factor-1α (n=5, P<0.01). Luciferase activity was increased by hypoxia in cells expressing ACE2- or MasR-FLR (258±23% and 214±19%, respectively, of Scr, n=6, P<0.001). Effect of hypoxia is recapitulated by vascular endothelial growth factor (100 nM) (n=5, P<0.01 vs untreated) or stromal-derived factor-1α (100 nM) under normoxia (n=5, P<0.05 vs untreated). Axitinib (30 nM), a nonspecific inhibitor of VEGFR, or AMD3100 (10 μM), a specific CXCR4 inhibitor, decreased the luciferase activity in ACE2- or MasR-FLR expressing cells (n=4, P<0.01). Collectively, these observations provide compelling evidence for the hypoxic upregulation of ACE2 and MasR in CD34
+
cells, which contribute to their vasoreparative functions.