bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 085: The Role of CD70 in Vascular Function
Authors: Itani Hana A , Pandey Arvind , Brown Jonathan D , Harrison David G
Published: June 2021

We have recently shown that CD70 is important in formation of effector memory T cells and that these cells seem to enhance the hypertensive response and end organ damage caused by repeated hypertensive stimuli. CD70 has traditionally been identified on antigen presenting cells and is thought to interact with CD27 on T cells, promoting proliferation and memory cell formation. The precise cell types that express CD70 and the role of CD70 on non-immune cells has not previously been investigated. In our prior study, we showed that hypertension is associated with a marked increase in CD70 surface expression on dendritic cells (DCs) and macrophages. To examine the specific effect of DC CD70, we performed adoptive transfer of DCs from hypertensive WT mice to normotensive CD70
-/-
mice and from hypertensive CD70
-/-
mice to normotensive WT mice. The recipient mice were treated with a normally subpressor dose of ang II (140 ng/kg/min). Interestingly the CD70
-/-
recipients were protected from the development of hypertension, despite receiving WT DCs from hypertensive donors. In contrast, the WT recipeints that received CD70
-/-
DCs developed modest hypertension, indicating that non-DC sources of CD70 likely contribute to the hypertensive phenotype. Studies of mesenteric vascular reactivity showed that CD70
-/-
mice have markedly impaired endothelium-dependent vasodilatation to acetylcholine compared to WT mice (43±10 vs 25 ± 3 %) at baseline. In contrast, there were no differences in relaxation responses to sodium nitroprusside. In additional experiments, we showed that human umbillical vein endothelial cells express CD70 mRNA and that this is increased by > 30 fold by laminar shear stress (15 dynes/cm2) compared to oscillatory shear. Finally, using immunohistochemistry, we identified CD70 protein localized to resistance vessels of the kidney of ang II-treated mice. These data identify a new role of CD70 in modulating vascular function independent of its role on antigen presenting cells in memory T cell formation.
Abstract 089: Dopamine D
2
Receptor is Associated with Inverse Salt Sensitivity
Authors: Xu Peng , Gildea John J , Jose Pedro A , Carey Robert M , Felder Robin A
Published: June 2021

Our previous studies of salt sensitivity of blood pressure have demonstrated that approximately 11% of study participants have a paradoxical increase in blood pressure (> or = to 7-mm Hg) on a low NaCl diet (defined as inverse salt sensitivity (ISS)). However the mechanisms responsible for this effect are not known. We demonstrated that single nucleotide polymorphisms (SNPs) in the dopamine type 2 receptor (D
2
R) (RS6276 and 6267) are highly associated with ISS (
P
values of 1.0х10
–2
and 3.8х10
–2
with odds ratios of 0.32 and 0.48 in unadjusted regression models, respectively). The C allele at both sites confers protection. The D
2
R is strongly expressed throughout the cytoplasm of proximal tubule cells in human kidney tissue slices. We also cultured RPTC from the urine from 4 salt resistant (SR) and 3 ISS participants enrolled in our clinical salt sensitivity studies. We hypothesize that D
2
R containing SNPs have altered receptor expression, and altered signaling compared to wild type controls. ISS participants were homozygous variant for the two D
2
R alleles and showed more D
2
R expression than SR RPTC heterozygous variant (HV) for the two alleles (ISS: 1.166±0.059 n=3 vs SR: 0.969±0.024 n=4, P<0.05, t-test). D
2
R expression was increased when the ISS cells were stimulated by a non-selective D
2
R agonist bromocriptine to a greater extent in the D
2
R SNP cell lines (ISS: VEH 1.166±0.059, vs bromocriptine 1.474 ± 0.040, n=3, P<0.05, t-test). Using the ROS reagent assay, dihydroethidium, there was found to be more ROS products in ISS cells than SR cells when stimulated under low salt (ISS: 1.145 ± 0.053, n=3 vs SR: 0.722 ± 0.101, n=4, P<0.05, t-test). We used a highly selective D
2
R agonist (sumanirole) to stimulate wild-type and SNPed cells, and the results demonstrated no effect in the cells with wild type D
2
R but an increase in ROS in cells heterozygous for the D
2
R SNPs (SNP: VEH 38,364±1,266, sumanirole 50,926 ± 3,310, VS WT: VEH 34,562±1,831 sumanirole 34,435 ± 1,614 RFU n=12, P<0.05, t-test) consistent with the higher expression of D
2
R found in ISS urine cells. We hypothesize that SNPs in the D
2
R lead to increased reactive oxygen species which has previously been associated with renal fibrosis and hypertension.
Abstract 094: Pappa2 is Important for Determining the Nephron Number
Authors: Kumar Vikash , Yang Chun , Geurts Aron M , Liang Mingyu , Cowley Allen W
Published: June 2021

Pappa2 is a metalloproteinase which specifically cleaves IGFBP-3 and IGFBP-5 and in turn releases IGF-1. Recently, we have shown that a subcongenic Dahl salt-sensitive (SS) rat strain containing a 0.71 Mbp of chromosome 13 which includes
Pappa2
gene from salt-insensitive Brown Norway (26-P strain) is protected significantly (24 mmHg) from salt-induced hypertension (Cowley et al., 2016). Although it is recognized that Pappa2 modulates development of bone size, cranial cartilage and angiogenesis, its role in kidney development and function is unknown. The present study determined the contribution of Pappa2 to nephron development by comparing SS and 26-P rat strains. It was found that
Pappa2
mRNA expression was 5-fold higher in embryonic kidney (day 20.5) of the salt-resistant 26-P rats compared with age-matched SS rats.
Pappa2
mRNA expression significantly increased with age of kidney reaching a maximum at postnatal day 5 in both strains.
Pappa2
mRNA expression at postnatal day 15 was found to be 9-fold higher in the kidney of 26-P compared with SS strain. Immunohistochemistry studies revealed that Pappa2 co-localized with IGFBP-5 in the ureteric bud indicating that Pappa2 could be important for ureteric branching and nephron endowment. Glomerulus/mm
2
was therefore determined by counting total number of glomeruli in kidney sections from pups starting from P0 to P20. The salt-resistant 26-P congenic strain exhibited significantly greater nephron density 9.03 and 7.07 glo/mm
2
compared to 6.89 and 4.85 glo/mm
2
in SS rat at day P15 and P20, respectively. It appears that the Brown Norway pappa2 allele variant prevents the reduced nephron numbers observed in SS rats and thereby protects these congenic rats from salt-induced hypertension.
Abstract 100: Hip and Pelvic Fracture Risk in Adults Treated with Three Different Classes of Antihypertensive Medications: The ALLHAT Study
Authors: Oparil Suzanne , Puttnam Rachel , Davis Barry , Pressel Sara , Whelton Paul , Cushman William , Louis Gail , Margolis Karen , Williamson Jeff , Ghosh Alokananda , Einhorn Paula , Barzilay Joshua
Published: June 2021

Observational studies suggest that thiazide-type diuretics reduce fracture risk compared to other antihypertensive medications. The effects of calcium channel blockers (CCB) and angiotensin converting-enzyme inhibitors (ACEi) on fracture risk have not been well studied. We examined the relationship of antihypertensive drug therapy and hip and pelvic fracture hospitalizations in the Antihypertensive and Lipid Lowering Treatment to Prevent Heart Attack

Trial

(

ALLHAT

). It included >33,000 participants randomized to the thiazide-type diuretic chlorthalidone (C), ACEi lisinopril (L), or CCB amlodipine (A) as first line hypertension (HTN) therapy. Mean follow up was 4.9 years during the randomized phase (in-trial), and 5 additional years after the conclusion of the trial (post-trial) using linkage to national data bases. Risks of hip and pelvic fractures for L and A relative to C were derived from Cox models. There were 341 hip and pelvic fractures in-trial.

Participants assigned C had the lowest risk and those assigned L the highest (
Figure 1a).
The adjusted risk for L compared to C was 1.33 (95% CI 1.02-1.73; p=.04). Participants assigned A had intermediate risk compared to C (HR 1.22, 95% CI 0.93-1.59). During the combined in-trial and post-trial periods (
Figure 1b
), there were 646 fractures; the results were similar to the in-trial results, although differences were not statistically significant. Participants randomized to C continued to have the lowest risk of fractures after in-trial period, suggesting a legacy effect from prior C use. These findings have public health importance given the high prevalence of HTN in older adults and the widespread use of A and L in older adults.
Abstract 114: Renal Denervation Reduces Monocyte Activation: An Anti-inflammatory Effect Relevant for Cardiovascular Risk
Authors: Zaldivia Maria Teresa Kristina , Rivera Jennifer , Hering Dagmara , Marusic Petra , Marusic Petra , Sata Yusuke , Eikelis Nina , Lee Rebecca , Lambert Gavin W. , Htun Nay M. , Duval Jacqueline , Hammond Louise , Eisenhardt Steffen , Flierl Ulrike , Schlaich Markus , Peter Karlheinz
Published: June 2021

Background:
Over-activation of renal sympathetic nervous system and low-grade systemic inflammation are thought to be common features of hypertension. Renal Denervation (RDN) reduces sympathetic activity in patients with resistant hypertension. However, its effect on systemic inflammation has not been investigated.

Aim:
To determine the effect of RDN-induced sympathetic inhibition on monocyte activation and systemic inflammation in hypertensive patients.

Methods:
Peripheral blood was obtained from 42 patients who underwent RDN for uncontrolled blood pressure (BP) at baseline, at 3 months and 6 months post-procedure. Ambulatory BP, overall activation status of monocyte as well as monocyte subsets and inflammatory markers were assessed at each time point.

Results:
RDN significantly lowered 24-hour ambulatory BP at 3 months (150.5/81.0 mmHg to 144.7/77.9 mmHg), which was sustained at 6 months (144.7/78.6 mmHg). The overall monocyte activation was significantly decreased (3 months, 4079.4 MFI to 3182.0 MFI; 6 months, 3457.62 MFI) post-RDN, specifically in the subset of classical monocytes (6 months, 4696.8 MFI to 3958.8 MFI). In line with this, reduction of several inflammatory markers were observed, including monocyte-platelet aggregates at 3 months (34% [680 of 2000 monocyte events] to 11.85% [237 of 2000 monocyte events]) and plasma levels of MCP-1 (3 months, 144.9 pg/ml to 100.1 pg/ml; 6 months, 122.2 pg/ml), IL-1β (3 months, 18.3 pg/ml to 10.8 pg/ml; 6 months, 12.2 pg/ml), TNF-α (3 months, 167.5 pg/ml to 78.4 pg/ml; 6 months, 111.1 pg/ml), IL-12 (3 months, 59.8 pg/ml to 9.9 pg/ml; 6 months, 21.4 pg/ml) and IL-6 (3 months, 2.4 pg/ml to 1.5pg/ml; 6 months, 1.9 pg/ml). A positive correlation was observed between baseline muscle sympathetic nerve activity and monocyte activation (R=0.62) and changes observed at both time points (3 months, R=0.63; 6 months, R=0.88) post-procedure.

Conclusions:
Inhibition of sympathetic activity via RDN is associated with a reduction of monocyte activation and other circulating inflammatory markers in hypertensive patients. These findings point to a direct interaction between the inflammatory and sympathetic nervous system, which is of central relevance for the understanding of beneficial cardiovascular effects of RDN.
Abstract 115: Blood Pressure Trajectories In The Acute Phase And Major Clinical Outcomes Among Patients With Ischemic Stroke
Authors: Li Changwei , Zhang Yonghong , Chen Jing , Xu Tan , Wang Dali , Chen Chung-Shiuan , Sun Yingxian , Peng Hao , He Jiang
Published: June 2021

We studied the relationship of blood pressure (BP) trajectories during the first seven days after symptom onset with short- and long-term major clinical outcomes among patients with acute ischemic stroke. A total of 4,036 patients with acute ischemic stroke and elevated systolic BP from the CATIS trial were included in this analysis. Three BPs were measured every 2 hours for the first 24 hours, every 4 hours during the second and third days, and every 8 hours thereafter for the remainder of the seven days. Latent class models were used to identify subgroups that share a similar underlying trajectory in BP in the acute phase. Five systolic BP trajectories of high, high-to-moderate low, moderate high, moderate low, and low were identified. Compared to the high trajectory, multiple-adjusted odds ratios (95% CI) of death and major disability at 3 months for high-to-moderate low, moderate high, moderate low, and low trajectories were 0.61 (0.44 to 0.86), 0.63 (0.48 to 0.84), 0.49 (0.37 to 0.65), and 0.42 (0.30 to 0.59), respectively (overall p<0.0001). Likewise, the corresponding multiple-adjusted odds ratios at 2 years were 0.64 (0.46 to 0.90), 0.78 (0.59 to 1.04), 0.49 (0.37 to 0.66), and 0.49 (0.34 to 0.69), respectively (overall p<0.0001). These data indicate that individuals with a consistently high systolic BP during the acute phase of ischemic stroke had the highest risk of short- and long-term death and major disability. In addition, moderate systolic BP reduction to below 140 mmHg from higher levels lowers risk of short- and long-term death and major disability.
Abstract 127: Enhanced Endothelial Mineralocorticoid Receptor Signaling Prompts Vascular Inflammation and Stiffness Through MiRNA-103-mediated Suppression of KLF4
Authors: Jia Guanghong , Habibi Javad , Aroor Annayya R. , Sun Zhe , DeMarco Vincent G , Meininger Gerald A , Jaffe Iris Z , Sowers James R
Published: June 2021

Enhanced activation of mineralocorticoid receptors (MRs) impairs insulin metabolic signaling, increases oxidative stress, and induces inflammation with associated cardiovascular abnormalities. Our previous data in female mice suggests that activation of endothelial cell MRs (ECMR) contributes to development of vascular stiffness partly by impairing insulin metabolic signaling and reducing endothelial derived nitic oxide (NO) production. Emerging information suggests that microRNA 103 (miR103) is upregulated and thus promotes endothelial inflammation and atherosclerosis in both ob/ob mice and western diet-induced obese C57BL/6J mice. However, the interaction of ECMR and miR103 in the promotion of vascular inflammation and stiffness has not been explored. We hypothesized that ECMR signaling prompts vascular inflammation and stiffness through miRNA-103-mediated suppression of Kruppel-like factor-4 (KLF4). Female ECMR knockout (ECMR
-/-
) and wild-type littermate females were treated with aldosterone (Aldo) (250 μg/kg/day) via osmotic minipumps for 4 weeks. Aldo infusion induced endothelium stiffness and impaired aortic relaxation in wild-type mice as determined by ex vivo atomic force microscopy and wire myograph techniques, respectively. The elevated aortic stiffness and impaired relaxation was accompanied by increases in expression of miR103 and intercellular adhesion molecule 1 (ICAM-1) and a reduction in phosphorylation of serine (Ser) 1177/activation of endothelial NO synthase (eNOS). These Aldo-induced endothelial abnormalities were prevented in ECMR
-/-
mice. Furthermore, application of a miR103 inhibitor to ECs in vitro attenuated Aldo (10
8
M)-induced a decrease in KLF4 expression, which has anti-inflammatory functions mediated by upregulation of phosphorylation of eNOS and downregulation of ICAM-1. These findings suggest that increased ECMR signaling and associated miR103 activation plays a key role in Aldo-induced KLF4 suppression and associated vascular inflammation and aortic stiffness in females.
Abstract 128: Leptin: A Regulator of Aldosterone Synthase Expression & Aldosterone Secretion in Visceral Adipocytes, in Mice
Authors: Belin de Chantemele Eric J , Huby Anne-Cecile , Menk P. T , Chen Weiqin , Lane Brian , Antonova Galina
Published: June 2021

Obesity is associated with inappropriately high aldosterone levels, which contribute to the development of metabolic and cardiovascular disorders. The origin of these high aldosterone levels is incompletely understood. We recently demonstrated that the adipocyte-derived hormone leptin regulates aldosterone synthase (CYP11B2) expression and stimulates aldosterone release from adrenal zona glomerulosa cells. Recent studies demonstrate that adipocytes express CYP11B2 and secrete aldosterone. However, the mechanisms regulating aldosterone release from adipocytes remain unclear. Likewise, whether visceral (Visc) and subcutaneous (SubQ) adipose tissue contribute to a similar extent to aldosterone production is unknown. We tested the hypothesis that leptin increases adipocyte CYP11B2 expression and aldosterone production and investigated whether Visc and SubQ adipose tissues respond similarly to leptin. Immunostaining of mouse adipose tissue cross-sections and isolated mature adipocytes revealed that Visc and SubQ adipose tissue express leptin receptors. Treatment of mouse freshly isolated mature adipocytes, non-differenciated (stromal fraction) and differentiated adipocytes revealed that leptin dose-dependently increased CYP11B2 expression and aldosterone production in Visc adipose tissue only. Although leptin receptor and CYP11B2 levels were similar in SubQ and Visc adipocytes, SubQ adipocytes were unresponsive to leptin. The physiological relevance of these in vitro data was tested by measuring plasma aldosterone levels in mice deprived of adipose tissue (lipodystrophic mice) treated with leptin. Absence of adipose tissue in lipodystrophic mice blunted leptin-induced increases in aldosterone levels (WT-vehicle: 471±82 vs. WT-Leptin: 1699±396, p<0.05; KO-vehicle: 539±71 vs. KO+leptin: 787±156, NS). The human relevance of these data was determined by reporting that CYP11B2 expression gradually increased with body mass index in human mediastinal and omental fat depots. In summary these data strongly suggest that leptin regulates CYP11B2 levels and aldosterone release in visceral adipose tissue and that leptin-induced, adipocyte-derived aldosterone may contribute to obesity-associated hyperaldosteronism.
Abstract 135: Obesity Accelerates the Deterioration of Renal Function in Developmental Programming of Hypertension. Role of Angiotensin Ii and Oxidative Stress
Authors: Tapia Antonio , Moreno Juan M , Llinas Maria T , Salazar F. Javier
Published: June 2021

Numerous studies have shown gender-dependent differences in the deterioration of renal function in models of developmental programming of hypertension (DPH). It is also known that obesity is associated to changes in renal function and that both angiotensin II (Ang II) and oxidative stress are involved in the renal alterations that occur in obesity and in animals with DPH. The main objectives were to examine whether the increment of arterial pressure (AP) and the deterioration of renal function are accelerated as a consequence of obesity in SD rats with DPH; whether these changes are gender-dependent; and to evaluate the role of Ang II and oxidative stress in these AP and renal function changes. A high fat diet (60%) was given during the first 4 months of age and DPH was induced by an AT receptor antagonist during nephrogenic period (ARAnp). Systolic AP (mmHg) was greater (P<0.05) in ARAnp-obese rats (167 ± 3 in ♂; 146 ± 4 in ♀) than in ARAnp (155 ± 3 in ♂; 137 ± 3 in ♀); obese (147 ± 2 in ♂; 137 ± 2 in ♀) or control (127 ± 1 in ♂; 119 ± 2 in ♀) rats. Three days administration of candesartan (7 mg/kg/day) led to a decrease in AP that was greater (P<0.05) in ARAnp-obese rats (55 ± 3 in ♂; 45 ± 4 in ♀) than in ARAnp (40 ± 3 in ♂; 37 ± 4 in ♀); obese (38 ± 4 in ♂; 27 ± 4 in ♀) or control (12 ± 2 in ♂; 14 ± 3 in ♀) rats. The acute Ang II infusion (30 ng/kg/min) induced an increase in renal vascular resistance (mmHg/ml/min/gr kw) that was also greater in ARAnp-obese rats (217 ± 45% in ♂; 145 ± 38% in ♀) than in ARAnp (103 ± 9% in ♂; 97 ± 8% in ♀); obese (106 ± 14% in ♂; 106 ± 17 in ♀) or control (51 ± 7% in ♂; 51 ± 10% in ♀) rats. The response to candesartan or Ang II infusion in ARAnp-obese rats was gender-dependent and may be explained by an enhanced oxidative stress. The expression of P67phox in the renal cortex was greater (P<0.05) in ARAnp-obese rats (3,00 ± 0,05 in ♂; 2,60 ± 0,04 in ♀) than in ARAnp (1,16 ± 0,04 in ♂; 1,66 ± 0,03 in ♀); obese (0,94 ± 0,06 in ♂; 1,02 ± 0,02 in ♀) or control (1,00 ± 0,02 in ♂; 1,02 ± 0,023 in ♀) rats. The results of this study suggest that obesity at an early age enhances the hypertension and accelerates the deterioration of renal function that occurs when cardiovascular disease is programmed during the perinatal period. It is also shown that Ang II and oxidative stress seems to play an important role in these AP and renal function changes.
Abstract 136: Influence of Race and Obesity on the Renin-Angiotensin-Aldosterone System in Adolescents Born Preterm
Authors: South Andrew M , Nixon Patricia A , Chappell Mark C , Diz Debra I , Russell Gregory B , Snively Beverly M , Shaltout Hossam A , Rose James C , O’Shea T. M , Washburn Lisa K
Published: June 2021

Background:
While neonatal morbidity and mortality in preterm infants have improved dramatically, the long-term cardiovascular and renal consequences of prematurity are incompletely understood. Prematurity may induce programmed changes in the renin-angiotensin (Ang)-aldosterone system (RAAS), a key regulator of cardiovascular and renal function. Race and obesity influence the RAAS and may modify the effects of prematurity on the RAAS. We hypothesized that the RAAS differs by race and obesity in adolescents born prematurely with very low birth weight (VLBW).

Methods:
A cohort of 173 adolescents with VLBW was evaluated at age 14. We measured renin, aldosterone, Ang II, and Ang-(1-7) in the plasma; Ang II, Ang-(1-7), and creatinine in the urine; and we calculated the aldosterone/renin ratio and the Ang II/Ang-(1-7) ratios. We used general linear regression models to estimate the difference in the RAAS according to overweight/obesity (body mass index ≥85% for age and sex) and race, adjusting for confounding variables.

Results:
On unadjusted analyses as well as analyses adjusted for sex, antenatal corticosteroid exposure, and maternal hypertension, black race was associated with decreased urinary Ang-(1-7)/creatinine (adjusted estimate -0.18, 95% CI -0.36 to -0.01,
p
= 0.04) and decreased renin (-0.36, -0.68 to -0.05,
p
= 0.03). In analyses stratified by sex, black males, but not black females, had decreased renin (-0.63, -1.1 to -0.16,
p
= 0.01) and aldosterone (-0.61, -1.19 to -0.04,
p
= 0.04). Obesity was associated with increased urinary Ang II/(1-7) (0.29, 0.04 to 0.53,
p
= 0.02), decreased plasma Ang-(1-7) (-0.4, -0.8 to -0.002,
p
= 0.05), increased plasma Ang II (0.21, 0.03 to 0.39,
p
= 0.02), and increased plasma Ang II/(1-7) (0.61, 0.2 to 1.01,
p
= 0.004).

Conclusions:
In adolescents born with VLBW, there is racial variation in the RAAS. Black adolescents, especially males, have an altered renal RAAS and lower renin and aldosterone. Obesity is associated with a potentially deleterious alteration in the RAAS, with a shift in the renal and systemic RAAS towards Ang II and away from Ang-(1-7). These shifts in the RAAS associated with race and obesity may increase the risk of renal and cardiovascular disease in adolescents born with VLBW.
Abstract P100: Interaction of the Mineralocorticoid Receptor with RACK1
Authors: Kuppusamy Maniselvan , Gomez-Sanchez Elise P , Gomez-Sanchez Celso E
Published: June 2021

The mineralocorticoid receptor (MR) is a multifunctional ligand dependent transcription factor in the steroid receptor superfamily. The MR mediates aldosterone regulation of electrolytes and blood pressure. MR transcriptional co-regulators to influence specific gene transcription. We used a yeast two-hybrid system to find proteins that interact with a full-length MR as bait.

Results:
Among other proteins, we found a specific interaction of MR with RACK1 (Receptor for Activated C Kinase 1), a scaffolding protein. Overexpression of RACK1 using a tetracycline-inducible lentivirus in mouse cortical collecting duct M1 cells stably expressing a reporter Gaussia luciferase gene under a hormone-response element promoter resulted in enhanced agonist-dependent MR transactivation. Inhibition of RACK1 protein expression by short hairpin RNA led to a significant reduction in MR activation of the reporter gene. RACK1 regulation of MR action is mediated through the PKC-β signaling pathway. MR and RACK1 co-precipitated using a MR antibody in Sprague-Dawley brain tissue and M1 cells and immunofluorescent histochemistry showed MR and RACK1 co-localization in Male Sprague-Dawley brains and M1 cells.

Conclusion:
The scaffolding protein RACK1 is associated with MR under basal and agonist stimulated conditions and facilitates agonist stimulated MR actions through PKC-β. These findings indicate that RACK1 function as a novel co-activator of MR.
Abstract P102: Differences in Angiotensin-(1-7)/alamandine-mediated Signaling in Tumoral and Normal Cell Lines
Authors: Sampaio Walkyria O , Oliveira Marilene Luzia , Silva Felipe A , Silva Leticia B , Etelvino Gisele M , Passos-Silva Danielle G , Santos Robson A
Published: June 2021

Previously studies have demonstrated that besides its actions in the cardiovascular system, Angiotensin-(1-7) also plays a role in inhibiting tumoral growth. The role of recently described Alamandine in this field is not clear. The signaling pathways underling anti-tumoral actions of these peptides are also poorly understood. Therefore, the aim of this study was to elucidate the modulatory effect of Ang-(1-7) and Alamandine in the PI3K cascade, a well-known signaling pathway described to be involved in proliferation and cancer. To achieve this goal, we stimulate human pancreatic and lung cancer cell lineage (Miapaca and A549), as well as a control cell lineage (VERO) with Ang-(1-7) and Alamandine. Through western blotting analysis, our data suggest that both Ang-(1-7) and Alamandine activate the phosphatase pTEN (dephosphorylation of S380/T382/T383) (48±4% after 24 hours Ang-(1-7) treatment in miapaca and A549 in comparison of non-treated cells, p<0.05) (60±5 and 48±4% of phosphorylation level after 24 hours Alamandine treatment in miapaca and A549, respectively, in comparison of non-treated cells, p<0.05), which dephosphorylates PI3K, inactivating this kinase. Furthermore, AKT phosphorylation is transient, followed by a significant dephosphorylation when compared to the non-treated cells (30±5% after 24 hours Ang-(1-7) treatment in miapaca in comparison of non-treated cells, p<0.05). Ang-(1-7) also inhibits a PTEN downstream effector kinase, mTOR through dephosphorylation of T246 (70±5% after 24 hours Ang-(1-7) treatment in miapaca in comparison of non-treated cells, p<0.05). These effects were not observed in control non-tumoral cells (VERO cells). As previously demonstrated with Ang-(1-7) stimulation, Alamandine also induces the FOXO1 activation and migration to the nucleus in A549 (122 ± 8 of A.U. of fluorescence at 4 hours after alamandine treatment vs 46±4 A.U. at control, p<0.001) and Miapaca cells (67 ± 5 of A.U. of fluorescence at 4 hours after alamandine treatment vs 16±2 A.U. at control, p<0.001). These results indicate that, in contrast to normal tissues, Ang-(1-7) and Alamandine decreases, through PTEN activation, PI3K/AKT pathway in tumoral cells.
Abstract P104: Role of Adenosine Receptors in Angiotensin II Dependent Hypertension in Mice
Authors: Yadav Vishal R , Zhou Zhichao , Teng Bunyen , Mustafa S. Jamal
Published: June 2021

Our recent finding showed that acute angiotensin (ANG) II stimulation enhanced A
1
adenosine receptor (AR) dependent cytochrome P450 (CYP) 4A mediated contraction in mouse mesenteric arteries (MA). In addition, A
2B
AR dependent and partly K
ATP
channel mediated relaxation was also reduced with ANGII stimulation. These data suggest a possible interaction between ANG II and ARs at vascular level which may play a role in ANG II dependent hypertension.

We developed ANG II (1000 ng/kg/min, 21 days) infused hypertension mouse model and tail cuff was used to measure blood pressure. Aorta as a conduit and MA (second order) as a resistance artery were chosen for muscle tension studies. Protein expressions in aortas and MAs were analyzed by western blot.

Infusion of ANG II increased systolic arterial blood pressure (SAP) in mice. Particularly, in ANG II infused mice, SAP (in mmHg, mean ± SD) at day 0 was 101.6 ± 6.675 which increased to 156.7 ± 15.68 on day 21 (p<0.05) while in saline control mice, SAP was 100.8 ± 6.753 (day 0) and changed to 105.4 ± 7.620 (day 21).

Higher expression of A
1
AR (~155% over 100%) and CYP4A (~50%, ~0.6 over 0.3, ratio to actin) was present in MAs in hypertensive compared to control mice. Expression of A
1
AR and CYP4A was comparable in aortas of hypertensive and control mice.

CCPA (A
1
AR agonist) -induced concentration dependent contraction (% normalize to phenylephrine contraction) was significantly reduced (~15% from 40% at 100 nM) in aorta of hypertensive mice compared to control. NECA (non-selective AR agonist) produced a significantly higher (~ -15% from 0% at 10 μM) relaxation in aorta of hypertensive mice. In MAs, CCPA-induced contraction was reduced in hypertensive mice. NECA induced relaxation was comparable in MAs. Pinacidil (K
ATP
channel opener) induced relaxation was significantly lower (~0% from -25% at 100 nM) in MAs of hypertensive mice.

In conclusion, our data reveal important role for A
1
AR dependent signaling in ANG II induced hypertension. Higher A1AR and CYP4A may reduce K
ATP
channel dependent relaxation of MAs aiding vascular contraction and blood pressure. Further studies involving ARs pharmacological inhibitors and genetic models are required to fully understand the relationship between ANG II and ARs in the development of hypertension.
Abstract P107: Hypertension Enhances the Differentiation of Cardiac Fibroblasts Into Myofibroblasts After TGF-beta1 Treatment
Authors: Perrucci Gianluca L , Corlian!ò Maria , Tosi Delfina , Nigro Patrizia , Bulfamante Gaetano , Pompilio Giulio , Lombardi Federico
Published: June 2021

Objectives:
In cardiac fibrosis associated with hypertension, TGF-beta1 plays a key role by acting on differentiation of cardiac fibroblasts (CF) into alpha-smooth muscle actin (alpha-SMA)-positive myofibroblasts. In this study, we tested the effect of TGF-beta1 during the myofibroblast differentiation process of CF from normotensive and hypertensive rats.

Methods:
CF were obtained by enzymatic digestion of hearts isolated from Spontaneously Hypertensive (hCF) and normotensive Wistar Kyoto (nCF) rats (n=5 rat/group). Gene and protein expression in CF was evaluated by Western blot and qRT-PCR analyses, respectively. Immunohistochemistry analysis for integrin alpha-v beta-5 was performed on rat cardiac tissue (n=5 rat/group).

Results:
Cultured hCF showed an enhanced SMAD2/3 activation and alpha-SMA protein expression after treatment with TGF-beta1 (5 ng/ml) in comparison with nCF. Alpha-SMA up-regulation was further confirmed by qRT-PCR analysis that showed a significant increase in alpha-SMA gene expression in hCF after TGF-beta1 treatment (2.78±0.25 vs 2.01±0.21 fold increase,
p
<0.05). Moreover, immunostaining on cardiac tissues revealed a higher expression of integrin alpha-v beta-5 in hypertensive
vs
normotensive rat hearts (345.3±170.0 vs 48.2±22.3 mm
2
of integrin-positive area,
p
<0.05). This result was also confirmed
in vitro
; indeed, integrin alpha-v beta-5 gene expression in hCF increased 2.8-fold in basal condition and 5.12-fold after TGF-beta1 treatment when compared to untreated nCF.

Conclusions:
Taken together, these results suggest that hCF are more prone to upregulate integrin alpha-v beta-5 and consequently differentiate into myofibroblasts
in vitro
under TGF-beta1 treatment. Thus, targeting alpha-v beta-5 might open a novel prospective for the treatment of fibrosis in hypertensive hearts likely reducing integrin-mediated TGF-beta1 activation.
Abstract P116: Exercise Ameliorated Cognitive Impairment Through ER Stress Mitigation in Alcohol Treated Exercise Ameliorated Cognitive Impairment Through Er Stress Mitigation in Alcohol Treated C57bl/6 Male Mice
Authors: George Akash , Richardson Kennedy , Zhai Yuankun , Tyagi Suresh C , Tyagi Neetu
Published: June 2021

Alcohol consumption is a potent inducer of oxidative stress (OS). Oxidative stress cause disturbance of endoplasmic reticulum (ER) homeostasis, that triggers ER stress (ERS), cause neuronal damage in the brain. Our Previous data indicate that Alcohol consumption induces mitochondrial dysfunction and free radical production in mouse cerebral cortex. Exercise has been recommended by clinicians as a secondary protective therapy; however, its effect on brain functions through ER stress has not been fully explored. Therefore, we hypothesized that exercise improves Alcohol-induced neurodegeneration and decline in cognitive function through ER stress mitigation. To test this hypothesis, we selected 10-12 weeks old male wild-type mice (C57BL/6, WT), grouped as follows: 1) WT, 2) WT+ Alcohol, 3) WT+ Exercise, 4) WT+ Alcohol + Exercise. Mice were given an intraperitoneal injection of Alcohol (1.5g/kg BW) or saline solution every day for 8 weeks. The mice were exercised for 8 weeks on a treadmill with a controlled speed of 7 meters/min for the first week, the speed of 10 meters/min for the second week and 11 meters/min in the following weeks and a total of 330 meters every day. After each 110 meters mice were given rest of 10 minutes. Cognitive and behavior alterations were assessed by novel object recognition, Passive avoidance, and Y-maze tests. Our result showed there is a significantly impaired cognitive and behavior functions (600.00 ± 0.00 vs 480 ± 20.00, P<0.05) in Alcohol-treated group compared to WT control mice. However exercised significantly improved (0.37 ± 0.05 vs 0.63 ± 0.04 P<0.01) these functions as compared to Alcohol-treated group. Also, we observed an elevated blood pressure in the Alcohol-treated group (123.50 ± 1.17) and exercise brought that to the normal level (108.98 ± 4.47, P<0.01). In addition, the effect of exercise on neuronal survival in the Alcohol-treated mouse brain was confirmed by a decrease in by fluoro-jade C reactivity. Taken together, our results indicate a myriad of beneficiary effects of exercise over ER mitigation in Alcohol-treated mice. Furthermore, our findings suggest exercise alleviates neurodegeneration and cognitive dysfunction and thereby improving total brain function. This work was supported by NIH grant HL107640-NT
Abstract P120: Exercise in Patients With Chronic Kidney Disease on Hemodialysis: Systematic Review and Network Meta-analysis of Randomized Clinical Trials
Authors: Scapini Kátia B , Moraes Oscar A , Sbruzzi Graciele , Inácio José F , Rodrigues Clarissa G , Leguisamo Camila P , Tourinho Filho Hugo , Böhlke Maristela , Irigoyen Maria C
Published: June 2021

Background:
Exercise provides beneficial effects in ESRD, however few studies compare efficacy of different modalities of exercise in this population. To overcome the restrictions of limited available comparisons we employed network meta-analysis of randomized clinical trials (RCTs) to compare the efficacy of different modalities of exercise on aerobic capacity, arterial blood pressure and hemodialysis efficacy in ESRD adults patients on hemodialysis treatment.

Methods and Findings:
We searched PubMed, EMBASE, COCHRANE CENTRAL, WEB OF SCIENCE and LILACS databases for RTCs with adults ESRD patients on hemodialysis that compared aerobic exercise (AE), resistance training (RT) or combined training (CT) with a control group. Were included 28 trials involving1045 ESRD patients on hemodialysis. Direct meta-analysis show that both, AE (3.35, 95% CI: 1.79, 4.91 ml/kg/min) and CT (5.0, 95% CI: 3.50, 6.50 ml/kg/min) improved aerobic capacity while only CT reduced systolic (-5.88, 95% CI: -9.83, -1.93 mmHg) and diastolic (-3.93, 95% CI: -6.23, -1.60 mmHg) arterial blood pressure. Regarding hemodialysis efficacy, only AE was able to improve Kt/V (0.16, 0.05, 0.26). Network meta-analysis showed that AE and CT was superior to control group for aerobic capacity and CT presented 92.8% of probability to be ranked as first most effective treatment. For arterial blood pressure, only CT was superior to control group and for diastolic blood pressure combined training was also significantly superior to AE. Moreover, CT presented 94.7% probability of being best treatment for systolic arterial pressure and 98.7% for diastolic arterial pressure. None modality of exercise was superior to control treatment for hemodialysis efficacy.

Conclusions:
The main contribution of this network meta-analysis is to ranking the potential benefits of different exercise modalities on health outcomes in hemodialysis patients. The analysis found that combined training, aerobic plus resistance training, is the most effective modality to increase aerobic capacity, and blood pressure control, in hemodialysis patients. This finding comes to fill the gap created by the lack of head-to-head comparison of different modalities of exercise in chronic kidney disease patients.
Abstract P128: Enhanced NOS1β in the Macula Densa Contributes to the Diabetic Hyperfiltration
Authors: Wei Jin , Zhang Jie , Zhang Gensheng , Wang Lei , Wang Shaohui , Liu Ruisheng , Buggs Jacentha
Published: June 2021

Hyperfiltration is common in early diabetes and considered a risk factor for diabetic nephropathy. Inhibited tubuloglomerular feedback (TGF) mediated by less NaCl delivery at the macula densa contributes to the diabetic hyperfiltration. Nitric oxide (NO) released from macula densa via neuronal nitric oxide synthase (NOS1) inhibits TGF response. We recently demonstrated the significance of TGF response in the long-term control of GFR, sodium excretion and blood pressure, mediated by macula densa NOS1β. However, whether the NOS1β-mediated TGF response play an important role in the diabetic hyperfiltration is unknown. We hypothesized that macula densa NOS1β is upregulated in diabetes, which blunts TGF response and contributes to the diabetic hyperfiltration. The mice with deletion of NOS1 specifically from the macula densa (MD-NOS1KO) and wild-type C57BL/6 (WT) mice were used. Diabetes was induced by alloxan (55 mg/kg i.v.) with blood glucose from 350 to 450 mg/dl. Expression of NOS1 splice variants was measured with real-time PCR and Western blot. GFR was measured by plasma FITC-inulin clearance following a single bolus intravenous injection in conscious mice. GFR increased by 24.7% in diabetic WT mice (from 241.60±19.73 to 301.35±21.76 μl/min) and only by 16.5% in diabetic MD-NOS1KO mice (from 236.61±16.12 to 275.61±11.73 μl/min) (n=6/group, p<0.01 vs baseline; p<0.05 vs WT). To determine whether glucose induces hyperfiltration mediated by macula densa NOS1, intravenous infusion of 2 μl/g glucose solution (2 M) into non-diabetic C57BL/6 mice elevated blood glucose to about 350 mg/dl and increased GFR by 19.1% (from 236±15.4 to 281±9.7 μl/min, n=6, p<0.05), but did not significantly increase GFR (from 223±6.9 to 240±15.7 μl/min) in non-diabetic MD-NOS1KO mice (n=6, p<0.01 vs WT). The expression of NOS1β was upregulated by 8.9±1.3 folds in protein level and 10.1±2.1 folds in mRNA level in diabetic WT mice (n=4, p<0.01). Present study provided a novel mechanism for diabetic hyperfiltration mediated by macula densa NOS1β. Enhanced NOS1β in the macula densa contributes to the diabetic hyperfiltration.
Abstract P132: Role of NO/NOS System in Blood Pressure Regulation in Postmenopausal Female SHR with Hormone Replacement Therapy
Authors: Dalmasso Carolina , Maranon Rodrigo O , Patil Chetan N , Yanes-Cardozo Licy L , Reckelhoff Jane F
Published: June 2021

Estradiol (E
2
) and testosterone (T) stimulate synthesis of nitric oxide (NO). Since the WHI study, postmenopausal women are given E
2
and/or T supplements to alleviate the symptoms of menopause. NO is a key regulator of blood pressure (BP), mediating endothelial and vascular function. Studies from our lab showed that E
2
plays little role in BP control in young female SHR; however, whether E
2
replacement with or without T would reduce BP in postmenopausal female SHR is not known. If E
2
does not reduce BP in old female SHR, it is possible that there is a defect in the NO system that prevents the vasodilatory E
2
effect mediated by NO. The hypothesis tested in this study was that E
2
/T replacement would reduce BP in old female SHR, and if not, the mechanism responsible is a deficient NO system that is incapable of upregulating in response to E
2
. After baseline (B) mean arterial pressure (MAP; telemetry), female SHR (19 mos, n=5) were implanted with 17-β E
2
(0.1 mg/pellet) and T (5mm in silastic tubes) and MAP was measured. After a transient reduction in MAP over 2-3 days (B: 175±5; E
2
+T: 161±4 mm Hg, p<0.05), MAP returned to baseline levels by day 4 (176±5 mm Hg). These data suggest that the acute vasodilatory response to E
2
/T in old female SHR was intact. On day 8 T tubes were removed, and MAP was measured for additional 16 days. Removal of T had no effect on BP (175±5 mm Hg). To evaluate NO system, rats were given: 1) 2% L-Arginine (L-Arg, 21 d); 2) 0.1% sodium nitrite (NaNO
2
, 6 d); 3) nitro-L-arginine methyl ester (L-NAME, 4mg/kg/d, 5 d). L-Arg supplement failed to change MAP (B: 175±5, L-Arg: 176±5 mm Hg, p<NS). In contrast, NaNO
2
did decrease MAP (B: 176±5, NaNO
2
: 161±3 mm Hg, p<0.05), suggesting a deficient endogenous synthesis of NO but the ability of the old female SHR to respond to an NO donor. L-NAME increased MAP (B: 176±5; L-NAME: 189±3 mm Hg, p<0.05). In total, these data suggest that the NO system in old female SHR is capable of responding appropriately to NO donors or complete blockade. However, the lack of response to L-Arg suggests a deficiency in the ability to normally synthesize NO, and thus may in part be responsible for the lack of a depressor response to E
2
, and therefore, may contribute to the elevated BP in old female SHR. Supported by NIH-R01HL66072, PO1HL51971 (JFR), 14POST18640015 (ROM).
Abstract P140: Epigenetic Regulation of Nox4 in Hypertension-induced Injury in Aged Kidney
Authors: Ren Lu , Das Purkayastha Biswa , Sen Utpal , Pushpakumar Sathnur
Published: June 2021

The prevalence of hypertension increases with age. At the cellular level, oxidative stress is a major contributing factor to the pathogenesis of hypertension-induced kidney damage. The nicotinamide adenine dinucleotide phosphate (NADPH) family is one of the major sources of reactive oxygen species generation in the body. Although the isoform, NADPH oxidase 4 (Nox4) is highly expressed in the kidney, its role in kidney diseases remains controversial as both pathogenic and protective effects have been described. In addition, its role in hypertension-induced kidney damage in aging remains unexplored. Current evidence supports the involvement of epigenetics in oxidative stress. The aim of the present study was to investigate the role of Nox4 in Ang-II induced kidney damage in aged mice and the potential role for epigenetic regulation. Wild type (WT, C57BL/6J) mice aged 12-14 wk and 75-78 wk were used in this study and treated with Ang-II (1000 ng/kg/min) for 4 weeks. Control mice received saline infusion. Blood pressure (BP) was measured twice weekly. Aged mice exhibited higher mean BP than young mice after Ang-II treatment (135.76±4.8 vs. 118.7±12.28 mmHg) and decreased renal vascular density on barium angiography. Dihydroethidium staining revealed increased oxidative stress in hypertensive kidney of aged mice. In the aged kidney, protein expression of mitochondrial antioxidant enzymes, manganese superoxide dismutase and catalase was decreased by > 2-fold compared to young, and decreased further after Ang-II infusion. The renal sensors of energy and redox state, SIRT1 and SIRT3, showed similar decrease in aged kidney compared to young kidney in response to Ang-II. Nox4 and p22
phox
protein expression was upregulated in the hypertensive kidneys of aged mice. The epigenetic mechanism involving DNA methylation showed increased DNA methyltransferases, DNMT1, 3a and 3b in the aged kidney. Following Ang-II, the aged kidney showed further increase of DNMT1 and 3a and a decrease in DNMT3b. Conclusion: Endogenous expression of Nox4 is upregulated in hypertensive kidney and is associated with alteration of DNA methyltransferases suggesting epigenetic regulation of oxidative stress in aged mice.
Abstract P145: Renal Inflammation and Injury is Associated with Increased Lymphangiogenesis in Hypertension
Authors: Kneedler Sterling C , Phillips Lauren , Hudson Kayla R , Beckman Katharine M , Parrish Alan R , Doris Peter A , Mitchell Brett M
Published: June 2021

Hypertension is associated with immune system activation and inflammation. Renal infiltration of both innate and adaptive immune cells contributes to injury, dysfunction, and increased blood pressure. Activated immune cells that exit blood vessels into the interstitium then travel through lymphatic vessels to draining lymph nodes where they signal to other immune cells to increase the immune response. It is unknown how renal lymphatic vessels change in the context of hypertension, immune system activation, inflammation, and injury. We hypothesized that renal macrophage infiltration, inflammation, and injury would significantly increase lymphangiogenesis in various strains of rats. SHR rats that exhibit hypertension and renal injury (SHR-A3 strain) had significantly increased numbers of renal lymphatic vessels at 40 weeks of age compared to WKY controls (total of 3 fields of view: 52 ± 1 vs. 28 ± 1; p<0.05). This was associated with increased renal macrophage infiltration. SHR rats that exhibit hypertension but minimal renal injury (SHR-B2 strain) had significantly less renal lymphatic vessel numbers compared to WKY controls (25 ± 2 vs. 28 ± 1; p<0.05) and normal levels of macrophages. The signals for lymphangiogenesis, VEGF-C and its receptor VEGF-R3, were both increased significantly at the protein level in the kidneys of SHR-A3 rats at 18 weeks but not different in the kidneys of SHR-B2 rats compared to WKY controls. To test whether the increased lymphangiogensis is due to hypertension and/or renal inflammation and injury, we obtained kidneys from Fischer 344 rats that exhibit normal blood pressure but develop renal inflammation and injury as they age. Compared to kidneys from control 4-month old Fischer rats, kidneys from 20-month and 24-month old Fischer rats had significantly increased numbers of lymphatic vessels (32 ± 3 vs. 74 ± 1 vs. 110 ± 6, respectively; p<0.05) and this was also associated with increased macrophage infiltration. Protein levels of VEGF-C and VEGF-R3 were increased significantly in 20-month old Fischer rats compared to 4-month old controls. These data together demonstrate that renal immune cell infiltration, inflammation, and injury increases lymphangiogenesis.