bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract P644: Mineralocorticoid receptor activation may contribute to supine hypertension in patients with primary autonomic failure
Authors: Arnold Amy C , Okamoto Luis E , Gamboa Alfredo , Shibao Cyndya A , Black Bonnie K , Raj Satish R , Robertson David , Biaggioni Italo
Published: July 2021

Primary autonomic failure is characterized by disabling orthostatic hypotension; but at least half of these patients have paradoxical supine hypertension. Renin-angiotensin (Ang) mechanisms were not initially thought to contribute to this hypertension as autonomic failure patients often have undetectable plasma renin activity. Despite suppressed renin, plasma aldosterone levels are normal and we recently showed that plasma Ang II is elevated and acts at AT
1
receptors to contribute to supine hypertension in these patients. Since aldosterone and Ang II also bind mineralocorticoid receptors (MR) to elevate blood pressure, we tested the hypothesis that MR activation plays a role in the hypertension of autonomic failure. To test this, we determined the acute effects of the MR antagonist eplerenone (50 mg, PO) versus placebo on supine blood pressure in a randomized, double blind, crossover study. Medications were given at 8:00 PM and blood pressure was recorded q2 hours for 12 hours. Seven autonomic failure patients with supine hypertension completed this study (5 pure autonomic failure, 1 Parkinson’s, 1 multiple system atrophy; 5 male; 68±2 years of age). Eplerenone maximally reduced supine systolic blood pressure by 34±6 mmHg at 8 hours after administration (vs. 6±12 mmHg for placebo, p=0.047), with no effect on volume measures (12-hour urine volume: 927±205 placebo vs. 1247±151 ml eplerenone, p=0.438; nocturnal weight loss: -1.1±0.2 placebo vs. -1.2±0.2 kg eplerenone, p=0.813). These findings suggest that inappropriate MR activation may contribute to hypertension in autonomic failure, and provide rationale for use of eplerenone in treatment of these patients. While still under investigation, the lack of effect on volume measures combined with the rapid time course for blood pressure lowering following eplerenone may suggest extra-renal mechanisms are involved in MR antagonism in autonomic failure.
Abstract 055: Systemic Effect of Renal 11β-HSD2 Deficiency on Blood Pressure Regulation
Authors: Ueda Kohei , Nishimoto Mitsuhiro , Hirohama Daigoro , Ayuzawa Nobuhiro , Kawarazaki Wakako , Watanabe Atsushi , Shimosawa Tatsuo , Loffing Johannes , Zhang Ming-Zhi , Marumo Takeshi , Fujita Toshiro
Published: July 2021

Background:
Renal mechanism of 11β-HSD2 deficiency for developing hypertension is to be evaluated because vascular mechanism associated with sympathetic nervous activity was prevailing in global
Hsd11b2
knockout (KO) mice although brain-specific KO mice needed high salt intake to develop hypertension. We have demonstrated the importance of renal 11β-HSD2 deficiency on developing hypertension by using kidney-specific
Hsd11b2
knockout (

Hsd11b2
Ksp-/-

; KS-KO) mice (
Hypertension
, in press) and have continued the analysis of the systemic effect of renal 11β-HSD2 deficiency.

Method:
Blood pressure (BP) and heart rate (HR) was measured by using 24h telemetry. Amiloride (25 mg/L) and hydrochlorothiazide (HCTZ, 300 mg/L) were administered through drinking water. Pellet containing MR antagonist spironolactone (MRA; 50 mg/KgBW/day) was administered subcutaneously. Corticosterone concentration was determined by ELISA. Data are presented as mean±SE.

Result:
Systolic and diastolic BPs of KS-KO mice were significantly higher, although the HR was lower, than those of WT mice: SBP, 142.4±1.0 vs 122.4±0.8 mmHg; DBP, 103.9±0.8 vs 94.4±0.8 mmHg; HR, 492.5±2.7 vs 555.4±2.4 (n=7). Mean BP was decreased to the level of WT mice by reducing dietary sodium content from 0.3 % to 0.01 %. Plasma [K
+
] was significantly lower in KS-KO mice: 2.9±0.2 vs 4.2±0.2 mEq/L (n=5). Renal membrane expressions of NCC, T53-phosphorylated NCC (pNCC), cleaved ENaCα and full-length ENaCα were upregulated in KS-KO mice. Correction of plasma [K
+
] of KS-KO mice by using high KCl diet or amiloride downregulated the renal membrane expression of pNCC and decreased the MBP to the level of WT mice as well as chronic HCTZ-treated KS-KO mice. Subcutaneous administration of MRA decreased MBP of KS-KO mice and the renal membrane expressions of pNCC and cleaved ENaCα. Diurnal variation of plasma corticosterone concentration was diminished in KS-KO mice and the urinary excretion of corticosterone was higher compared to that in WT mice.

Conclusion:
Renal 11β-HSD2 deficiency is sufficient in developing hypertension via MR activation induced by excessive corticosterone, the systemic effect of which are also suggested.
Abstract 061: Loss of Lymphocyte Adaptor Protein LNK Promotes Acute Aortic Dissection
Authors: Laroumanie Fanny C , Saleh Mohamed A , Dale Bethany L , Gavulic Kyle A , McMaster William G , Madhur Meena S
Published: July 2021

Background:
Acute aortic dissection (AD) is a life-threatening vascular disease associated with an inflammatory response. A polymorphism in the gene
SH2B3
that encodes LNK has been associated with several cardiovascular and autoimmune diseases in humans. LNK is an adaptor protein expressed in hematopoietic and endothelial cells that serves as a brake to cellular proliferation and cytokine production. We hypothesize that loss of LNK promotes AD through an exacerbation of the acute immune response.

Methods:
Angiotensin II (Ang II) was infused for 3 or 14 days into wild type (WT) and LNK
-/-
mice. Kaplan-Meier survival curves were generated. After 3 days, the aortic remodeling was accessed by standard histological staining methods and microscopy. The aortic inflammation was characterized by flow cytometry and immunohistochemistry.

Results:
Ang II infusion induces a rapid and drastic mortality in LNK
-/-
mice compared to WT mice (66% vs 8%,
P
<0.001). Necropsies revealed that deaths are due to the development of AD or rupture, localized primarily in the abdominal aorta. Interestingly, during the phase that precedes AD development (day 3 of Ang II infusion), the aortas of LNK
-/-
mice show significant remodeling with more elastin fragmentation than WT mice (10.5 vs 4.6 breaks,
P
<0.01) and less adventitial collagen deposition (3.3 vs 4.6 x10
4
μm
2
,
P
=0.057). Strikingly, collagen qualitative analysis reveals thinner collagen fibers and several areas of disruption and disorganization in the aorta of LNK
-/-
mice prior to AD development. In parallel, the aorta of LNK
-/-
mice show an increase in the number of neutrophils and macrophages but not in T cells compared to WT mice.

Conclusion:
In our model, LNK seems to play a key role in maintaining the aortic wall integrity. Loss of LNK promotes acute inflammation and matrix degradation, leading to the development of AD. Targeting LNK could be a potential therapeutic strategy for the management of aortic dissection.
Abstract 070: Canonical Wnt Signaling Mediates Enhanced Renal Afferent Arteriolar Reactive Oxygen Species and Contractility in Diabetic Mice
Authors: Lai En Yin , Zhang Suping , Huang Qian , Wang Qiaoling , Zhao Liang , Patzak Andreas , Li Lingli , Wilcox Christopher S
Published: July 2021

Background:
Canonical Wnt signaling is involved in oxidative stress and diabetes but its role in diabetic renal microvascular dysfunction is unclear. We tested the hypothesis that enhanced canonical Wnt signaling in renal afferent arterioles from diabetic mice increases reactive oxygen species (ROS) and contractions to endothelin-1 (ET-1).

Methods:
Diabetic or control C57Bl/6 mice received vehicle or sulindac (40 mg·kg
-1
·day
-1
) to block canonical Wnt signaling for 4 weeks. ET-1 contractions were measured in diameter changes and H
2
O
2
and O
2
.-
by fluorescence microscopy. Arteriolar protein expression and enzymatic activity were examined by standard methods.

Results:
Compared to control, diabetic mouse afferent arteriole had significantly increased O
2
.-
(+84%) and H
2
O
2
(+91%) and enhanced sensitivity to ET-1 at 10
-8
mol·l
-1
(-72±4% versus -43±4%, P<0.05) accompanied by significantly (P<0.005) reduced protein expressions and activities for catalase and superoxide dismutase 2 (SOD2). Incubation of afferent arterioles from normal or diabetic mice with PEG-SOD reduced responses to ET-1 whereas incubation with PEG-catalase reduced sensitivity to ET-1 selectively in arterioles from diabetic mice. The arteriolar protein expressions for canonical Wnt signaling indicated overactivation of this pathway in diabetic mice (2.6-fold increase in p-GSK-3β/GSK-3β and 3.3-fold decrease in p-β-catenin/β-catenin). Sulindac given to diabetic mice normalized the canonical Wnt signaling protein and arteriolar O
2
.-
, H
2
O
2
and ET-1 contractions while doubling (P<0.05) microvascular catalase and SOD2.

Conclusions:
Increased ROS, notably H
2
O
2
, mediated by canonical Wnt signaling contributes to enhanced afferent arteriolar sensitivity to ET-1 in diabetes. Thus, antioxidant pharmacological strategies targeting canonical Wnt signaling may improve vascular function in diabetic nephropathy.
Abstract 079: Dipeptidyl Peptidase IV (DPP4) Inhibition Enhances the Vasoconstrictor Response to Neuropeptide Y During Angiotensin-converting Enzyme Inhibition
Authors: Hubers Scott , Wilson Jessica , Brown Nancy
Published: July 2021

Background:
Dipeptidyl peptidase IV (DPP4) inhibitors are antidiabetic medications that may increase risk of heart failure. Neuropeptide Y (NPY), a substrate of DPP4, is co-released with norepinephrine (NE) and causes vasoconstriction via the Y1 receptor. We tested the hypothesis that DPP4 inhibition would potentiate the effect of exogenous NPY on forearm blood flow (FBF).

Methods:
Seven healthy non-smokers participated in a randomized, double-blinded, placebo-controlled crossover study. Subjects underwent two study days and received sitagliptin 100 mg daily or placebo for seven days before each study day. On each study day, NPY was infused at 0.1, 0.3, 1.0, and 3.0 nmol/min through the brachial artery and FBF was measured using plethysmography. Following 90-minute washout, subjects received intra-arterial enalaprilat and NPY infusion was repeated. Venous and arterial samples were obtained for NE and NPY.

Results:
FBF decreased to a similar extent with increasing doses of NPY during sitagliptin and placebo (Figure 1). During enalaprilat, sitagliptin potentiated the vasoconstrictor effect of NPY compared to placebo (FBF 1.32 vs 1.94 mL/min/100mL at 0.3 nmol/min NPY during sitagliptin and placebo, respectively, P=0.039, Figure 1). NE decreased with ACE inhibition during NPY infusion, but this effect was not altered by DPP4 inhibition (Change in NE -47 vs -66 pg/mL with sitagliptin and placebo, respectively, P=0.92).

Conclusion:
DPP4 inhibition potentiates the vasoconstrictor effect of NPY in the forearm vasculature in the setting of ACE inhibition. These findings have implications for the cardiovascular effects of DPP4 inhibitors in patients receiving ACE inhibitors.
Abstract 085: CHIP: E3 Ubiquitin Ligase Mediates Proteasomal Degradation of Neuronal Nitric Oxide Synthase in the Paraventricular Nucleus of Rats with Heart Failure
Authors: Sharma Neeru M , Katsurada Kenichi , Liu Xuefei , Patel Kaushik P
Published: July 2021

The exaggerated sympathetic drive is a characteristic of heart failure (HF) due to reduced neuronal nitric oxide synthase (nNOS) within the paraventricular nucleus (PVN). Previously we have shown that there were increased accumulation of nNOS-ubiquitin (nNOS-Ub) conjugates in the PVN of rats with HF (1.0±0.05 Sham vs. 1.29±0.06 HF) due to the increased levels of PIN (a protein inhibitor of nNOS, known to dissociate nNOS dimers into monomers) (0.76±0.10 Sham vs. 1.12±0.09 HF) and decreased levels of tetrahydrobiopterin (BH4): a cofactor required for stabilization of nNOS dimers (0.62±0.02 Sham vs. 0.44±0.03 HF). We also showed that there is blunted nitric oxide-mediated inhibition of sympathetic tone via the PVN in HF. Here we examined whether CHIP(C-terminus of Hsp70 -interacting protein), a chaperone-dependent E3 ubiquitin-protein isopeptide ligase known to ubiquitylate Hsp90-chaperoned proteins could act as an ubiquitin ligase for nNOS in the PVN. Immunofluorescence studies revealed colocalization of nNOS and CHIP in the PVN indicating their possible interaction. CHIP expression was increased by 50% in the PVN of rats with HF(0.96±0.08 Sham vs.1.44±0.10* HF). It is shown that Hsp90 protects nNOS from ubiquitination while Hsp70 promotes the ubiquitination and degradation. We observed significant upregulation of Hsp70 (0.49±0.03 Sham vs. 0.65±0.02* HF) with a trend toward the decrease in Hsp90 expression (0.90±0.07 Sham vs. 0.71±0.06 HF). The opposing effects of the two chaperones could account for the increased CHIP-mediated ubiquitination and degradation of dysfunctional nNOS monomers in the PVN of rats with HF. Furthermore, neuronal NG108-15 cell line transfected with the pCMV3-CHIP-GFP spark (CHIP overexpression plasmid) showed approximately 74% increase in CHIP with concomitant 49% decrease in nNOS expression.
In vitro
ubiquitination assay in NG108 cells transfected with pCMV-(HA-Ub)
8
and pCMV3-CHIP-GFP spark plasmid reveal increased HA-Ub-nNOS conjugates (1.13 ± 0.09 Scramble vs. 1.65 ± 0.12* CHIP plasmid). Taken together, our results identify CHIP as an E3 ligase for ubiquitination of dysfunctional nNOS and CHIP expression is augmented during HF leading to increased proteasomal degradation of nNOS in the PVN.
Abstract 101: Tissue-specific Deletion of Collectrin in the Proximal Tubular Epithelium Increases Arterial Pressure and Augments Salt-sensitivity
Authors: Cechova Sylvia , Chu Pei-Lun , Gigliotti Joseph C , Chan Fan , Le Thu H
Published: July 2021

Background:
Collectrin (
Tmem27
) is a key regulator of blood pressure (BP) and modulator of the bioavailability of nitric oxide (NO) and superoxide. It is highly expressed in the kidney in the proximal tubule (PT), collecting duct, and throughout the vascular endothelium. We reported that collectrin plays a critical role as a chaperone for the reabsorption of all amino acids (AAs) in the PT, and for the uptake of the cationic AA L-arginine (L-Arg) in endothelial cells. Global collectrin knockout (

Tmem27
Y/-

) mice display baseline hypertension (HTN), augmented salt-sensitive hypertension (SSH), and decreased renal blood flow.

Objective and Methods:
To determine the PT-specific effect of collectrin on BP homeostasis and salt sensitivity, we used the Cre
-loxP
approach and PEPCK-Cre to generate a mouse line lacking collectrin specifically in the PT-- PEPCK-Cre
+
Tmem27

Y/Flox

mice. PEPCK-Cre
-
Tmem27

Y/Flox

mice were used as control. Radiotelemetry was used to measure BP for 2 weeks at baseline and 2 weeks on high salt diet (HSD). Renal blood flow at baseline and on HSD was measured using contrast enhanced ultrasound in the same mice.

Results:
Successful deletion of collectrin in the PT was confirmed by assessing mRNA levels using real-time RT-PCR, immunohistochemistry staining of renal tissues using anti-collectrin antibody, and quantitation of protein from kidney cortex by Western analysis. Compared to control PEPCK-Cre
-
Tmem27

Y/Flox

mice (n=6), PEPCK-Cre
+
Tmem27

Y/Flox

mice (n=6) displayed significantly higher systolic BP (SBP) at baseline (120.0 ± 2.5 vs 131.6 ± 2.9 mm Hg; p = 0.014) and after HSD (135.3 ± 2.6 vs 151.5 ± 5.2 mm Hg; p = 0.019). Renal blood flow was not different between groups, at baseline nor after HSD.

Conclusion:
Collectrin in the PT plays an important role in blood pressure homeostasis and response to sodium intake, independent of renal blood flow. Increasing proximal tubular collectrin activity may be a novel therapeutic strategy for the treatment of hypertension and salt-sensitivity.
Abstract 104: Cerebrovascular and Cognitive Dysfunction in DOCA-Salt Hypertension is Mediated by Perivascular Macrophages
Authors: Santisteban Monica M , Faraco Giuseppe , Racchumi Gianfranco , Anrather Josef , Iadecola Costantino
Published: July 2021

Hypertension (HTN) and high-salt diets are important risk factors for stroke and dementia. DOCA-salt is a recognized model of HTN driven by sodium retention and brain renin-angiotensin system (RAS) activation. However, it is unknown whether essential mechanisms regulating the cerebral circulation are altered in DOCA-salt mice, and, if so, whether these alterations are associated with cognitive impairment. To this end, C57BL/6 mice were implanted with 50mg DOCA pellets SQ and received 0.9% NaCl drinking water for 3 weeks. Cerebral blood flow (CBF) was measured in the somatosensory cortex by laser-Doppler flowmetry through a cranial window. DOCA-salt increased systolic blood pressure (BP; 148±3 vs 112±3 mmHg in controls; p<0.01), and attenuated the CBF increase induced by whisker stimulation (WS; 16.0±1.1 vs 22.4±0.6 %; p<0.01) or by cortical application of acetylcholine (ACh; 13.5±0.9 vs 22.8±1.1 %; p<0.01), without affecting the response to the smooth muscle relaxant adenosine. Cerebrovascular dysfunction was associated with cognitive impairment as assessed by Novel Object Recognition and Barnes Maze tasks (p<0.01). Perivascular macrophages (PVM) express AT1R and Nox2, and, as such, may be a key source of radicals mediating the cerebrovascular effects of brain RAS overactivity. To test this hypothesis, brain PVM were depleted by icv administration of clodronate (CLO) liposomes. BP was not affected by CLO in either control or DOCA mice (p>0.05). PVM depletion improved novel object exploration (p<0.01) and time spent in the target quadrant of Barnes Maze (p<0.05), while also restoring the CBF responses to both WS (DOCA-CLO 19.6±0.9%; p<0.05) and ACh (DOCA-CLO 20.0±1.7%; p<0.05). Next, we tested whether reactive oxygen species (ROS) are involved in the cerebrovascular dysfunction. We observed a 45% upregulation in
gp91
mRNA in cerebral vessels from DOCA mice, which was prevented by PVM depletion (p<0.05). Application of the ROS scavenger MnTBAP rescued CBF responses to both WS (20.3±0.9%; p<0.05) and ACh (19.0±0.8%; p<0.05) in DOCA-salt HTN. We conclude that PVM play a previously unrecognized role in the cerebrovascular and cognitive dysfunction of DOCA-salt HTN and may represent a new therapeutic target to alleviate the neurocognitive effects of HTN.
Abstract 105: Therapeutic Suppression of Mtorc2 Signaling Reduces Salt-induced Hypertension and Kidney Injury in SS Rats
Authors: Kumar Vikash , Evans Loiuse , Wollner Clayton , Kurth Theresa , Cowley Allen W
Published: July 2021

The present study explored the protective effect of mTORC2 inhibition in salt-induced hypertension and kidney injury. We have previously reported that enhanced blood pressure salt-sensitivity with renal chronic interstitial infusion of H
2
O
2
(347 nmol/kg/min) in normotensive Sprague Dawley (SD) rats. In the present study,
in vivo
experiment was performed in which H
2
O
2
(347 nmol/Kg/min) was chronically infused for 3 days into renal interstitium of unilaterally nephrectomized SD rats. A significant increase of mTORC2 activity (pAKT/AKT) was observed in the renal cortex of SD rats infused with H
2
O
2
compared to saline infused rats. We have recently shown that excess production of H
2
O
2
in the SS rat kidneys is the hallmark of salt-induced hypertension. We hypothesized that mTORC2 in the kidney contributes to the development of salt-induced hypertension in SS rats. Rats were treated with PP242 which is an ATP-competitor inhibitor which inhibits the activities of both mTORC1 and mTORC2 whereas rapamycin specifically inhibits mTORC1. PP242 was administrated daily (i.p., 15 mg/Kg/day) for 21 days to SS rats fed a 4.0% NaCl diet. Remarkably, salt-induced hypertension was significantly reduced in SS rats which averaged 119 ± 2 mmHg in PP242 treated rats (n=7) compared to 168 ± 3 mmHg in vehicle treated rats (n=7). Albuminuria was greatly reduced with urine albumin excretion (mg/day) averaging 32.8 ± 3 in PP242 treated rats compared to 256 ± 37 in vehicle treated rats. PP242 treatment notably resulted in reduced infiltration of T lymphocytes in the kidneys of SS rats fed a 4.0% NaCl diet. CD3
+
cells/mm
2
averaging 157.0 ± 40.0 compared to 36.0 ± 11.0 in the renal cortex and 218.0 ± 24.0 compared to 24.0 ± 9.0 in the outer medulla in PP242 versus vehicle treated rats. These data show that mTORC2 is required for the intiation of salt-induced hypertension and therapeutic suppression of this pathway virtually abolished salt-sensitivity blood pressure and kidney injury.
Abstract 112: Myeloid Mineralocorticoid Receptor Controls Inflammatory and Fibrotic Responses After Renal Ischemic Injury via Macrophage Interleukin-4 Receptor
Authors: Barrera-Chimal Jonatan , Lechner Sebastian M , Moghrabi Soumaya E , Kolkhof Peter , Jaisser Frédéric
Published: July 2021

Introduction:
Patients who survive an episode of acute kidney injury (AKI) are at high risk of de novo chronic kidney disease (CKD) development. Pharmacological mineralocorticoid receptor (MR) antagonism is useful to prevent CKD after a single episode of ischemic AKI in the rat.

Objective:
Test the involvement of myeloid MR in the development of kidney fibrosis after an ischemic AKI episode.

Methods:
We included 18 male C57/B6 mice that were divided in: sham, renal ischemia for 22.5 min and IR plus treatment with the non-steroidal MR antagonist finerenone (10 mg/kg) at -48, -24 and -1 h before IR. MR inactivation in myeloid cells (MR
MyKO
) was achieved by crossing mice with the MR alleles flanked by loxP sites (MR
f/f
) with mice expressing the Cre recombinase under the LysM promoter activity. In MR
f/f
and MR
MyKO
mice we induced renal IR of 22.5 min or sham surgery. The mice were followed-up during 4 weeks to test for AKI to CKD transition. In another set of mice, the macrophages were sorted from kidneys after 24 h of reperfusion and flow cytometry characterization or mRNA extraction was performed. Thyoglycolate elicited peritoneal macrophages were used for
in vitro
studies.

Results:
The progression of AKI to CKD after 4 weeks of renal ischemia in the untreated C57/B6 and MR
f/f
mice was characterized by a 50% increase in plasma creatinine, a 2-fold increase in the mRNA levels of TGF-β and fibronectin as well as by severe tubule-interstitial fibrosis. The mice that received finerenone or MR
MyKO
mice were protected against these alterations. Increased expression of M2-anti-inflamatory markers in kidney-isolated macrophages from finerenone-treated or MR
MyKO
mice was observed. The inflammatory population of Ly6C
high
macrophages was reduced by 50%. In peritoneal macrophages in culture, MR inhibition promoted increased IL-4 receptor expression and activation, facilitating macrophage polarization to an M2 phenotype.

Conclusion:
MR antagonism or myeloid MR deficiency facilitates macrophage polarization to a M2, anti-inflammatory phenotype after kidney IR, preventing maladaptive repair and chronic kidney fibrosis and dysfunction. MR inhibition acts through the modulation of IL-4 receptor signaling to facilitate macrophage phenotype switching.
Abstract 113: Beneficial or Harmful? The Role of Intensive Anti-hypertensive Treatment in the Development of Chronic Kidney Disease
Authors: Wang Ling , Wang Donna
Published: July 2021

Hypertension is the leading cause of end-stage renal disease, and one of the goals of anti-hypertensive treatment is to protect the kidney. However, it is unknown how low of blood pressure as the treatment target should be so that anti-hypertensive therapy would not bring harm to patients especially for those already suffer from chronic kidney disease (CKD). Thus, we used the data set from The Systolic Blood Pressure Intervention Trial (SPRINT) to study the effect of lowering systolic blood pressure on renal disease development. The SPRINT data randomly assigned patients with a systolic blood pressure (SBP) of 130 mm Hg or higher to a SBP treatment target of less than 120 mm Hg (intensive treatment, n=4678) or a treatment target of less than 140 mm Hg (standard treatment, n=4683). We examined the effect of intensive treatment on six renal outcomes: 1) CKD composite, 2)50 percent reduction in eGFR, 3) dialysis 4) albuminuria, 5) 30 percent reduction in eGFR for patients with CKD at baseline (n=2646) and 6) albuminuria for patients without CKD at baseline (n=6715). Generalized Estimating Equation is used to account the correlation of blood pressure levels over time. At the end of year 1, the mean SBP was 121.4± 0.21 mm Hg in the intensive treatment group and 136.2± 0.21 mm Hg in the standard treatment group. The patients in intensive group were found to have a higher chance of 30% reduction of eGFR (OR=3.684, 95% CI= 2.51-5.40) than in standard treatment group. There was no difference between intensive and standard treatment groups for other 5 outcomes. In addition, 1 mm Hg elevation in SBP in patients with CDK at baseline significantly increased the chance of CKD composite (OR=1.03, 95% CI=1.01-1.04), the chance of 50 percent reduction in eGFR (OR=1.02, 95% CI=1.01-1.05), and chance of 30 percent reduction in eGFR (OR=1.02, 95% CI=1.01-1.02). Thus, SBP significantly correlated with renal outcomes in CKD patients. Our data show that five renal outcomes examined using SPRINT data set are not improved by intensive management of SBP in CKD patients, rather, patients received intensive management have a higher risk of eGFR reduction by 30%, which could be detrimental. Our study indicated that intensive SBP management should not be recommended to CKD patients.
Abstract 135: PVN Galphai2 Protein Over Expression Attenuates Dahl Salt Sensitive Hypertension and GNAI2 Polymorphic Variance Represents a Clinical Biomarker of the Salt Sensitivity of Blood Pressure
Authors: Wainford Richard D , Zhang Xiaoling
Published: July 2021

Aim:
We hypothesize that a) in the DSS rat, which in contrast to the normotensive DSR rat fails to up regulate PVN Gαi
2
proteins on a high salt intake (Wainford et al, Hypertension 2015), PVN specific Gαi
2
protein over expression will attenuate the development of salt sensitive hypertension (HTN) and, b) SNPs in the GNAI2 gene, which are associated with HTN in Japanese and Italian populations, represent a biomarker of the salt-sensitivity of BP.

Methods:
Male DSS rats instrumented with a radiotelemetry probe (DSI; PA-C40) received a bilateral PVN shuttle or Gαi
2
expressing lenti-viral vector microinjection (2x10
9
infectious units per ml/60nl/side) and were maintained on a 7-day normal 0.4% (NS) intake for baseline BP prior to 21 day high 4% NaCl (HS) intake. On day 21 rats were sacrificed to assess PVN Gαi
2
protein expression (immunoblotting), plasma NE content (ELISA) or underwent a 5% volume expansion (VE) prior to cardiac perfusion and c-fos IHC to assess PVN neuron activation (N=4/gp/study). SNPs in the GNAI2 gene were examined in GenSalt (N=968) for associations with the salt sensitivity of BP (increase in systolic BP of 5mmHg or greater during HS intake, N=326 are salt sensitive: 172 Female, 154 male).

Results:
Bilateral lenti-virus increased PVN Gαi
2
protein levels 3 fold (p<0.05) on both NS and HS intake,
attenuated
HTN (Day 21 HS MAP [mmHg] shuttle 160±3 vs. Gαi
2

144±4, P<0.05),
abolished
Na
+
evoked global and renal sympathoexcitation (Day 21 HS plasma NE [nmol/L] shuttle 82±5 vs. Gαi
2
50±4, P<0.05). PVN Gαi
2
over expression increased VE-evoked natriuresis (peak natriuresis (UNaV; [μeq/min] shuttle NS 43±3, shuttle HS 16±3, Gαi
2
HS 31±3, P<0.05) and PVN sympathoinhibitory parvocellular neuron activation (medial parvocellular c-Fos positive cells shuttle NS 67±8, shuttle HS 29±3, Gαi
2
vector HS 48±6, P<0.05). 3 GNAI2 SNPs are present in GenSalt - rs9852677 and rs2282751 did not associate with the salt sensitivity of BP. SNP rs10510755 positively correlated with the salt sensitivity of BP (MAF:6%, Z-score: 1.94, p<0.05) independently of sex.

Conclusion:
These data suggest a critical role of PVN Gαi
2

proteins in countering the pathophysiology of salt sensitive HTN and that GNAI2 polymorphic variance represents a biomarker of the individual salt sensitivity of BP.
Abstract 136: Improvement of BP Control (and Simultaneous Reduction of Drug Consumption) in Treated Hypertensive Patients Compliant to a Minimal Dietary Sodium Restriction
Authors: Musso Natale , Carloni Beatrice , Chiusano Maria C
Published: July 2021

The official guidelines for hypertension recommend a low sodium diet as a basic approach. Sodium should be restricted to 100 mEq a day. Such tight diets are seldom paralleled by an adequate compliance.

Methods:
we studied the effects of a reasonable and realistic low sodium diet on the BP values in 196 pts. (157 pts. completed the study). The patients had to avoid ice creams, cheese and salt-preserved (cured) meat. They were switched from regular to salt-free bread. Otherwise they were free to follow their usual mediterranean diet. Urinary output of Na and K, weight, number of antihypertensive drugs taken, and office BP/HR were measured before diet started (Value 1 in the table) and after 9±1 weeks on diet (Value 2 in the table) (all values are mean ± SD). The BP was recorded with the patient alone, in a seated position, with automatic repeated measures, by means of Omron 907 HEM monitors. At the end of the study 88 patients showed a reduction in the urinary sodium (compliant pts.), whereas 69 did not (noncompliant pts.).

In those compliant to the low salt approach (88 out of 157 pts. i.e. 56%), a substantial improvement was attained in terms of BP control and body weight with a simultaneous significant reduction in the daily drug requirement. The responders increased from 47 to 69 (i.e. from 53% to 78%). In the noncompliant group the BP values, the responder rate (35 to 32, i.e. 50% to 46%), the weight, and the drug consumption were stable.

Conclusion:
even a limited dietary salt reduction may be beneficial in terms of BP control, with a decrease of the BP levels and an increase in the responder rate, paralleled by a reduction of the drug requirement.

Take home message: even a bit of diet is better than no diet.
Abstract 139: Loss of Thymosin b4 Exacerbates Renal and Cardiac Damage in Angiotensin-II Hypertension
Authors: Kumar Nitin , Liao Tang-Dong , Romero Cesar , Maheshwari Mani , Peterson Ed , Carretero Oscar
Published: July 2021

Angiotensin-II (Ang-II)-induced hypertension is associated with tissue damage and fibrosis in the kidney and heart. Thymosin β4 (Tβ4) regulates cell morphology, inflammation and fibrosis in several organs and administration of exogenous Tβ4 is protective in diabetic nephropathy and unilateral ureteral obstruction model. However, role of endogenous Tβ4 in hypertension-induced organ damage is unknown. We
hypothesize
that, loss of Tβ4 accelerates renal and cardiac fibrosis and damage in Ang-II hypertension. To test our hypothesis, Tβ4 knockout (Tβ4
-/-
) and wild-type (Tβ4
+/+
) C57BL/6 mice (n=6-10) were infused continuously for six-weeks with either Ang-II (980 ng/kg/min) or vehicle via osmotic minipumps. All the results are presented in table 1. In Ang-II infusion, systolic blood-pressure were not different between both strains (Table 1). Interestingly, urinary albuminuria was significantly higher in Tβ4
-/-
mice compared to Tβ4
+/+
mice by Ang-II. High expression of Tβ4 is found in the glomeruli along with high expression of Nephrin, an important protein in the filtration barrier of the kidney. In Ang-II infusion, nephrin protein expression was greatly reduced in mice deficient of Tβ4, suggesting that loss of nephrin is one of the mechanism for elevated urinary albumin in Tβ4
-/-
mice. Additionally, renal fibrosis was higher in Tβ4
-/-
mice. We also studied cardiac damage and observed that in Ang-II infusion, cardiac hypertrophy and cardiac fibrosis were much higher in Tβ4
-/-
mice. These data indicate that loss of endogenous Tβ4 caused significant tissue damage in the kidney and heart in Ang-II hypertension, suggesting renal and cardiac protective role of this peptide.
Abstract 142: Angiotensin II Type 1 Receptor Autoantibody Inhibition Improves Blood Pressure and Markers of Neurological Damage and Oxidative Stress in Brains of Placental Ischemic Rats During Pregnancy
Authors: Cunningham Mark W , Vaka Venkata Ramana , Amaral Lorena , Fan Fan , Ibrahim Tarek , Roman Richard R , LaMarca Babbette
Published: July 2021

Preeclampsia (PE), hypertension in response to placental ischemia, is associated with angiotensin II type 1 receptor agonistic autoantibodies (AT1-AA), oxidative stress, and neurological complications, such as headaches, blurred vision, and seizures which could lead to stroke and death. We hypothesize that AT1AAs play a role in the cerebral pathology of PE. The objective of this study was to determine if administration of a specific peptide sequence to inhibit the AT1-AA from binding to the AT1 receptor, will improve blood pressure (MAP) and cerebral oxidative stress in the reduced uterine perfusion pressure (RUPP) rat model of PE. Pregnant Sprague Dawley rats were divided into 2 groups: RUPP (n=5) and RUPP+AT1-AA inhibitory peptide (7AA) (n=3). RUPP surgery was performed on gestational day (GD) 14 and the 7AA was administered (2ug/μl saline) via mini-osmotic pumps. On GD 19, MAP was determined and brains collected. Western blots were stained for Glial Fibrillary Acidic Protein (GFAP), endothelial NO synthase (eNOS), phosphorylated eNOS and NADPH oxidase activity was determined using chemilumenescence. MAP was decreased in RUPP+7AA vs. RUPP (95±2 vs. 130±6 mmHg). Brain/body weight ratio, which is indicative of edema, was reduced in RUPP+7AA (5.8±0.25 vs. 6.5±0.25 grams) vs. RUPP. NADPH oxidase activity was lower in RUPP+7AA (33275±3122 vs. 57408±10508 RLU/min/mg protein). Phosphorylated eNOS was 2 fold higher in the RUPP+7AA vs. RUPP (0.4±0.1 vs. 0.2±0.04 AU) and the phosphorylated eNOS/eNOS ratio was elevated (0.4±0.12 vs. 0.2±0.04 AU). GFAP a marker for activated astrocytes that increases during neurologic injury and serves as a compensatory mechanism for brain injury recovery was elevated in RUPP+7AA vs. RUPP (3.2±1.3 vs. 0.5±0.2 AU). Administration of AT1-AA inhibitory peptide to RUPP rats decreased blood pressure and improved markers of NO bioavailability, injury (GFAP), and cerebral swelling. In conclusion, our preliminary data suggests that AT1-AA inhibition could be a potential therapy to improve peripheral and neurological complications during PE.
Research Supported by T32HL105324 (Cunningham), RO1HD067541-06 (LaMarca), DK-104184 (Roman), 050049 (Fan), P20-GM-104357 (cores B and C-Roman; Pilot-Fan) and AHA 16GRNT31200036 (Fan).
Abstract P102: Liver Acquitted, Fat Indicted: Hepatic Chemerin Knockdown Does Not Reduce Blood Pressure While Whole-body Knockdown Does
Authors: Ferland David J , Seitz Bridget , Darios Emma S , Thompson Janice M , Yeh Steve T , Mullick Adam E , Watts Stephanie W
Published: July 2021

Chemerin is an inflammatory adipokine positively associated with hypertension and obesity with the majority of chemerin thought to derive from the liver and adipose tissue. The contributions of liver-derived chemerin to plasma chemerin levels and blood pressure regulation are unknown. We compared whole-body vs liver chemerin inhibition using antisense oligonucleotides (ASO) with liver-restricted activity (GalNAc) or liver and fat activity (Gen 2.5). We hypothesized that in normotensive male SD rats, circulating chemerin is predominately liver-derived and regulates blood pressure. A scrambled ASO control and phosphate-buffered saline (PBS) were used as controls and radiotelemetry was used to monitor blood pressure. Baseline mean arterial blood pressure (MAP) was recorded for one week, beginning 5 days after surgery to establish a baseline. ASOs were given weekly by subcutaneous injections for four weeks. Two days after the final injection, animals were sacrificed for tissue RT-PCR and plasma chemerin measurements using Western analysis. Gen 2.5 chemerin ASO treatments (compared to PBS control) reduced chemerin mRNA in liver, retroperitoneal fat, and mesenteric perivascular adipose tissue (mPVAT) by 99.5% ± 0.1%, 95.2% ± 0.3%, and 69% ± 2%, respectively, and plasma chemerin was reduced to undetectable levels. GalNAc chemerin ASO treatments (compared to PBS control) reduced chemerin mRNA in liver by 97.9% but had no effect on chemerin expression in retroperitoneal fat and mPVAT; plasma chemerin was reduced by 90% ± 5%. Gen 2.5 chemerin ASO treatment reduced MAP, which reached a nadir of 7 ± 2.1 mmHg 48 – 72 hours after each dose compared to the scrambled ASO controls. By contrast, MAP was unchanged in animals receiving the GalNAc chemerin ASO. Thus, although most circulating chemerin is liver-derived, plasma chemerin does not play a role in blood pressure regulation. This study suggests that while chemerin is still generally associated with increased blood pressure, circulating chemerin levels cannot directly predict this effect. In addition, local effects of chemerin from fat may explain this discrepancy and support chemerin’s association with both hypertension and obesity.
Abstract P117: Hydralazine Prevents the Rupture of Experimental Cerebral Aneurysms in Hypertensive Rats
Authors: Miyamoto Takeshi , Kitazato Keiko T , Tada Yoshiteru , Shimada Kenji , Yagi Kenji , Korai Masaaki , Maekawa Hidetsugu , Yamaguchi Tadashi , Kanematsu Yasuhisa , Satomi Junichiro , Hashimoto Tomoki , Nagahiro Shinji
Published: July 2021

Introduction:
Subarachnoid hemorrhage (SAH) is a catastrophic event with high morbidity and a poor prognosis. While epidemiological studies showed that hypertension is a risk factor for aneurysmal rupture, it remains unclear whether lowering the blood pressure (BP) prevents the rupture of aneurysms. Under the hypothesis that lowering the BP prevents the rupture of experimental cerebral aneurysms, we investigated whether hydralazine reduces the incidence and rupture of aneurysms in our rat model.

Methods:
In 10-week-old female Sprague-Dawley rats (n=34) we elicited estrogen deficiency, renal hypertension, and hemodynamic stress. Two weeks later, they were divided into 2 groups; group 1 (n=17) was treated perorally with hydralazine (100 mg/kg/day), group 2 (n=17) was the vehicle control. We recorded their death or abnormal behavior in the course of 90 days and inspected ruptured aneurysms.

Results:
At 2 weeks, both groups manifested an increase in the systolic BP (SBP) (group 1, 205 mmHg; group 2, 207 mmHg). After 30-, 60-, and 90-days, the SBP was lower in group 1 than group 2 (150 vs 210 mmHg, 163 vs 211 mmHg, and 173 vs 210, mmHg, respectively). In the course of 90 days, 9 group 2 (53%) and 8 group 1 rats (47%) developed cerebral aneurysms. The rupture rate was lower in group 1 (3/8, 38%) than group 2 rats (9/9: 100%). While hydralazine did not prevent the development of aneurysms, it prevented their rupture. qRT-PCR performed on day 35 showed the down-regulation of MCP-1 and MMP-9 in the cerebral vascular wall of group 1 rats.

Conclusion:
Our findings suggest that lowering the SBP may prevent the rupture of cerebral aneurysms via the down-regulation of MCP-1 and MMP-9 in the cerebral vascular wall.
Abstract P125: RNA Sequencing of Left-ventricular Tissue Highlights Novel Pathways Impacted by the Loss of miR-146b-5p in a Rat Model of Chronic Kidney Disease
Authors: Paterson Mark R , Liu Pengyuan , Kriegel Alison J
Published: July 2021

Chronic kidney disease (CKD) mediates pathological changes in heart and vascular tissues, yet many molecular mechanisms by which renal injury directly affects cardiovascular pathology (CVD) are unknown. miRNAs are proposed to be powerful regulators of pathophysiological signaling networks; having the potential to radically change the landscape of mRNA transcript expression and protein abundance in the context of health and disease. We have profiled the expression of miRNAs in a rat model of CKD – the 5/6 nephrectomy (5/6Nx) – with the objective to identify miRNAs with strong effects on CKD-mediated CVD. One such miRNA, miR-146b-5p (miR-146b), is highly upregulated in 5/6Nx rats compared to sham-operated controls at 7-weeks post-5/6Nx in both the kidney (5.4 fold increase; p<0.05) and the heart (1.6 fold increase, p<0.05), a time-point at which we see maximal renal and cardiac pathology. To examine the effects of miR-146b upregulation in our model of CKD, we performed RNA sequencing on cardiac tissue at 7 weeks post-5/6Nx in both wild-type (WT) Sprague-Dawley (SD) rats and a unique CRISPR-Cas9-mediated miR-146b knockout strain (miR-146b
-/-
) created on the SD background. Of 3,018 genes differentially regulated by more than 1.5-fold (absolute value) in miR-146b
-/-
vs. WT rats after 5/6Nx, 135 had significantly altered expression (adjusted p-value <0.05). We filtered all differentially expressed genes through Ingenuity Pathway Analysis to identify affected signaling networks in miR-146b
-/-
vs. WT rats that correlate with observed pathological phenotypes. The expression pattern of differentially regulated genes in these networks indicates predicted activation of pathways that contribute to tissue fibrosis and mineralization, reactive oxygen species signaling, and the inflammatory and immune responses. These pathways are enriched with predicted miR-146b target genes and correlate closely with observed phenotypes in miR-146b
-/-
5/6Nx rats; including increased renal fibrosis and mineralization of cardiovascular tissue, and the well-known role of miR-146b in negative feedback regulation of NFκB signaling. These preliminary analyses uncover unique networks of genes regulated by miR-146b that may contribute to CKD-mediated CVD.
Abstract P135: Is Heart Failure With Preserved Ejection Fraction a Gut Disorder?
Authors: Qi YanFei , Goel Ruby , Mandloi Avinash S , Kim Seungbum , Lobaton Gilberto O , Aquino Victor P , Leach Dana D , Handberg Eileen M , Raizada Mohan K , Pepine Carl J
Published: July 2021

Background and Objective:
Heart failure (HF) with preserved ejection fraction (HFpEF) accounts for ~50% of HF hospitalizations, is associated with poor quality of life and, has a mortality rate comparable with many cancers. To date, no therapies have proven effective in slowing disease progression, mainly due to limited understanding of underlying mechanisms. Mounting evidence suggests the intestine and gut microbiota play an important role in chronic inflammation in HF, and also hypertension, a history of which is present in most HFpEF patients. However, information about intestinal involvement in HFpEF is limited. Accordingly, we investigated the hypothesis that disturbed intestinal barrier function contributes to translocation of endotoxin and inflammation activation in HFpEF.

Method and Results:
Left ventricular (LV) function, intestinal permeability, circulating lipopolysaccharide (LPS, an endotoxin and strong inducer of pro-inflammatory cytokines) and high-mobility group box protein (HMGB1, a nuclear protein that triggers inflammation) were examined in a mouse model of HFpEF [4w continuous infusion of subpressor dose of angiotensin II (0.2mg/kg/d)] and 4 patients with HFpEF and 4 reference subjects. Impaired LV diastolic function (increase in LVEDP and Tau, both p<0.05) occurred in the mice with 3-fold increased intestinal permeability, 1.8-fold elevated LPS levels (137±7 vs control 77±6 pg/ml, p<0.005) and 4-fold increased HMGB1 levels (238±42 vs control 56±16 ng/ml, p<0.05). Stimulating human coronary artery endothelial cells with 10μg/ml LPS for 48h resulted in 3-fold elevation of HMGB1 (29±4 vs control 12±3 ng/ml, p<0.05) that was suppressed by butyrate (17±3 ng/ml). Patients with HFpEF had a 2-fold increase in zonulin (31±3 vs 14±2 ng/ml) and 4-fold LPS elevation (152±18 vs 32±3 pg/ml), (both p<0.05 vs reference subjects), confirming gut barrier dysfunction with translocation of endotoxin.

Conclusions:
HFpEF is associated with increased gut permeability that facilitates LPS translocation activating inflammation resulting in endothelial damage. Thus, the gut could be a novel target for therapeutic interventions in patients with HFpEF.
Abstract P147: Differences in the Treatment of Resistant Hypertension in African Americans and European Americans in a Clinical Setting
Authors: Shuey Megan M , Brown Nancy J , Denny Joshua C
Published: July 2021

Using Vanderbilt University Medical Center’s electronic health record (EHR), we tested the hypothesis that we could identify patients with resistant hypertension (RH) and patterns of treatment of RH in a real-world clinical setting.

Patients were identified as having RH if they had blood pressure (BP) >140/90 mmHg despite concurrent treatment with three different classes of antihypertensive medications including a thiazide diuretic or amlodipine or similar dihydropyridine (DHP) calcium channel blocker (CCB) or if they were treated with four antihypertensive medications including the same classes. Secondary causes and chronic kidney disease were excluded. Among 186,015 European American (EA) and 33,576 African American (AA) hypertensive patients, 13,541 (7.3%) and 3,541 (10.5%) had RH, respectively. AA with RH were younger, heavier, more often female, had a higher incidence of type 2 diabetes, and had higher systolic and diastolic BPs than EA with RH. AA with RH were more likely than EA to be treated with vasodilators, DHP CCB, and α2-agonists. EA were more likely to be treated with angiotensin receptor blockers, renin inhibitors, and beta-blockers. Mineralocorticoid receptor (MR) antagonists were used in both EA and AA patients and use was increased in patients treated with
>
four antihypertensive medications compared to patients treated with three (2.6% vs 12.4% in EA, p<0.001; 2.8% vs 12.3% in AA, p<0.001). The number of patients treated with an MR antagonist increased to 36.6% in EA and 40.3% in AA over a mean follow-up period of 7.4 and 8.7 years, respectively.

Our results demonstrate that clinicians use different classes of medication to treat RH in AA and EA.
Abstract P148: Effect of Apparent Treatment Resistant Hypertension on Cardiovascular Disease Events and Modification by Sex
Authors: Nelson Julianne , Liu Longjian
Published: July 2021

Apparent treatment resistant hypertension (ATRH) is an important health concern in the U.S. affecting approximately 9% of all hypertensive adults and growing. However, long-term prognosis of those with ATRH remains to be fully investigated, especially with regard to the presence of differences by sex. Using data from the Antihypertensive and Lipid-Lowering Treatment to Prevent Heart Attack Trial (ALLHAT) we examined the relationship between ATRH and CVD and effect modification by sex. Subjects in ALLHAT were 55 years or older (N male = 13,597 and N female = 11,919) and were randomized to one of four antihypertensive medications for the purposes of the trial. We used Cox Proportional Hazard models and tested for interaction of sex through use of a multiplicative interaction term, stratified analyses, the Aalen Additive Hazards model, and joint effects. Missing blood pressure readings were imputed using multiple imputation as a sensitivity analysis. Due to the design of ALLHAT, ATRH was assessed at the year 2 follow-up visit and subjects were then followed for an additional 6 years (average follow-up 4.7 years) during this time there were 5,030 CVD events. Overall (N=25,516), ATRH was associated with an approximate 30% increased risk of CVD (HR 1.30, 95% CI 1.19 – 1.42) compared to those without ATRH. This risk was greater in women than in men (p
interaction
<.0001). In women, ATRH was associated with an approximate 62% increase in risk of CVD (HR 1.62, 95% CI 1.41 – 1.86) while in men, ATRH was associated with an approximate 13% increase in risk of CVD (HR 1.13, 95% CI 1.01 – 1.27). Sex was also a modifier of the relationship on the additive scale (p-value 0.003). ATRH is associated with an increased risk of developing CVD and women with ATRH were at a greater risk of developing CVD compared to men. These findings provide important insights into the complex relationship between ATRH and cardiovascular health, and suggest that women, in particular, may be a high-risk subgroup. Thus future studies should examine these differences to aid in development of targeted treatment and interventions which would have a significant public health benefit.