bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 356: Torcetrapib and Dalcetrapib, CETP inhibitors, Induce Adipocyte-derived Aldosterone and Reactive Oxygen Species Formation
Authors: Rios Francisco J , Nguyen Dinh Cat Aurelie , Palacios Roberto , Carswell Andrew , Montezano Augusto C , Touyz Rhian M
Published: March 2022

Hyperaldosteronism and hypertension were major side effects observed in clinical trials with Torcetrapib (TORC), a CETP inhibitor that increases HDL. Reasons for this are unclear. Given that CETP inhibitors accumulate in adipose tissue and that adipocytes are a source of aldosterone, we questioned whether CETP inhibitors, TORC and Dalcetrapib (DALC), influence adipocyte aldosterone production. Putative mechanisms underlying this were also explored. Human adipocytes, expressing CETP were studied and compared to mouse adipocytes, which lack CETP gene. Human SW872 and 3T3-L1 adypocites were treated with TORC and DALC (1-30 μM). Reactive Oxygen Species (ROS) production was evaluated by lucigenin-enhanced chemiluminescence. Aldosterone levels were evaluated by ELISA. Gene expression for Mineralocoricoid Receptor (MR), PPARγ, aldosterone synthase (CYP11B2) and glucorticoid synthase (CYP11B1) were evaluated by real time PCR. In human adipocytes, TORC and DALC induced 2- and 1.5-fold increase respectively in ROS production after 5 min (p<0.05). Aldosterone production was observed after 5h stimulation for both drugs (TORC: 72.9 pg/mL; DALC: 78 pg/mL vs cont 49.7 pg/mL, p<0.05). DALC increased mRNA expression of CYP11B1 (40%), CYP11B2 (40%), GR (80%), MR (80%) and PPARγ (90%) (p<0.05). TORC increased the mRNA expression for CYP11B2 (37%) and PPARγ (41%) (p<0.05) and did not change the mRNA for CYP11B1 or MR. In mouse adipocytes, TORC and DALC induced aldosterone production (120 pg/mL and 117 pg/mL vs cont 79 pg/mL, p<0.05) but only DALC induced ROS production, 1.6-fold increase (p<0.05 vs cont). Both drugs induced aldosterone production in adipocytes and exerted different effects regarding adypocyte gene expression. These effects may not be related to CETP inhibition, since they were also observed in mouse cells. Our findings indicate that CETP inhibitors influence adipocyte function and stimulate adipocyte-derived aldosterone production. We also show that off-target activities of TORC and DALC are not necessarily due to CETP inhibition and might be dependent on ROS production. These novel findings have important clinical significance and may explain, in part, the aldosteronism/hypertension reported in CETP clinical trials.
Abstract 36: Adult Hypertension Risk is More than Quadrupled in Obese Children
Authors: Watson Sara E , Hannon Tamara S , Eckert George J , Pratt J Howard , Rosenman Marc , Tu Wanzhu
Published: March 2022

There is a known association between adiposity and hypertension (HTN). However, whether and to what extent body mass index (BMI) predicts adult HTN has not been well examined longitudinally. In this study, we attempt to quantify the effects of childhood overweight and obesity on adult HTN in a prospective cohort.

The original cohort was established in 1986, and consisted of 1117 healthy children (47% male), recruited from schools in Indianapolis, Indiana (mean age 12.1; SD 5.7 years). During the course of follow-up, blood pressure, height, and weight were measured semiannually with a mean of 8.4 visits per child (SD 7.2). BMI was calculated from recorded height and weight; weight status was determined by age and sex-adjusted BMI percentile values (BMI%). Subjects were classified as normal weight, overweight (BMI% ≥ 85% and < 95%), or obese (BMI% ≥ 95%) based on either their mean BMI % or maximum BMI % during childhood. Evaluation based on mean childhood BMI % revealed that 765 (68%) were normal weight, 176 (16%) were overweight, and 176 (16%) were obese. The mean age of the subjects as of 2013 was 33.4 years (SD 3.9). From the cohort, we identified 119 adult cases of HTN through active follow up and medical chart review. In those with normal weight as children, 6% were hypertensive as adults. The rate of HTN was higher for those classified as overweight or obese as children (14% and 26% respectively, p = <0.0001). Children classified as overweight or obese had double and quadruple the risk to have a diagnosis of HTN in adulthood, respectively, as compared to normal weight children [OR = 2.2 (95% CI = 1.3, 3.6), OR = 4.4 (95% CI = 2.8, 6.9) respectively]. Additional measures of adiposity were similarly associated with HTN in adulthood including waist circumference and tricep skinfold. This pattern was also seen when analyzed by maximum BMI%. However, the increased risk of HTN in adulthood for children classified as overweight or obese was not as great [OR = 1.3 (95% CI = 0.7, 2.3), OR = 3.7 = (2.4, 5.7) respectively].

In conclusion, the degree of adiposity in childhood significantly impacts the risk for HTN in adulthood. Childhood obesity is associated with a quadrupled risk of adult HTN. This has significant implications for early prevention of childhood obesity and public health.
Abstract 364: Maternal History of Hypertension and Incident Atrial Fibrillation According to Blood Pressure Category in a General Urban Population: The Suita Study
Authors: Kokubo Yoshihiro , Watanbe Makoto , Kobayashi Takashi , Kamakura Shiro , Kawanishi Katsuyuki , Miyamoto Yoshihiro
Published: March 2022

Objective:
Atrial fibrillation (AF) is the most common tachyarrhythmia and is a strong risk factor for stroke. However, few prospective studies have examined the relation between a parental history of hypertension and the incidence of AF in a general population. We assessed the hypothesis that a parental history of hypertension is associated with an increased risk of incident AF in an urban general population in Japan.

Methods:
A total of 6,918 participants (30~84 years old) initially free of AF were prospectively followed up for incident AF in the Suita Study. Standard 12-lead electrocardiograms were obtained from all subjects in the supine position. Each record was coded independently by 2 well-trained physicians using the Minnesota Code. Participants were diagnosed with AF if AF or atrial flutter was present on electrocardiograms obtained during a biannual routine health examination or if AF was indicated as a present illness by questionnaires responses or by medical records. Well-trained nurses obtained information on parental history of hypertension and lifestyle. Blood pressures (BP) were taken as the average of the second and third measurements. BP categories were defined by the following criteria: normal BP (<120/80 mm Hg), prehypertension (120~139/80~89 mm Hg), and hypertension (>140/90 mm Hg and/or antihypertensive drug user). Cox proportional hazard ratios (HRs) and 95% confidence intervals (CIs) were analyzed after adjusting for age, sex, body mass index, diabetes, hyperlipidemia, smoking, and drinking at baseline.

Results:
In 87,164 person-years of follow-up, 244 incident AF events occurred. The adjusted HRs (95% CIs) of incident AF were 1.56 (1.09~2.24) for hypertension and 1.48 (1.02~2.14) for a maternal history of hypertension. Compared with normal BP and non-maternal history of hypertension, the adjusted HRs (95% CIs) of incident AF were 1.61 (1.10~2.35) for hypertension without a maternal history of hypertension, and 2.29 (1.19~4.40) and 1.95 (1.08~3.50) for prehypertension and hypertension with a maternal history of hypertension, respectively.

Conclusions:
A maternal history of hypertension is a predictor of incident AF. For persons with a maternal history of hypertension, BP control would be under normal BP for AF prevention.
Abstract 370: Stimulation of Angiotensin II Type 2 Receptor Inhibits Binge Eating Disorder with Improving Dopamine Resistance
Authors: Nakaoka Hirotomo , Mogi Masaki , Kan-no Harumi , Tsukuda Kana , Ohshima Kousei , Chisaka Toshiyuki , Wang Xiao-Li , Bai Huiyu , Min Li-Juan , Iwanami Jun , Horiuchi Masatsugu
Published: March 2022

Objective:
Diabetes is associated with abnormal regulation of appetite. Dopaminergic activation in food reward induces binge eating disorder (BED) and leads to increase in food intake. On the other hand, recent studies have demonstrated that stimulation of angiotensin II type 2 (AT
2
) receptor inhibits dopamine (DA) synthesis. In this study, we investigated the relationship between DA level and BED after fasting condition in diabetic mice and the inhibitory effect of AT
2
receptor stimulation on BED and body weight.

Methods:
Adult male type 2 diabetic mice, KKAy, at 10 weeks of age were treated with an intraperitoneal injection of AT
2
receptor agonist, compound 21 (C21), at the dose of 10 μg/kg/day or saline for 2 weeks. DA level in the striatum was measured by microdialysis. Short-term spatial memory and activity were assessed by Y-maze test. After these tests, brain samples were obtained and the expressions of DA receptor D1 (DRD1), DA receptor D2 (DRD2) and DA transporter (DAT) in the substantia nigra were evaluated by immunohistochemical staining.

Results:
Food and water intake and DA level in the striatum were significantly increased 48 hours after fasting compared with non-fasting mice. Administration with C21 significantly attenuated these increase without affecting systolic blood pressure. Treatment with C21 remarkably improved short-term cognitive function and increased explore activity. Moreover, C21 treatment did not affect fasting-induced weight loss, but significantly inhibited rebound weight gain after refeeding compared with vehicle-treated group. In KKAy, the expressions of DRD1, DRD2 and DAT in the substantia nigra were markedly decreased compared with wild-type (C57BL/6J strain) mice, whereas these reductions were improved by administration of C21. Interestingly, we demonstrated that fasting decreased DRD1 and DRD2 expression and increased DAT; however, administration of C21 inhibited these reductions of DRD1 and DRD2.

Conclusion:
Activation of AT
2
receptor could contribute to inhibition of BED and rebound weight gain with modulation of dopamine signaling. These results indicate that stimulation of AT
2
receptor may be a new therapeutic approach to improve the eating disorder associated with dopamine resistance.
Abstract 373: Mineralocorticoid Receptor and Rhoa/rho Kinase Contribute to Vascular Dysfunction in Obese Db/db Mice
Authors: Nguyen Dinh Cat Aurelie , Callera Glaucia E , Antunes Tayze T , He Ying , Montezano Augusto C , Bruder-Nascimento Thiago , Touyz Rhian M
Published: March 2022

Increasing evidence indicates that adipose tissue modulates vascular function. Our previous studies demonstrated that adipocytes secrete aldosterone. We hypothesized that adipocyte-releasing factors induce vascular dysfunction through mineralocorticoid receptor (MR)-dependent mechanisms. This study explored the small G protein RhoA/Rho kinase as a pathway linking aldosterone/MR and the paracrine effects of adipose tissue in the vasculature, since the activation of this pathway regulates vascular contraction. Diabetic obese mice (db/db) and their control counterparts (db/+) were treated with MR antagonist (MRA) (K canrenoate, 30mg/Kg/day, S.C., 4 weeks) or RhoA/Rho kinase inhibitor, fasudil (30 mg/Kg/day, S.C., 3 weeks). Blood pressure levels were similar between groups. Fasudil, but not MRA, reduced plasma glucose levels in db/db mice (db/+:12.3±6; db/db: 30.1±3; MRA-treated db/db: 32.5±2; fasudil treated db/db: 17.1±2; mmoL/mL). Arteries from db/db mice displayed reduced relaxation to 10
-
5
M acetycholine (Ach; db/+:79.4±3.9% vs db/db: 14.3±3.1%). Arteries from MRA-treated db/db mice exposed to fat conditioned medium (FCM) had improved responses to Ach (MRA-treated db/db: 28.3±1.9% vs MRA-treated db/db +FCM: 42.8±4.3%). Increased norepinephrine (NE)-induced contraction was observed in db/db mice in endothelium-denuded arteries (10
-5
M NE; db/+: 1.5 ± 0.1 vs db/db: 2.2 ± 0.1; mN/mm). NE responses were reduced in MRA-treated db/db mice (1.7±0.1 mN/mm) or fasudil (1.6±0.2 mN/mm). Vascular calcium sensitivity was similar between depolarized arteries from db/+ and db/db. Fasudil treatment reduced calcium-induced contraction in both groups with greater effect in db/db mice (5 mmol/L CaCl
2
; db/+: 73 % vs db/db: 57 %). MR blockade did not affect vascular RhoA/Rho kinase activity (RhoA membrane to cytosol translocation) in db/db mice. Our data suggest that in db/db mice: MR and RhoA/Rho kinase signaling contribute to adipose modulation of vascular/contraction. While MR plays a role in vascular relaxation, RhoA/Rho kinase signalling is important in contractile dysfunction, adipocyte secretion, and glucose metabolism - processes that may contribute to cardiovascular injury in obesity-associated diabetes.
Abstract 430: Blood Pressure Monitoring in the Biological Time Structures and in Pre- /Post-Surgery Seasonal Variations
Authors: Lee Mary S , Lee John S , Halberg Franz , Lee Jong Y
Published: March 2022

Biological time structures (clock-hour), including yearly rhythm (circannual), ~weekly (circaseptan) and circadian are considered essential parameters in blood pressure (BP) to recognize and treat any risk elevation as well as overt disease. BP and heart rate (HR) in adolescents and adults were monitored automatically around the clock at 30-minute to hourly intervals for seasons and/or years. Data was analyzed by the linear least square rhythmometry. All showed significant circadian fluctuations in BP and HR. BP was about 10-20 mm Hg higher in the adult group while HR was lowered in average about 10 beats/min. The circadian response peaked in the noon to the late afternoon hours: SBP: 18:08; DBP: 17:08; HR: 17:40 with the impression from the 5 adult subjects, individual differences notwithstanding, while one teen showed shift changes to the early morning hours: SBP: 01:48; DBP: 01:24: HR: 00:12. Circannual BP and HR revealed significant rhythms with peaks on October 27 (SBP), November 26 (DBP) and October 31 (HR). While the winter month BP and HR were the lowest in comparison to the other 3 seasons, further sudden drops of both BP and HR were noted in the individual at the point of surgery in mid-January. However, when the surgery performed in the same individual in November, the drop was minimal, while early spring time surgery increased SBP significantly with a minimal increase in DBP and HR. Sharp increased double amplitudes (Amps) (SBP 73; DBP 35; HR 24, all P<0.0001) observed in the winter season. The circaseptan double Amps in the adolescent were 13 ± 4 (P=0.019) for SBP and 8 ± 3 mm Hg (P=0.030) for DBP, with acrophases occurring on late Sunday, early Monday (at -50° and -67° from 00:00 on Saturday to Sunday for SBP and DBP, respectively, with 360° ≡ 1 week). Post-OP Amps were significantly shallow compared with control: SBP: 45 vs. 19; DBP: 21 vs. 6; HR: 21 vs. 12 mm Hg, while acrophase (peak hrs) was various: pre-/post-op SBP: 16:44 vs. 11:32; DBP: 11:44 vs. 01:16; HR: 11:44 vs. 13:08 hrs. The increased or shallow Amp can be a predictable risk factor of CVD. Circaseptan acrophases coincide with the time of increased vascular morbidity/mortality in our studies. Chronomes broader than circadian should be considered in interpreting BP responses as a gauge of vascular disease status.
Abstract 448: Assessment of Drug Interaction Potential Between LCZ696 and Warfarin
Authors: Ayalasomayajula Surya , Jordaan Pierre , Pal Parasar , Albrecht Diego , Langenickel Thomas , Sunkara Gangadhar , Chandra Priyamvada
Published: March 2022

Objective:
LCZ696 is a first-in-class angiotensin receptor neprilysin inhibitor (ARNI) being developed for the treatment of cardiovascular diseases including hypertension and heart failure. Ingestion of LCZ696 results in systemic exposure to AHU377 (an inactive prodrug converted to LBQ657, neprilysin inhibitor) and valsartan (angiotensin receptor inhibitor). Warfarin, a CYP2C9 substrate and narrow therapeutic index drug , is administered to patients with cardiovascular diseases including heart failure . Since LCZ696 is a known weak inhibitor of CYP2C9, the objective of this study was to determine whether LCZ696 affects the pharmacokinetics (PK) and pharmacodynamics (PD) of warfarin. Also, the warfarin effect on the PK of LCZ696 was evaluated.

Methods:
This study employed an open label, single blind, two-period, crossover design in 26 healthy subjects. In each period, subjects received either 200 mg LCZ696 or matching placebo b.i.d for 10 days with a single dose of 25 mg warfarin co-administered on Day 5. Serial PK samples were collected on Day 4 and Day 5 and analyzed using validated LCMS/MS methods. Prothrombin time (PTT) and INR values were measured for up to 144 hours post warfarin dose. The PK parameters including Cmax and AUCs of LCZ696 analytes (LBQ657 and valsartan) and warfarin (R-warfarin and S-warfarin) and PD parameters of warfarin (peak PTT, peak INR, AUC-PTT, and AUC-INR) were determined using noncompartmental methods and results were statistically evaluated.

Results:
The 90% confidence intervals (CI) of geometric mean ratios of Cmax and AUCs (test/reference) for R-warfarin, S-warfarin, LBQ657, and valsartan were within the 80 - 125% suggesting lack of PK interaction between LCZ696 and warfarin. Further, the 90% CI of geometric mean ratios of average (AUC effect/144) and peak PD effects (PTT and INR) were also within the 80 - 125% range indicating no impact of LCZ696 on warfarin PD.

Conclusion:
Co-administration of 200 mg LCZ696 b.i.d with single dose of 25 mg warfarin did not alter the PK exposures of LCZ696 analytes (LBQ657 and valsartan) and warfarin. Also, LCZ696 did not affect the PD effects of warfarin. LCZ696 200 mg b.i.d was safe and well tolerated when administered alone or with single dose of 25 mg warfarin.
Abstract 45: Reduced Vascular Remodeling and Improved Endothelial Function in Transglutaminase 2 Knock-Out Mice Treated with Angiotensin II
Authors: Savoia Carmine , Arrabito Emanuele , Sada Lidia , Michelini Serena , Pucci Lorenzo , Briani Martina , Nicoletti Carmine , Candi Eleonora , Schiffrin Ernesto L , Volpe Massimo
Published: March 2022

Transglutaminase 2 (TG2) may modulate cell-matrix interactions by inducing cross-linking of proteins. We previously demonstrated that angiotensin II (Ang II) positively regulated TG2 expression in vascular smooth muscle cells from SHR. Here we hypothesized that Ang II induces vascular remodeling in part through TG2.

TG2-knockout mice (TG2-K/O, 8 weeks old, 6 for each group) and age-matched wild type (WT) control mice were treated or not with Ang II (400 ng/kg/min) for 14 days. Blood pressure (BP) was measured by tail-cuff method. Endothelium-dependent and -independent relaxation were assessed by concentration-response curves to acetylcholine (1 nM to 100 μM) ± L-NAME (100 μM) and sodium nitroprusside (10 nM to 1 mM) respectively, in mesenteric arteries pre-contracted with norepinephrine (10 μM). Media-to-lumen ratio (M/L) and cross sectional area (CSA) were evaluated on pressurized preparations.

BP was higher in TG2-K/O mice compared to WT (120.3±1.3 mmHg vs 88.3±1.9 mmHg, P<0.05), Ang II infusion significantly increased BP only in WT (+28% vs untreated WT, P<0.05), whereas BP was unchanged in TG2-K/O after Ang II infusion. Endothelium-dependent relaxation was similarly preserved in untreated WT, TG2-K/O and Ang II-treated TG2-K/O. Ang II infusion impaired acetylcholine-induced relaxation only in WT (-50% vs untreated WT, P<0.05). L-NAME blunted acetylcholine-induced relaxation in all the groups except in Ang II-treated WT, suggesting an impairment of NO production only in this group. Endothelium-independent relaxation was similar in all groups. TG2-K/O presented reduced M/L as compared to WT (4.8±0.3% vs 6.5±0.2%, P<0.05). Ang II infusion increased M/L only in WT (+13% vs untreated WT, P<0.05). M/L resulted unchanged in TG2-K/O after Ang II infusion. CSA was similar in all groups.

In conclusion, despite the higher BP values, TG2-K/O presented improved vascular remodeling compared to WT. Ang II failed to increase M/L and impair endothelial function in TG2-K/O. Hence TG2 may play a role in Ang II-induced vascular structural and functional alterations.
Abstract 451: Central Hemodynamics Constitutes the Link Between Microalbuminuria and Elevated B-type Natriuretic Peptide Level: Implication for Cardio-renal Syndrome in Hypertension
Authors: Hashimoto Junichiro , Ito Sadayoshi
Published: March 2022

Recent research has noted a pathological link between the heart and kidney, known as cardio-renal syndrome. However, little is clear about the pathophysiological mechanism behind it. A relationship of altered central hemodynamics to microalbuminuria in hypertension has been demonstrated (
Hypertension
2011;58:839-846). The present study was conducted to investigate two hypotheses: 1) a link between albuminuria and plasma B-type natriuretic peptide (BNP); and 2) involvement of central hemodynamics in this link. In 392 patients with uncomplicated hypertension (age, 56±12 years), the radial pressure waveforms were recorded with applanation tonometry to estimate the central aortic pressure parameters. The pulse wave velocity (PWV) was measured in carotid-femoral (aortic) and carotid-radial (peripheral) regions. Albuminuria was defined as urinary albumin/creatinine ratio (UACR) ≥30 mg/g of creatinine. The log-transformed BNP (median, 14 pg/ml) was correlated (
P
≤0.006) with aortic systolic (
r
=0.31) and pulse (
r
=0.46) pressures, aortic forward (
r
=0.41) and augmented (
r
=0.26) pressures, mean arterial pressure (
r
=0.14), and aortic (
r
=0.39) but not peripheral (
r
=–0.09) PWV. The correlation with the aortic pulse pressure tended to be closer than the brachial pulse pressure (
P
=0.07). The BNP was correlated (
P
<0.001) positively with UACR (
r
=0.34), and negatively with estimated glomerular filtration rate (eGFR,
r
=–0.39). The presence of albuminuria was associated with an elevation of BNP (≥40 pg/ml) in a stepwise logistic model adjusted for age, body mass index, diabetes, eGFR, use of renin-angiotensin inhibitors and β-blockers (odds ratio: 2.74;
P
=0.007). Of note, however, additional adjustment for the aortic pulse pressure rendered this association statistically insignificant, and the aortic pulse pressure emerged as the strongest predictor of the BNP elevation (odds ratio: 1.41 per 10mmHg increase;
P
=0.003). In each patient subgroup with normo-, micro-, and macro-albuminuria, wider aortic pulse pressure was always associated with higher BNP level. In conclusion, albuminuria and elevated BNP are closely connected in hypertension. The central hemodynamics plays a pivotal role in mediating this cardio-renal connection.
Abstract 462: Gene-targeting of Npr1 Reveals the Retinoic Acid-mediated Enhanced Expression of Guanylyl Cyclase-A/Natriuretic Peptide Receptor-A and Attenuation of Renal Fibrosis in Mutant Mice
Authors: Kumar Prerna , Das Subhankar , Pandey Kailash N
Published: March 2022

Mice lacking functional
Npr1
gene (coding for guanylyl cyclase/natriuretic peptide receptor-A, GC-A/NPRA) exhibit hypertension and kidney disease, however, the mechanisms regulating
Npr1
expression are not well understood. The objective of the present study was to determine the effect of all-
trans
retinoic acid (ATRA) on
Npr1
gene expression and reduction of renal fibrosis in gene-targeted mutant mice. Adult (24-week old) male
Npr1
gene-disrupted heterozygous (1-copy; +/-), wild-type (2-copy; +/+), and gene-duplicated (3-copy; ++/+) mice were treated by injecting ATRA (0.5 mg/Kg/day) intraperitoneally for 2-weeks. After treatment with ATRA, the renal
Npr1
mRNA levels was increased in 1-copy mice by 4.6-fold, (1.4 ± 0.05 vs. control, 0.3 ± 0.02; p < 0.05), 2-copy mice by 3-fold, (3.7 ± 0.2 vs. control 1.2 ± 0.1; p < 0.05), and 3-copy mice by 2-fold (3.2 ± 0.3 vs. control 1.6 ± 0.2; p < 0.05). The retinoic acid increased renal cGMP levels (pmol/mg protein) in 1-copy (28.9 ± 1.1 vs. control, 7.1 ± 0.4; p < 0.05), 2-copy (55.8 ± 1.9 vs. control 22.6 ± 1.8; p < 0.05), and 3-copy (85.8 ± 4.3 vs. control 47.8 ± 3.2; p < 0.05) mice. A significant decrease in systolic blood pressure (SBP) was observed in ATRA-treated 1-copy mice (114.6 ± 2.2, p < 0.01) vs. 1-copy control mice (128.2 ± 2.1). Higher tubulo-interstitial fibrosis present in 1-copy mice was attenuated after treatment with ATRA (35%, p < 0.05). Immunohistochemistry and Western blot analyses exhibited the reduction in renal expression of α-SMA (25%, p < 0.05 and 35%, p < 0.05, respectively) in ATRA-treated 1-copy mice compared with 1-copy control mice. Interestingly, ATRA significantly enhanced the levels of immunosuppressive cytokine IL-10 (pg/mg protein) in the kidneys of 1-copy (43.3 ± 3.1 vs. control 23.4 ± 1.8; p < 0.05), 2-copy (67.1 ± 5.5 vs. control 56.5 ± 3.7; p < 0.05), and 3-copy (102.7 ± 6.7 vs. control 81.4 ± 3.2; p < 0.05) mice. The results show that ATRA induces
Npr1
gene expression and function, lowers SBP, and attenuates renal fibrosis in 1-copy
Npr1
mice. The findings of this study indicate that ATRA can serve as a possible therapeutic agent in attenuation of renal fibrosis under reduced
Npr1
gene copy number, which will have important implications in prevention of hypertension and renal remodeling process.
Abstract 472: Intrarenal Ghrelin Receptor Blockade Induces Natriuresis and Diuresis in SHR
Authors: Kemp Brandon A , Howell Nancy L , Padia Shetal H
Published: March 2022

Ghrelin, a novel 28 amino-acid peptide hormone, was originally discovered as a centrally-acting orexigenic hormone. Our laboratory has recently demonstrated that ghrelin receptors are present in the kidney in collecting duct cells where they increase ENaC-dependent Na+ reabsorption in normal rats. However, nothing is known about the function of intrarenal ghrelin receptors in SHR. To evaluate this system, uninephrectomized 10-week-old female SHR (N=16) received three cumulative 1 h renal interstitial (RI) infusions of the specific ghrelin receptor antagonist D-LYS-GHRP-6 (4, 6, 8 μg/min; Tocris) following a 1 h control infusion of vehicle 5% dextrose in water (D5W). Rats that received 4 cumulative 1 h RI infusions of D5W served as time controls. Mean arterial pressure values, urine Na+ excretion rates (UNaV), and urine flow rates were determined for each period. Following the RI infusion of D-LYS-GHRP-6, UNaV increased from a baseline value of 0.035 ± 0.004 μmol/min to 0.062 ± 0.011 μmol/min in period 1 (P<0.05), 0.093 ± 0.018 μmol/min in period 2 (P<0.05), and 0.13 ± 0.019 μmol/min in period 3 (P<0.01). Furthermore, RI D-LYS-GHRP-6 infusion increased urine flow rate from a baseline value of 0.002 ± 0.0003 ml/min to 0.005 ± 0.001 ml/min during period 1, 0.006 ± 0.002 ml/min during period 2, and 0.01 ± 0.003 ml/min during period 3 (P<0.05 for all 3 periods). There were no significant changes in either UNaV or urine flow rate in the time controls. Mean arterial pressure responses remained unchanged following the RI infusions, suggesting that all agents were confined to the kidney. These data introduce a potential new therapeutic target for the treatment of sodium and water retention in SHR.
Abstract 474: High Salt Diet Modulates Ca
2
+-Sensing Receptor and Cytochrome P450 Isoform Expression in Mesenteric Arteries in Dahl Salt-Sensitive Rats
Authors: Ayanwale Lydia D , Williams Cicely L , Awumey Emmanuel M
Published: March 2022

The Ca
2+
-sensing receptor (CaSR) mediates relaxation of isolated, phenylephrine-contracted mesenteric arteries, which express the receptor in the sensory network. We tested the hypothesis that high salt alters the expression of CaSR and its CYP enzymes downstream of the receptor in salt-sensitive hypertension. Dahl salt−sensitive (DS) and Dahl salt-resistant (DR) rats were placed on an 8% NaCl diet (HS) for 1, 2, and 4 weeks. Using tail cuff plethysmyography, blood pressures were measured before and after HS diet treatment. Our results show a significant increase in blood pressure in DS animals on HS diet (Control
:

116 ± 4.8 mmHg; 1 week HS:

164.4 ± 8.7 mmHg; 2 weeks HS: 175.1 ± 7.2 mmHg;
p < 0.05
, ANOVA). Western blot analysis of proteins extracted from mesenteric arteries showed an increase in CaSR, CYP1A1, and CYP2J2 expression in DS rats on HS diet, but not in arteries from DR rats on the same diet. The data show that HS increased mean arterial blood pressure, as expected, and increased the expression of the mature form of the CaSR while decreasing expression of the immature form. Increased CYP isoform expression was observed as well, however, these changes did not reduce blood pressure. These new findings suggest an abnormality in CaSR activity in salt −sensitive hypertension, which may provide a molecular target for anti-hypertensive therapies.
Abstract 499: Renal Endocannabinoids in Salt-Sensitive Hypertension of SHR
Authors: Varagic Jasmina , Gallagher Patricia E , Grabenauer Megan , Thomas Brian F
Published: March 2022

Recent evidence suggests that endocannabinoids (ECs) acting on two major cannabinoid receptors (CB
1
and CB
2
) regulate blood pressure and cardiovascular and renal function in health and disease. However, there is no evidence of the role of ECs in the salt-induced renal damage in hypertensive subjects. In the present study we investigated changes in the content of two major ECs, 2-arachidonoylglycerol (2-AG) and anandamide (
N
-arachidonoylethanolamine), and related functional changes in the kidneys of spontaneously hypertensive rats (SHR) in response to increased dietary salt intake. Male SHRs were given an 8% salt diet (HS; n=7), while their age-matched controls (C; n=7) received standard chow for 5 weeks. ECs were measured by UPLC/MS-MS methods on Applied Biosystems Mass Spectrometry Systems. After 5 weeks on a high salt diet, renal 2-AG (4.29 ± 0.40 vs. 3.32 ± 0.17 ng/mg; p<0.05) but not anandamide (2.12 ± 0.17 vs. 2.27 ± 0.07 pg/mg) increased in the salt-loaded SHR. Dietary salt excess increased mean arterial pressure (203 ± 5 vs. 171 ± 7 mmHg; p<0.05), urinary protein excretion (159 ± 37 vs. 22 ± 2 mg/day), and significantly decreased renal blood flow measured by flow probe (2.93 ± 0.55 vs. 6.67 ± 0.56 ml/min/mg; p<0.05) and glomerular filtration rate assessed by creatinine clearance (1.15 ± 0.14 vs. 1.77 ± 0.08 ml/min; p<0.05). Our previous study showed a reduction in CB
1
receptor mRNA while CB
2
receptor gene expression was increased in the kidney of salt-loaded SHR. Together these results suggest that alterations in renal ECs may play a significant role in the development of renal injury due to high dietary salt intake.
Abstract 5: Intrarenal Responses to Angiotensin I and Angiotensin (1-7) in the Stroke-Prone Spontaneously Hypertensive Rat Are Y Chromosome Dependent
Authors: Sampson Amanda K , Andrews Karen L , Graham Delyth , McBride Martin W , Chin-Dusting Jaye P , Dominiczak Anna , Jennings Garry L
Published: March 2022

The Y chromosome accounts for 15-20mmHg difference in arterial pressure; the gene(s) and mechanism(s) underlying this effect remain unknown. One candidate gene, the Sry3 gene, expressed exclusively on the hypertensive SHR Y chromosome has been shown
in vitro
to interact with the renin angiotensin system (RAS). Using a reciprocal Y consomic rat approach, we investigated the functional consequences of this interaction
in vivo
; examining the interaction of the Y chromosome with the renal vasculature. 16 week old normotensive rats (WKY), hypertensive rats (SHRSP) and two reciprocal Y consomic rat strains; one comprising the WKY autosomes and X chromosome with the hypertensive Y chromosome (WKY.SP
Gla
Y) and
vice versa
(SP.WKY
Gla
Y) were examined. We confirmed via radiotelemetry that systolic blood pressure (SBP) was Y chromosome dependent; SP.WKY
Gla
Y had lower SBP than SHRSP (195±5mmHg vs 227±8mmHg, P<0.03) and was higher in WKY.SP
Gla
Y compared to WKY (157±3mmHg vs 148±3mmHg, P<0.05). Compared to WKY rats, the vasodilator arm of the RAS was enhanced in SHRSP as evidenced by a higher plasma Ang(1-7):Ang II ratio (WKY:0.13±0.01 vs SHRSP:1.33±0.4, P<0.005) and a blunted renal blood flow (RBF) response to graded intrarenal Ang I and Ang(1-7) infusions. In response to 10ng/kg we observed a reduction in RBF of WKY:58±6% and 16±6% vs SHRSP: 17±6%, P<0.01 and 1±4%, P<0.05 respectively. Introgression of the normotensive Y chromosome into the SHRSP background (SP.WKY
Gla
Y) resulted in a reduction in plasma Ang(1-7):AngII ratio (SP.WKY
Gla
Y: 0.24±0.02, P<0.01) and an increased vasoconstrictor response to intrarenal Ang I and Ang(1-7) infusion. In response to 10ng/kg bolus RBF in SP.WKY
Gla
Y reduced by 45±14%, P<0.01 and 41±18%, P<0.005, respectively compared to SHRSP, demonstrating the blunted responsiveness in the SHRSP is Y chromosome dependent. Investigation of the interlobular arteries via wire myography revealed increased sensitivity to Ang II in SHRSP compared to WKY which was reduced following introgression of the WKY Y chromosome (SP.WKY
Gla
Y) confirming that the Y chromosome influences the RAS in the renal vasculature. This study provides novel evidence that the Y chromosome influences the vasodilatory arm of the RAS and intrarenal vascular function.
Abstract 513: Impact of Sex and Race on Angiotensin (1-7) in Humans
Authors: Harris Ryan A , Zimmerman Margaret A , Seigler Nichole , Sullivan Jennifer C
Published: March 2022

Approximately 30% of American adults are diagnosed with hypertension; however, only ~50% achieve adequate blood pressure (BP) control with current therapeutic options. Preclinical data demonstrate that a sexual dimorphism exists for angiotensin 1-7 (Ang (1-7)), a vasodilatory peptide that has therapeutic promise in controlling BP. This study sought to determine if 1) sex and race differences in Ang (1-7) are present in normotensives adults, and 2) increases in Ang (1-7) are related to lower BP in hypertensive adults. Blood samples, BP, and flow-mediated dilation (FMD, n=29) were assessed in 45 adults (52% women, 44% African American (AA)). Systolic BP was lower (p=0.013) in women compared to men (111±4 mm Hg vs. 126±4 mm Hg) and higher (p=0.024) in AA compared to Caucasians (C; 125±4 mm Hg vs. 111±4 mm Hg). Diastolic BP was higher (p=0.021) in AA compared to C (77±3 mm Hg vs. 65±4 mm Hg). Overall, Ang (1-7) was highest in C women (44±5 pg/ml) followed by AA women (38±5 pg/ml), C men (33±4 pg/ml), and AA men (30±6 pg/ml). Only within C was a significant correlation between FMD and Ang (1-7) observed (r=0.573;p=0.041). Overall, the change in Ang (1-7) following treatment with candesartan (n=6) was positively associated with the change in FMD (r=0.485) and inversely associated with the change in systolic BP (r=-0.431). These data are the first to identify, in humans, the existence of a sexual and racial dimorphism in Ang (1-7). Our findings support the therapeutic potential of Ang (1-7) to reduce BP and improve endothelial function in hypertensive adults.
Abstract 523: Differences in Sodium and Potassium Regulation during Mental Stress in Normotensive Youths
Authors: Stewart Deborah L , Harshfield Gregory A , Dong Yanbin , Harris Ryan A , Kapuku Gaston , Grim Clarence E
Published: March 2022

Background:
In Dahl salt sensitive rats, the “air puff stress” increases blood pressure (BP) and is associated with acute renal sodium (Na) retention. This study revealed increased dietary potassium (K) may abolish stress induced Na retention. Our human studies have shown the mental stress (MS) of playing a video game increases BP in youths, but the effect on Na excretion is bimodal. Retainers (NaR), retain Na with MS while excreters (NaE), excrete Na during MS. A greater increase in BP with a prolonged effect is seen in NaR. This stress induced NaR phenomena can be abolished by Ang II blockage. Because BP effects of Na intake are closely related to K intake, this study sought to compare U
Na
V and U
K
V before and after MS between NaE and NaR.

Methods:
We compared parallel changes in U
Na
V, U
K
V, aldosterone (aldo), and Ang II in response to 1 hour of MS in 90 healthy youths (16-19 years) after a 3 day Na/K controlled diet. We compared NaE (n=64) to NaR (n=26).

Results:
The change in U
Na
V (5.5mEq±0.6SEM vs. -3.4mEq±0.7SEM; p=<0.001), U
K
V (2.1mEq±0.3SEM vs. 0.4mEq±0.2SEM; p=0.002), aldo (-15.4 pg/mL ±5.3SEM vs. 3.7 pg/mL ±7.9SEM; p=0.055) and Ang II (0.4 pg/mL ±0.7SEM vs. 4.9pg/mL±3.6SEM; p=0.085) was greater in NaE vs NaR, as seen in the figure. Similarly, both groups had a baseline U
Na
V/U
K
V ratio of 4:1, which changed during stress to 2:1 in NaR and 3:1 in NaE.

Conclusion:
MS in normotensive youths has remarkably different effects on U
Na
V and U
K
V. Salt sensitive animal models have shown K supplementation may reverse Na retention in response to MS. This study suggests K supplementation may reverse the Na/K ratio to pre-stress conditions. Figure: Delta value comparison between NaE & NaR in UNaV, UKV, aldo, & Ang II.
Abstract 536: Resiniferatoxin-induced Disorders in Glucose Homeostasis and Hypertension: Role of Trpv1-positive Sensory Nerves
Authors: Wang Donna H , Yu Shuang-Quan
Published: March 2022

Resiniferatoxin (RTX), an ultrapotent agonist of the transient receptor potential vanilloid 1 (TRPV1) channels, has been used to relieve pain in humans in clinical trials that are ongoing, completed, or terminated due to its unwanted side effects that have not been well understood. We test the hypothesis that RTX causes disorders in glucose homeostasis, renal functional and structural impairment, and hypertension via insulin receptor-mediated increases in sympathetic nerve activity. Male Wistar rats were injected with RTX (0.2 mg/kg, i.p.) or vehicle. Pancreatic and renal TRPV1 levels were decreased 4 weeks after RTX treatment, confirming effective and lasting treatment of RTX (p<0.05). RTX decreased plasma insulin levels (Control: 1.4±0.1 vs RTX: 0.8±0.1 ng/ml, p<0.05) but improved glucose tolerance (p<0.05). RTX increased the translocation of skeletal muscle glucose transporter 4 (Glut4) from cytosol to cell membrane indicated by increased membrane fraction but decreased cytosol fraction of Glut4 in RTX-treated rats (p<0.05). RTX increased levels of phosphorylated insulin receptor substrate 1 in sympathetic ganglia (Control: 0.43±0.04 vs RTX: 0.65±0.08, p<0.05), renal tyrosine hydroxylase contents, and urinary norepinephrine concentrations (Control: 352±70 vs RTX: 781±142 ng/day, p<0.05). When TRPV1 expressed in sensory neurons and their nerve endings innervating the kidney was knocked down by intrathecal (i.t) injection of TRPV1 shRNA, insulin (3nmol/kg, i.t.)-induced increases in efferent renal nerve activity were further enhanced (Control: 40±3 vs TRPV1 shRNA: 53±2, p<0.05). RTX decreased creatinine clearance (Control: 0.58±0.04 vs RTX: 0.46±0.04 ml/min/100 gbwt, p<0.05) and increased levels of plasma urea, urinary albumin, renal connective tissue growth factor, and systolic blood pressure (Control: 128±1 vs RTX: 143±2 mmHg, p<0.05). Thus, RTX treatment causes disorders in glucose homeostasis, renal functional and structural impairment, and hypertension, possibly via increasing sympathetic nerve activity resulting from enhanced stimulatory effects of the insulin receptors expressed in sympathetic nerves and weakened inhibitory effects of TRPV1-positive sensory nerves on sympathetic nerves.
Abstract 537: Segment-specific Intrathecal Injection of Metformin Protects Against High Fat Diet-induced Impairment in Afferent Renal Nerves and Renal Function
Authors: Wang Donna H , Yu Shuang-Quan
Published: March 2022

Metformin (Met), a drug for treating type 2 diabetes by suppressing glucose production and increasing insulin sensitivity, may possess neuroprotective and neurogenesis effects. We test the hypotheses 1) that high fat diet (HFD) intake impairs afferent renal nerves expressing transient receptor potential vanilloid 1 (TRPV1) channels, leading to increased efferent renal nerve activity (ERNA), decreased renal function, and hypertension; and 2) that Met given intrathecally (i.t.) to segments (T8-L3) supplying the kidney prevents HFD-induced impairment in afferent renal nerves, renal function, and hypertension. Met (1 ng/kg, i.t. daily) or vehicle was given to rats fed a HFD or normal fat diet (Con) for 8 weeks. Glucose or insulin tolerance was impaired to the same degree between rats fed HFD with and without Met treatment. HFD decreased renal TRPV1 expression, plasma levels of calcitonin gene-related peptide, and afferent renal nerve activity (ARNA, Con: 126±11, Met: 136±14, HFD: 77±7, HFD+Met: 121±8, p<0.05) in response to intra-pelvis perfusion of capsaicin (a TRPV1 agonist at 4 mM, 3 min, 20 ml/min), which were prevented by Met. HFD increased renal tyrosine hydroxylase contents, ERNA responses to intrathecal injection of muscimol (a GABA-A receptor agonist at 3 nmol/kg), and urinary norepinephrine levels, which were prevented by Met. HFD decreased creatinine clearance (Con: 0.40±0.04, Met: 0.52±0.07, HFD: 0.12±0.05, HFD+Met: 0.30±0.04 ml/min/100 gbwt, p<0.05) and urinary Na+ excretion, and increased levels of urinary albumin, plasma urea, renal collagen deposition, renal connective tissue growth factor, and systolic blood pressure (Con: 132±1, Met: 130±1, HFD: 145±3, HFD+Met: 132±1 mmHg, p<0.05), which were prevented or attenuated by Met. Thus, HFD impairs TRPV1-positive afferent renal nerves and renal function, and increases renal sympathetic nerve activity and blood pressure. Segment-specific intrathecal injection of Met protects against HFD-induced impairment in afferent renal nerves and renal function, and prevents HFD-induced hypertension.
Abstract 565: Angiotensin Ii Type 1 Receptor Activation is Required for Angiotensin Ii and Interleukin 6-induced Augmentation of Angiotensinogen Expression in Mouse Renal Proximal Tubular Cells
Authors: Satou Ryousuke , Hopfer Ulrich , Navar L G
Published: March 2022

In angiotensin II (AngII) dependent hypertension, an inappropriate elevation of intrarenal Ang II leads to the progression of hypertension and kidney injury. Intrarenal angiotensinogen (AGT), produced mainly by renal proximal tubular cells (PTC), is a crucial factor in regulation of intratubular Ang II. Increases in intrarenal AGT are associated with increases in intrarenal immune cells and interleukin 6 (IL-6) levels which contribute to Ang II-dependent AGT augmentation. These results suggest that Ang II and IL-6 synergize to increase AGT levels during progression of Ang II-dependent hypertension. However, the interactions among Ang II, Ang II type 1 receptors (AT1R) and IL-6 on AGT expression and the mechanisms have not been delineated. In this study, mouse PTC (WT-PTC) and AT1aR knockout strains (AT1aRKO-PTC) were used to test the existence of this synergism between Ang II and IL-6. Basal AGT mRNA expression levels in PTC were determined and compared with that in mouse mesangial cells which is another established AGT-expressing kidney cells. AGT mRNA expression levels were measured by real-time RT-PCR and normalized based on GAPDH levels. WT-PTC had 21.1-fold (± 2.0) higher basal AGT mRNA levels than mesangial cells, indicating markedly high AGT production by the PTC. Further, WT-PTC and AT1aRKO-PTC were incubated with 100 nM Ang II and/or 400 nM IL-6 for 24 h. Stimulation with either Ang II or IL-6 alone did not significantly alter AGT mRNA expression in both WT-PTC and AT1RaKO-PTC. Co-stimulation with Ang II and IL-6 increased AGT mRNA levels (1.59 ± 0.04, ratio to control) in WT-, but not AT1aRKO-PTC. To test for involvement of the NF-κB signaling pathway in the synergism between Ang II and IL-6, activation of NF-κB as an AT1R signaling pathway was evaluated by western blot analysis. Furthermore, WT-PCT were treated with an NF-κB inhibitor during the co-stimulation. In WT-PTC, IκB levels were decreased by the co-stimulation (0.61 ± 0.04, ratio to control) indicating NF-κB activation. Ten μM Parthenolide, an NF-κB inhibitor, attenuated the AGT mRNA increase induced by the co-stimulation. These results suggest that simultaneous AT1R activation acts synergistically with IL-6 to increase AGT expression in mouse PTC and that the signaling pathway involves NF-κB.
Abstract 601: Bcl10 Deficiency of Both Cardiac Cells and Bone Marrow Derived Cells Attenuates Angiotensin (Ang) II-induced Target Organ Damage
Authors: Marko Lajos , Henke Norbert , Park Joon-Keun , Spallek Bastian , Qadri Fatimunnisa , Apel Ingrid J , Oravecz-Wilson Kay , Przybyl Lukasz , Luft Friedrich C , Dechend Ralf , Ruland Jürgen , Lucas Peter C , McAllister-Lucas Linda M , Mueller Dominik N
Published: March 2022

Background
Bcl10 is a member of the CARMA3-Bcl10-MALT1 (CBM) signalosome that links angiotensin (Ang) II and NF-κB signaling. We have shown earlier that Ang II-infused Bcl10 knockout (KO) mice develop less inflammatory cell infiltration and less fibrosis coupled with improved arrhythmogenic electrical remodeling compared to wild-type (WT) littermates. In our present study we investigated the mechanism behind this phenotype.

Methods and Results
Bone marrow (BM) from WT and green fluorescence protein transgenic WT mice (GFP+ WT) was transferred into WT and Bcl10 KO mice. Conversely, Bcl10 KO BM was transferred into WT mice. After BM regeneration mice were uni-nephrectomized, 1% NaCl was given in the drinking water and received Ang II (1.44 mg/kg/d) for 14 days. Both Bcl10 BM transplanted WT mice and WT BM transplanted Bcl10 KO mice showed less immune cell infiltration compared to WT BM transplanted WT mice (macrophages: 44.8±2.7 and 46.8±1.8 vs. 62.0±1.5; CD4+ T-cells: 22.8±1.4 and 24.3±1.3 vs. 31.2±0.9; CD8+ T cells 24.2±1.2 and 28.0±1.1 vs. 39.5±1.1 cells/heart section, respectively) despite similar cardiac hypertrophy. In parallel with cell infiltration these mice developed less perivascular collagen 1 and interstitial fibronectin deposition, as well. Immunohistological and qPCR analysis revealed less VCAM-1, ICAM-1 and chemokines (MCP-1 and RANTES) expression in WT BM transplanted Bcl10 KO mice compared to Bcl10 BM transplanted or WT BM transplanted WT mice. Moreover, in vitro endothelial cells transfected with Bcl10 siRNA showed 5-fold less increase in monocyte attachment after Ang II treatment as endothelial cells transfected with control siRNA. Additionally, transplantation with GFP+ WT BM cells revealed that Bcl10 KO mice recruit less BM-derived GFP+/FSP1+ fibroblasts compared to GFP+ WT BM transplanted WT mice (51.3±1.3 vs. 23.5± 1.3 cells/heart section) suggesting that non-cardiac cells contribute to fibrosis too. Finally, intraperitoneal injection of MCP-1 into WT mice significantly increased the number of peritoneal CD11b+ cells, whereas Bcl10 KO mice showed no increase in response to MCP-1.

Conclusion
Our results demonstrate that cardiac fibrosis in Bcl10 KO mice depends on both immune/BM-derived cells and resident cardiac cells.