bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 505: Effects of Etamicastat, a Selective Peripheral Dopamine-β-Hydroxylase Inhibitor, Upon Stress-induced Changes in Blood Pressure in the SHR
Authors: Igreja Bruno , Pires Nuno , Wright Lyndon C , Soares-da-Silva Patricio
Published: March 2022

Stressors induce an overactivation of the sympathetic nervous system (SNS) that triggers several changes in the body as a response to stress. These changes include vasoconstriction and increase in heart rate, which results in raised blood pressure (BP). Etamicastat is a selective dopamine-β-hydroxylase (DβH) inhibitor that decreases norepinephrine levels in peripheral sympathetically innervated tissues.

Cardiovascular parameters were measured after a single oral administration of etamicastat (30 mg/kg) 9 h post-administration in male spontaneously hypertensive rats (SHR) by radio-telemetry or tail-cuff recording. Telemetered recording was performed in both restrained and unrestrained animals.

Etamicastat treatment had no effect on heart rate (HR, beats/min) in any recording method. HR measured in unrestrained SHR instrumented with radio-telemetry probes (305±5) consistently yielded HR values significantly lower than restrained animals (362±11) or those recorded by tail-cuff (393±7). On the other hand, single-dose oral etamicastat treatment decrease in mean arterial pressure (MAP, mm Hg) was -14.0±2.0 from 153.3±4.0 in telemetered unrestrained animals, -26.1±5.1 from 182.8±6.4 in telemetry-implanted restrained SHR and -47.9±7.0 from 183.4±9.5 recorded by tail-cuff method. In addition, a power spectral analysis was performed to evaluate BP variability (BPV), in SHR implanted with radio-telemetry probes, at high frequency (HF: 1-2 Hz), low frequency (LF: 0.2-0.6 Hz) and very low frequency (VLF: 0.02-0.2 Hz). The stressor (i.e., cage restrainement) increased the ratio LF/HF-BPV in untreated SHR from 5.23±1.24 to 10.03±1.75, which results from the activation of the SNS. Treatment with etamicastat decreased the ratio LF/HF-BPV in stressed SHR from 10.03±1.75 to 4.46±0.68 and in non-stressed SHR from 5.23±1.24 to 2.23±0.39.

In conclusion, etamicastat treatment reduces (1) high BP in SHR, (2) the stress-induced increase in SNS drive and (3) the stress-induced increase in BP.
Abstract 506: Blood Pressure-lowering Effects Of Anti-hypertensive Drugs And Combinations: Meta-regression Of Published Clinical Trials. The Atom Study
Authors: Paz Marco , De la Sierra Alejandro , Saez Marc , Barceló-Radó Maria Antonia , Rodriguez Juan J , Vera Pilar , Lagarón Cristina , Tarilonte Patricia , Garrido Jose M , Coll de Tuero Gabriel
Published: March 2022

Background and aim:
The use of antihypertensive drugs should be based on the knowledge of their blood pressure (BP)-lowering effect . Most of the clinical trials only assess such effects by comparing 2 different drugs or treatment strategies. However, the relative effect of major antihypertensive drug classes or the most used compounds inside each class is less known. The aim of the study was to assess the BP-lowering effect of most commonly used drugs or drug combinations, by reviewing data from clinical trials based on drug efficacy.

Patients and methods:
Clinical trials with at least 50 patients recruited, in which BP reduction was the primary outcome, were obtained from a systematic review of Medline and Cochrane Central Register of Controlled Trials. Studies without clearly established data of doses or baseline or final BP values were excluded. Data were analyzed by Bayesian meta-regression, adjusted by baseline BP, age, gender, ethnicity, and number of patients included. Only those drugs with a minimum number of studies, which allowed the Bayesian meta-regression, were included.

Results:
253 studies including 937 treatment arms were reviewed. Total number of patients included was 138,971. Systolic BP reduction with monotherapy did not, generally, overcome 20 mmHg, with bisoprolol (mean -15,6 mmHg, 95% CI: -27,4; -1,7) and olmesartan (-15,3 mmHg, 95% CI -18.2; -11,7) resulting the most efficacious drugs. Diastolic BP was reduced by 10 mmHg; being olmesartan the drug promoting a more pronounced reduction (-11.5 mmHg, 95% CI: -13.5; - 9.4). In studies using antihypertensive combinations, the greatest systolic BP reduction was obtained with olmesartan/amlodipine (-31.6 mmHg, CI: -54.6; -22.6) and valsartan/hydrochlorothiazide (-31.4 mmHg, CI: -39.0; -23.0).

Conclusions:
This systematic review reflects the expected BP reductions with the use of the most common antihypertensive compounds or combinations. The results may allow the clinician to choose the most appropriate treatment taking into account patients’ BP and the required reduction to reach the goal.
Abstract 508: Phase 3, Multicenter, Randomized, Double-Blind, Placebo-Controlled Trial of Once-Daily ZS-9 for Treatment of Hyperkalemia: Maintenance of Serum K
+
in Subgroup Analysis of Patients on RAAS Therapy
Authors: Singh Bhupinder , Rasmussen Henrik S , Lavin Philip T , Yang Alex , Roger Simon D
Published: March 2022

Renin-angiotensin-aldosterone system inhibitors (RAASi) increase the risk of hyperkalemia (HK; serum potassium [K
+
] ≥5 mEq/L), often leading to treatment discontinuation, despite proven benefit in chronic kidney disease (CKD) and heart failure patients. There is an unmet need for safe, effective HK treatments which have sustained reduction in K
+
. ZS-9 is a novel, nonabsorbed cation exchanger specifically designed to entrap excess K
+
. Change in serum K
+
in the subgroup of patients receiving RAASi in a Phase 3 trial of ZS-9 are presented.

Patients (N=753) with K
+
5.0-6.5 mEq/L were randomized (1:1:1:1:1) to ZS-9 (1.25, 2.5, 5 or 10 g) or placebo orally 3x daily (TID) for 48 hr (acute phase), following which patients with K
+
3.5-5.0 mEq/L (n=542) were simultaneously randomized 1:1 to the same dose of ZS-9 given during the acute phase or placebo 1x daily (QD) on Days 3-15 (extended phase). Patients who were on placebo during the acute phase were re-randomized to 1.25 or 2.5g ZS-9 QD. RAASi therapy was maintained during the study. Unpaired t-test was used to compare serum K
+
in the RAASi subset that were treated with the two highest doses of ZS-9 (5 or 10g) vs placebo.

A total of 502 of 753 patients (67%) were on RAASi at baseline (ACE inhibitor 66%; angiotensin receptor blocker, 36%; spironolactone 9%). Mean baseline K
+
was 5.3 mEq/L. At 48 hr, significantly greater decreases in K
+
were seen with both ZS-9 dose groups (n=95 each for 5g or 10g; -0.53 and -0.73 mEq/L, respectively) vs placebo (n=98; -0.24 mEq/L; p<0.001). Of the 502 patients on RAASi, 263 entered the extended phase, including 37 patients who continued on 5g QD, 43 who switched to placebo, 43 who continued on 10g QD, and 39 who switched to placebo. During the extended phase, normokalemia was maintained in the10g dose group, whereas K
+
levels increased in patients who switched to placebo (K
+
4.6 vs 5.0 mEq/L, respectively; p<0.001). Similar results were observed in patients who continued on 5g ZS-9 QD, compared with those who switched to placebo (K
+
4.8 vs. 5.1 mEq/L, respectively; p=0.002), at the end of the extended phase.

ZS-9 (5 or 10g) achieved and maintained mean serum K
+
<5.0 mEq/L in patients on RAASi, thus potentially enabling optimal dosing of cardio- and reno-protective RAASi in patients with heart failure or CKD.
Abstract 515: Serotonin Relaxes the Superior Mesenteric Vein: Implications for Blood Pressure Control
Authors: Watts Stephanie W , Darios Emma , Seitz Bridget M , Burnett Robert , Thompson Janice M
Published: March 2022

Serotonin (5-HT, 5-hydroxytryptamine) reduces blood pressure of the conscious rat when administered chronically (one week). 5-HT does not directly relax isolated arteries, and microsphere experiments in 5-HT-infused rats suggested that 5-HT increased flow to the splanchnic bed. We hypothesized that 5-HT increased splanchnic flow because of direct venous relaxation, and not indirect removal of arterial sympathetic tone. Surgical removal of sympathetic innervation of the splanchnic circulation did not alter the ability of 5-HT to lower blood pressure, allowing us to focus on the venous vasculature. Real time RT-PCR, immunohistochemistry and Western analyses supported the predominant expression of the 5-HT2B and 5-HT7 receptor in the superior mesenteric vein. The superior mesenteric vein was mounted in tissue baths for measurement of isometric contraction. 5-HT caused a concentration-dependent relaxation of the endothelin-1 (ET-1) contracted vein. The threshold of 5-HT-induced venous relaxation was significantly lower than for 5-HT-induced venous contraction (1 nM vs 1 μM, respectively). A series of serotonergic agonists was tested, and 5-carboxamidotryptamine (5-CT; mixed 5-HT1, 5-HT7 receptor agonist) and 5-HT were most potent in causing relaxation. Consistent with these findings, the 5-HT7 receptor antagonists, SB 266970 and LY215840, but not the 5-HT2B receptor antagonist LY272015 abolished 5-CT-induced relaxation of the isolated superior mesenteric vein (figure). These studies support the ability of 5-HT to directly relax veins, thereby increasing venous capacitance with a concomitant fall in blood pressure.
Abstract 519: Increased Renal Angiotensinogen Expression in Non-clipped Kidneys of 2-Kidney 1-Clip Hypertensive Rats
Authors: Navar L Gabriel , Shao Weijian , Satou Ryousuke , Prieto Minolfa C , Miyata Kayoko , Katsurada Akemi , Mitchell Kenneth D
Published: March 2022

In 2-kidney 1-clip (2K1C) hypertension, intrarenal angiotensin II (Ang II) levels are increased in both kidneys but the mechanisms for augmentation of Ang II may be different. We recently reported that urinary angiotensinogen (AGT) is increased only in the non-clipped kidneys. However, it has not been determined if these changes are accompanied by augmentation of intrarenal AGT mRNA synthesis in the non-clipped kidneys of 2K1C hypertensive rats. Experiments were performed on male Sprague-Dawley rats (n=11) subjected to left renal arterial clipping (.25 mm gap) and followed for 18-21 days prior to anesthesia and separate measurements of renal function. Systolic arterial pressure increased to 180±3 mmHg compared to 126±4 mmHg in sham operated rats. There were no significant differences in water intake, body weights, and 24 hour urine volume and sodium excretion in awake rats. Separate measurements of renal function showed that renal plasma flow and glomerular filtration rate were similar in clipped and non-clipped kidneys and not different from those in sham rats. However, urine flow, sodium excretion and urinary AGT (uAGT) excretion were significantly greater in non-clipped kidneys compared to clipped and sham kidneys. While kidney AGT protein levels were not increased significantly, AGT transcript measured by real time RT-PCR revealed that the AGT mRNA levels in the cortex were 2.15 fold significantly greater in the non-clipped kidneys than in sham (1.0±.1) or clipped kidneys (.98±.15). The results support the hypothesis that in the non-clipped kidneys of 2K1C rats there is an augmentation of intrarenal AGT mRNA synthesis which explains increased uAGT excretion rates and intrarenal Ang II levels.
Abstract 521: Cardiovascular Phenotype in Mice with Renovascular Hypertension and Type II Diabetes
Authors: Al-Saiegh Yousif , Knudsen Bruce E , Hartono Stella , Grande Joseph P
Published: March 2022

Background:
Combined renovascular hypertension (RVH) and type II diabetes (DM II) are a major health problem. Both share common risk factors and contribute to cardiovascular morbidity and mortality. We recently observed a high mortality rate in DM II mice (dbdb) with RVH.

Purpose:
We sought to determine the interactions between renovascular hypertension and DM II on the cardiovascular phenotype in dbdb mice with RVH.

Methods and Materials:
RVH was established by placing a polytetrafluoroethylene cuff on the right renal artery of C57BL/KS (wild type, WT) or dbdb mice.

We divided the mice into four groups: WT sham, WT type RAS, dbdb sham, dbdb RAS. We examined the blood pressure, plasma angiotensin I, renin, heart weight and myocyte diameter. Histologic parameters including inflammation, necrosis and fibrosis were assessed semiquantitively.

Results:
Blood pressure, Renin, Plasma angiotensin I: RAS produced a similar elevation in systolic blood pressure (SBP), renin mRNA expression, and plasma angiotensin I content in WT and dbdb mice with RAS.

Heart weight:
Despite similar elevation in SBP, db/db RAS mice showed a greater increase in heart weight than WT RAS mice (163.7 +/- 6.4 mmHg vs 145.7 +/- 2.3; n=23).. Similar trends were observed on normalization of the heart weight to tibial length.

Cardiac myocyte diameter:
Cardiac myocyte diameter in WT RAS (n=26) was significantly higher than in WT sham (n=15; 16.4 ± 0.3 vs. 15.0 ± 0.3 μm, p=0.0031). Myocyte diameter was higher in dbdb RAS mice (n=19, 14.76 ± 0.35) than dbdb sham (15.94 ± 0.29 μm, p= 0.0126). However, myocyte diameter was similar in dbdb RAS and WT RAS mice (16.4 ± 0.3 vs. 15.9 ± 0.3 μm, p=0.3).

Histology:
Sham groups had no inflammation (INF) or necrosis (NEC). Interstitial fibrous tissue was increased in dbdb RAS mice (28% by area) vs WT RAS (19% by area). Dbdb RAS mice had more severe INF (severe in 18%, mild in 82%) than WT RAS (severe in 0%, jild in 12%). Severe NEC was found in 32.1% of dbdb compared to 11.5% of WT RAS.

Conclusion:
The histologic changes and hypertrophy of the heart superimpose in dbdb RAS compared to WT RAS. This indicates that the risk of cardiovascular morbidity and mortality increases synergisticly in the presence of DM II and RVH.
Abstract 526: Blood Pressure in the SHR Following Renal Denervation or Etamicastat, a Novel Dopamine-β-Hydroxylase Inhibitor
Authors: Pires Nuno , Igreja Bruno , Moura Eduardo , Wright Lyndon C , Serrão Paula , Soares-da-Silva Patricio
Published: March 2022

Overactivity of the sympathetic nervous system plays an important role in the development and progression of hypertension. Catheter-based renal nerve ablation for treatment of drug-resistant hypertension has been recently developed. An alternative strategy for modulation of sympathetic nerve function is to reduce the biosynthesis of norepinephrine (NE) via inhibition of dopamine β-hydroxylase (DβH), the enzyme that catalysis the conversion of dopamine to NE in sympathetic nerves.

Renal denervation (RDN) was performed in spontaneously hypertensive rats (SHR) to evaluate the effect of RDN on NE levels and blood pressure throughout a 28-day period. Chronic oral treatment with the peripheral selective DβH inhibitor etamicastat (30 mg/kg/day) was performed in another cohort of SHR.

RDN and chronic oral treatment with etamicastat did not affect renal function, as assessed by urinary protein, creatinine and urea levels. RDN significantly decreased mean arterial blood pressure (MAP, in mm Hg) 7 days post-surgery (139±6, P<0.01) versus baseline values (158±6). A gradual MAP return to high baseline levels was observed overtime; 139±7 at day 14, 151±12, at day 21 and 155±12 at day 28 after RDN. Treatment with etamicastat decreased baseline MAP (150±5) in a sustained and significant (P<0.01) manner at all-time points: 141±5 at day 7, 139±5 at day 14, 138±5 at day 21 and 143±5 at day 28. At day 28, NE levels (in pmol/mg tissue) in renal tissue were significantly decreased in both RDN (0.12±0.08, P=0.001) and etamicastat-treated group (0.48±0.04, P=0.01) as compared to vehicle group (0.88±0.13). On the other hand, left ventricle NE levels were decreased only after treatment with etamicastat (1.24±0.12, P=0.04) but not in the RDN group (2.53±0.24, P=0.9) when compared to controls (2.59±0.58).

It is concluded that RDN produces transitory decreases in MAP, whereas prolonged down regulation of sympathetic drive with the DβH inhibitor etamicastat results in sustained decreases in MAP.
Abstract 530: Enhanced Expression Of Epithelial Sodium Channels (ENaC) Causes Salt-induced Hypertension In Mice Through Inhibition Of The α
2
-isoform Of Na
+
, K
+
-atpase
Authors: Leenen Frans , Hou Xiaohong , Wang Hongwei
Published: March 2022

Knockout of the Nedd4-2 gene in mice results in overexpression of ENaC on the plasma membrane in kidneys, choroid plexus and brain nuclei. These mice exhibit enhanced pressor responses to CSF [Na
+
] and dietary salt-induced hypertension which both can be blocked by central infusion of the ENaC blocker benzamil. Functional studies suggest that ENaC activation in the CNS results in release of endogenous ouabain (EO) and inhibition of the α
2
-isoform of Na
+
, K
+
-ATPase. To test this concept more specifically, we studied Nedd4-2
-/-
mice expressing the α
2
R/R
-isoform of Na
+
, K
+
-ATPase. Intracerebroventricular (icv) infusion of Na
+
-rich aCSF (225 mM Na
+
at 0.4 μl/min) increased MAP by 10-15 mmHg in wild type mice and by 25-30 mmHg in Nedd4-2
-/-
mice, but by only ~5 mmHg in α
2
R/R
and in α
2
R/R
/Nedd4-2
-/-
mice (Fig top panel). Icv infusion of EO-binding Fab fragments also blocked the BP response in Nedd4-2
-/-
mice. In Nedd4-2
-/-
mice high salt diet increased MAP by 25-30 mmHg, but in α
2
R/R
/Nedd4-2
-/-
mice by only 5 mmHg (Fig bottom panel). These findings substantiate the concept that enhanced ENaC activity causes salt-induced pressor responses mainly through EO inhibiting the α
2
-isoform of Na
+
, K
+
-ATPase in the brain.
Abstract 532: GRK4 Palmitoylation is Necessary for Membrane Association and Dopamine-1 Receptor Activity
Authors: Gildea John J , Huang Zhenyue , Bigler Wang Dora , Tran Hanh , Felder Robin A
Published: March 2022

The renal dopaminergic system regulates a large percentage of renal sodium excretion under sodium loaded conditions; however, not much is known about intracellular dopaminergic regulatory mechanisms. The expression of the principal sodium regulatory dopaminergic receptor (the D1R) is negatively modulated by the G protein linked receptor kinase type 4 (GRK4) through phosphorylation while the D1R is expressed in the plasma membrane. Therefore, we hypothesized that mechanisms associated with GRK4 membrane localization might be associated with its activity. Using human renal proximal tubule cell lines (RPTC), we previously demonstrated that GRK4 is palmitoylated using 17-octadecynoic acid (17-ODYA) and click chemistry and that fenoldopam (FEN, D1R agonist) decreased palmitoylation only in normal cAMP-coupled RPTC. Here we expand on those studies by showing that GRK4 palmitoylation was inhibited by 2-bromopalmitate (2BP, 100 μmol/L, palmitoylation inhibitor) which shifted GRK4 expression from the membrane to the cytoplasm. We used two different techniques to show loss of GRK4 membrane association when GRK4 palmitoylation was inhibited. The first technique involved classic cell fractionation using ultracentrifugation and western blot identification of GRK4 in membrane protein fractions (VEH 0.174±0.023 RFU, 2BP 0.091±0.013 RFU, P<0.05, N=3) The second technique utilized a novel membrane-associated fluorescent linked immunosorbent assay (mFLISA) to demonstrate the decrease in GRK4 membrane association (VEH 13,568±664 RFU, 2BP 5,155±974 RFU, P<0.05, N=12). Additionally, we showed that inhibiting GRK4 palmitoylation ledto D1-like plasma membrane recruitment using a non-cell permeable fluorescent D1-like antagonist bodipy630 SKF83566 (VEH 58,242±3858 RFU, 2BP 80,153±4061 RFU, P<0.01, N=6). In summary, we demonstrated a novel D1R regulatory mechanism through GRK4 palmitoylation, affecting its localization and activity.
Abstract 534: Cytochrome P450 epoxygenase regulates Na transport in aldosterone-sensitive distal nephron: a lesson learns from Cyp2c44 knockout mice
Authors: Wang Wen-Hui , Imig John , Capdevila Jorge
Published: March 2022

Previous study demonstrated that the down-regulation of epoxygenase caused salt-sensitive hypertension. We used the real-time PCR technique to demonstrate that a high Na (4% Na) intake increased the Cyp2c44 expression in the thick ascending limb and distal convoluted tubule (DCT) and connecting tubule (CNT). The disruption of cyp2c44 increased ENaC activity in CNT and the 40 pS inwardly-rectifying K channel, a heterotetramer of Kir.4.1/Kir5.1, in the basolateral membrane of early segment of DCT (DCT1). Application of epoxyeicosatrienoic acid (EET) inhibited both ENaC in the CNT and the Kir4.1/5.1 channels in DCT1. Kir4.1/5.1 plays a key role in generating the cell membrane potential in the DCT and is the only type of K channels responsible for providing K conductance in the basolateral membrane of the DCT. Also, we demonstrated that Kir.4.1/5.1 activity determines the apical NCC expression and Ste20-related-proline-alanine-rich kinase (SPAK) which stimulates NCC activity through the phosphorylation of NCC. Since EET inhibited Kir4/1/5.1 in the DCT, the disruption of Cyp2c44 significantly increased the basolateral Kir4.1/5.1 activity thereby increasing the expression of SPAK and NCC in
cyp2c44
knockout mice. Thus, feeding
Kcnj10
knockout mice with high Na caused hypertension. Moreover, the application of amiloride failed to completely reverse the Na-sensitive hypertension in the knockout mice while the application of thiazide and amiloride restored the normal blood pressure in
Kcnj10
knockout mice. We conclude that Cyp2c44 plays an important role in suppressing NCC in the DCT and ENaC in the CNT during increasing dietary Na intake. The upregulation of NCC expression in
Kcnj10
-/- mice was due to the stimulation of the basolateral K channels in the DCT.
Abstract 539: Remodeling Of Afferent Arterioles From Mice With Oxidative Stress Does Not Enhance Contractility But Does Limit Excessive Wall Stress
Authors: Li Lingli , Lai En Yin , Welch William J , Wilcox Christopher S
Published: March 2022

Background:
Remodeling of resistant vessels in hypertension is postulated to enhance their contractility. We showed that knockout of SOD increased superoxide (O
2
.-
), contractility and remodeling of afferent arterioles (Affs). Therefore, we tested the hypothesis that the remodeling causes the enhanced contractility.

Methods:
Myogenic responses (MRs) were assessed from the slope of the regression of active wall tension (AWT) on perfusion pressure (PP) in wild type and knockout SOD-1,-2 or -3 mice and in mice 3 days after induction of SOD-3 knockout before remodeling had developed. Contractions to 10
-6
M Ang II were tested also. O
2
.-
was assessed from PEG-SOD inhibitable ethidium:dihydroethidium fluorescence and remodeling from media:lumen area ratio (M:L).

Results:
1. Compared to +/+, increasing PP or 10
-6
M Ang II increased O
2
.-
of Affs more in SOD1-/-, SOD2+/- and SOD3-/- (P<0.01). 2. Compared to +/+, MRs (dynes.cm
-1
.mmHg
-1
) were significantly (P<0.05) increased in SOD1-/- (3.49 ± 0.30 vs 2.30 ± 0.27), in SOD2+/- (3.85 ± 0.48 vs 1.80 ± 0.34), and in SOD3-/- (3.51 ± 0.20 vs 2.43 ± 0.15). 3. Compared to +/+, Ang II contractions were also stronger in knockout mice (P<0.001). 4. Compared to +/+, M:L ratio of Affs were significantly (P<0.001) greater in SOD1-/- (4.0 ± 0.2 vs 1.8 ± 0.2), in SOD2+/- (3.7 ± 0.4 vs 1.8 ± 0.4), and in SOD3-/- (4.5 ± 0.2 vs 1.9 ± 0.1). 5. Across genotypes, MR (r
2
=0.32, P<0.005) and Ang II contractions (r
2
= 0.56, P<0.005) were strongly correlated with M:L ratios. 6. Changes in AWT (T
physiol
- T
passive
) during increase in PP were dependent on SOD genotype (P<0.01) yet changes in vessel wall stress (AWT/wall thickness) were independent (P=NS). 7. Compared to controls , MRs and Ang II contractions were increased 3 days after inducing SOD3 knockout (MR, 3.76 ± 0.55 vs 2.26 ± 0.01, P<0.05; Ang II, -66 ± 4% vs -49 ± 4%, P<0.01) and were not significantly different from SOD3-/- mice. However, the M:L ratio of these Affs was unchanged (2.0 ± 0.1 vs 1.8 ± 0.2, NS).

Conclusions:
lifetime increases in cytosolic, mitochondrial or extracellular O
2
.-
enhances afferent arteriolar contractility and remodeling. The remodeling is not required for the enhanced contractility, but does prevent the damaging effects of increased wall stress.
Abstract 548: Anti-aging Gene Klotho Deficiency Exacerbates High Fat Diet-induced Arterial Stiffening and Hypertension by Downregulation of AMP-activated Protein Kinase
Authors: Lin Yi , Velury Divya , Sun Zhongjie
Published: March 2022

Background:
Arterial stiffening and hypertension are progressive aging-related disorders. Klotho is an anti-aging protein but its role in regulation of arterial stiffening and hypertension is poorly understood.

Methods and Results:
Heterozygous
Klotho
deficiency (
KL
+/-
) mice and WT littermate mice were fed on high fat diet (HFD) or normal diet (ND). Pulse wave velocity (PWV), an index of arterial stiffening, was increased in
KL
+/-
mice fed on HFD for 5 weeks. Systolic blood pressure and blood glucose levels increased earlier with greater magnitudes in
KL
+/-
mice fed on HFD. Notably, protein expression of collagen I, Runx2, and TGFβ1 were increased but protein expression of phosphorylated AMPKα (pAMPKα), phosphorylated eNOS (peNOS), and Mn-SOD were decreased in aortas of
KL
+/-
mice fed on HFD for 5 weeks. Interestingly, daily injection of AICAR, an activator of AMPK, abolished the increases in PWV, blood pressure, and blood glucose in
KL
+/-
mice fed on HFD. AICAR not only abolished the downregulation of pAMPKα, peNOS, and Mn-SOD levels but also attenuated the increased levels of collagen I, Runx2, TGFβ1 and superoxide, elastic lamellae breaks, and calcification in aortas in
KL
+/-
mice fed on HFD.

Conclusions:
Klohto deficiency promotes HFD-induced arterial stiffening and hypertension. The promoting effects of klotho deficiency on arterial stiffening may be due to downregulation of AMPKα activity.
Abstract 550: Plasma Angiotensin-(1-12) Level Is Elevated And Correlates With Proteinuria In Patients With Chronic Kidney Diseases
Authors: Moniwa Norihito , Furuhashi Masato , Yoshida Hideaki , Takizawa Hideki , Hasegawa Koichi , Shimamura Yoshinosuke , Mita Tomohiro , Fuseya Takahiro , Ishimura Shutaro , Tanaka Marenao , Ura Nobuyuki , Tanaka Shigemichi , Miura Tetsuji
Published: March 2022

Angiotensin-(1-12) [Ang-(1-12)], C-terminally extended form of Ang I, is an alternative precursor of Ang II. Although angiotensinogen, a precursor of both Ang I and Ang-(1-12), is reportedly one of markers of kidney disease, little is known about role of Ang-(1-12) in pathological status especially in human. To address this issue, we measured plasma and urinary Ang-(1-12) by enzyme immunoassay in 4 healthy volunteers and 15 patients with biopsy-proven chronic kidney diseases (CKD). CKD group included minor glomerular abnormality (n=3), IgA nephropathy (n=8), minimal change nephritic syndrome (n=4). Ang-(1-12) level was 55-fold higher in urine than in plasma (Figure; 4.12±1.55 vs 0.075±0.006 ng/mL, P<0.05). Neither plasma Ang-(1-12) nor urinary Ang-(1-12) was correlated with blood pressure. Plasma Ang-(1-12) was positively correlated with urinary protein and tended to be negatively correlated with estimated glomerular filtration ratio (eGFR; Figure). Plasma Ang-(1-12) was higher in the CKD group than in the control (Figure; 0.083±0.006 vs 0.045±0.005 ng/mL, P<0.01). On the other hand, urinary Ang-(1-12) was correlated with neither urinary protein nor eGFR. Urinary Ang-(1-12) tended to be higher in the CKD group than in control (4.94±1.9 vs 1.05±0.6 ng/mL, respectively). The present study showed for the first time that plasma Ang-(1-12) level is elevated in patients with CKD and suggests that increased plasma Ang-(1-12) plays a nephropathic role.
Abstract 551: Melatonin Prevents Renal Injury in a High Salt Diet Induced Hypertension Model by Decreasing Oxidative Stress.
Authors: Leibowitz Avshalom , Volkov Alexander , Barshack Iris , Grossman Ehud
Published: March 2022

Oxidative stress plays an important role in the pathogenesis of target organ damage (TOD) in hypertension, including renal injury. There is evidence, from human and animals' models, that melatonin is involved in the regulation of blood pressure (BP) and may protect from development of high BP and TOD . Melatonin is one of the potent antioxidant molecules in nature. We hypothesized that melatonin protective effect in a high salt diet (HSD) rodent model is mediated by decreasing oxidative stress. Ten Dahl salt sensitive (DSS) rats were fed normal chow [control (Ctrl)], 10 were fed HSD and 10 were fed HSD and got melatonin (Mel) (30/mg/kg/day) in their water for 8 weeks. Body and kidneys weights were measured at the end of the treatment. Renal function was evaluated by measurement of 24 hours urine protein excretion (n=8-10). Glomerular injury index (GII) (fibrotic glomerulus/ 100 glomerulus) was evaluated in Masson's trichrom stained section of renal cortex (n=5). Oxidative stress in the kidneys was evaluated by determining superoxide production using dihydroethidium (DHE) staining (n=5). Melatonin prevents weight loss caused by HSD (Ctrl-398±8, HSD- 335±17 Mel- 372±7 gr. P<0.05 HSD vs. Ctrl and Mel) but not the renal hypertrophy (Ctrl-0.58±0.02, HSD- 0.87±0.04 Mel- 0.9±0.03 gr. /tibia length, p<0.05 Ctrl vs. HSD and Mel). Melatonin decreased the proteinuria seen in HSD fed rats (HSD- 50.7±12 Mel- 22.3±4.3 Protein/Cr, p<0.05). Glomerular damage was significantly higher in the HSD group (GII 24±6) than in Ctrl group (GII 0.8±0.5, p<0.05). Melatonin diminished significantly this damage (GII 3.6±0.8, p<0.05). Superoxide production was significantly higher in kidneys of HSD fed rats than in the Ctrl rats (99±9 vs. 60±7 relative fluorescent units (RFU)/μm
2
, respectively, p<0.05).Treatment with melatonin decreased superoxide production in the kidneys of HSD fed rats(74±5 RFU/ μm
2
, p<0.05 vs. Ctrl and HSD). Treatment with melatonin abolishes the deleterious effect of HSD on the kidneys of HSD fed rats. Melatonin beneficial effect may be mediated by its anti-oxidative properties.
Abstract 558: The eight-item Morisky Medication Adherence Scale (MMAS-8) score was associated with glycaemic control in diabetes patients
Authors: Wu Carmen H , Wong Martin C , Wang Harry H , Kwan Mandy W , Chan Wai Man , Li Heung Wing , Hui Eric M , Lam Augustine T
Published: March 2022

Background:
Adherence with oral hypoglycaemic agent is important to achieve optimal glycaemic control. The eight-item Morisky Medication Adherence Scale (MMAS-8) is frequently used for measuring drug adherence, yet the association between MMAS-8 score and glycaemic control among diabetes patients has not been addressed.

Methods:
Two general outpatient clinics were randomly selected in a district with socio-demographic representative of the entire Hong Kong population. In 2013, all type II adult diabetes patients on oral hypoglycaemic agents were consecutively invited to participate in a self-administered survey with the validated Chinese version of MMAS-8. The glycaemic control was reflected by HbA1c taken in the clinic in the same occasion. Factors associated with poor glycaemic control (HbA1c ≥ 7.0%) were explored by a binary logistic regression analysis.

Results:
From 565 eligible Chinese patients with an average age of 63.2 years (SD 9.7) and male proportion of 46.5%, the average HbA1c was 7.14% (SD 1.1%), and 48.0% [271/565] had poor glycaemic control (HbA1c ≥ 7.0%). The proportion of poor medication adherence (MMAS-8≤ 6) was 32.2% [182/565]. After controlling for age, sex, education level, BMI, income, smoking, alcohol drinking, compliance with diabetes diet, and use of chronic medications (antihypertensive agents; lipid-lowering agents; antiplatelets), the higher MMAS-8 score (better drug adherence) was significantly associated with HbA1c level (Beta -0.115; 95% C.I. -0.164 to -0.026, p = 0.007).

Conclusion:
The MMAS-8 score was negatively correlated with HbA1c, further strengthening its validity in prediction of glycaemic control among diabetes patients.
Abstract 572: Transcriptional Regulation Of Renal Dopamine D1 Receptor Function During Oxidative Stress
Authors: BANDAY ANEES A , Lokhandwala Mustafa F
Published: March 2022

There exists a strong link between Oxidative stress, renal dopaminergic system and hypertension. It is reported that reactive oxygen species attenuate renal proximal tubular dopamine receptor (D1R) function which disrupts sodium regulation and leads to hypertension. The mechanisms for renal D1R dysfunction, however are not clear. Here in, we investigated the role of redox sensitive transcription factors AP1 and SP3 in transcriptional suppression of D1R gene function and subsequent D1R signaling. Human kidney proximal tubular (
HK2
) cell were treated with a pro-oxidant L-buthionine sulfoximine (
BSO
, 50 mM for 72 hr) with and without a polyphenol resveratrol (250 μM). Treatment of HK2 cells with BSO caused oxidative stress (malondialdehyde [MDA], nmol/mg protein: control: 0.13 ± 0.01, BSO: 0.20 ± .02) and significantly reduced D1R mRNA and membrane receptor number (fmol/mg protein; control—140.1 ± 5.6, BSO—81.7 ± 4.4). Incubation of HK2 cells with SKF38393, a D1R agonist, caused a concentration dependent inhibition of Na/K-ATPase (
86
Rb uptake, nmol/mg protein, control: basal—9.2 ± 0.2, SKF (1 μM)—6.3 ± 0.1). However, SKF38393 failed to inhibit Na/K-ATPase in BSO-treated cells (BSO: basal—9.5 ± 0.3, SKF—8.6 ± 0.2). Cells incubated with BSO exhibited increased AP1 (c-fos) and SP3 nuclear expression. Resveratrol per se had no effect on oxidative milieu (MDA: 0.11 ± 0.01) or D1R expression (143.3 ± 4.4), but it mitigated BSO-induced oxidative stress (MDA: 0.13 ± 0.02), AP1—SP3 up-regulation and D1R down-regulation (133.6 ± 6.2) and dysfunction (resveratrol + BSO [
86
Rb uptake]: basal—9.7 ±0.3, SKF—6.7 ± 0.4). Incubation of cells with AP1 or SP3 specific siRNA reduced the expression of respective protein by ~85% and abolished BSO-induced D1R dysfunction. Down-regulation of SP3 had no effect on BSO-mediated AP1 activation, however, AP1 siRNA blocked BSO-dependent SP3 up-regulation. In cells transfected with Nrf2 siRNA, resveratrol failed to block BSO-induced AP1—SP3 activation and D1R dysfunction. These data shows that oxidative stress

via AP1—SP3 activation suppresses D1R transcription which leads to D1R dysfunction. Resveratrol activates Nrf2, mitigates oxidative stress, blocks AP1—SP3 signaling and prevents D1R down-regulation and dysfunction.
Abstract 600: Aldosterone-Dependent Transfer of microRNA-Containing Exosomes Between Endothelial and Smooth Muscles Cells.
Authors: Robertson Stacy , Dababneh Edward , Bursill Christina
Published: March 2022

Background:
Aldosterone is a cardiovascular risk factor and recent research has demonstrated that aldosterone increases exocytosis from endothelial cells, which may be an important signalling mechanism. Exosomes are small extracellular particles that contribute to cell-to-cell communication. Once released from a host cell they can circulate in the blood and transfer their contents (e.g. miRNAs) to recipient cells. We aimed to examine the effect of aldosterone on: 1) exosome release from endothelial cells and 2) exosome trafficking of miRNA from endothelial cells to smooth muscle cells.

Methods and Results:
Human coronary artery endothelial cells (HCAECs) were incubated with aldosterone (100nM) or vehicle for 1 or 18 hours. Exosomes were isolated from cell media via ultracentrifugation and then analyzed by nanoparticle-tracking (NanoSight). Aldosterone increased exosome concentration by 2.65-fold (1h; 19.1x10
10
particles/mL) or 3.25-fold (18h; 28.6x10
10
particles/mL) when compared to the time-point vehicle control (7.2x10
10
and 8.8x10
10
particles/mL, respectively). To test exosome paracrine trafficking, HCAECs were transfected with a
c.elegans
miRNA (cel-miR-39) and incubated adjacent to human coronary artery smooth muscle cells (HCASMCs) separated by a 0.4μm filter for 24 hour, with or without aldosterone. qRT-PCR analysis was used to measure cel-miR-39 expression in HCASMCs following incubation near transfected HCAECs, a confirmation of trafficking. Importantly, levels of cel-miR-39 in HCASMCs increased significantly (2.42±0.52 fold; p=0.02) in aldosterone-stimulated cells, compared to control. Blocking transcription with Dactinomycin had no effect on aldosterone-mediated trafficking (cel-miR-39 expression 2.07±0.61 fold; p>0.05). Finally, antagonizing the mineralocorticoid receptor (MR) with spironolactone had no effect on cel-miR-39 expression in HCASMCs (3.43±1.49 fold; p>0.05).

Conclusion:
This study demonstrates that aldosterone increases exosome secretion from endothelial cells, leading to increased uptake of exosome-packaged microRNAs by smooth muscle cells, independent of transcription and the MR. Together, this represents a novel mechanism by which aldosterone contributes to vascular disease.
Abstract 602: RAS-induced Hypertension Is Primarily Mediated By (Pro)renin Receptor Activation At The Paraventricular Nucleus Of The Hypothalamus.
Authors: Rodríguez Vermalí , de Kloet Annette D , Chan Tiffany S , Sumners Colin
Published: March 2022

Studies have shown that renin-angiotensin-system (RAS)-induced inflammation at the paraventricular nucleus of the hypothalamus (PVN) contributes to hypertension development by increasing sympathetic outflow and elevating blood pressure. Recent data suggest that PRR activation increases pro-inflammatory cytokine (PIC) production in an Ang II-independent manner helping sustain kidney inflammation, retinal disease and diabetic neuropathy. Astrocytes house all the components of the RAS and help regulate the immune response within the brain. However, the role of (pro)renin (PRO) receptor (PRR) activation at the PVN on blood pressure regulation and astrocyte activation has not been evaluated. We show that at the PVN, 12 weeks old spontaneously hypertensive rats (SHRs) have significantly (p < 0.05) higher PRR mRNA levels than normotensive Wistar Kyoto (WKY) and Sprague Dawley (SD) rats (1.6 ± 0.3 vs. 0.96 ± 0.4 & 1.0 ± 0.2, SHR, WKY and SD respectively). In situ-hybridization data using RNA Scope, confirmed these results as qualitative analysis from coronal sections containing the PVN from SD and WKY rats showed lower PRR signal when compared to those from SHRs. Our data shows that bilateral downregulation of the PRR, using a short-hairpin RNA (shRNA) vector construct, at the PVN of SHRs (n = 5) reduces blood pressure compared to control animals (n = 5) with PRR levels intact (146.1 ± 6.6 vs. 133.3 ± 9.0 mmHG; p < 0.05). Similarly, PRR downregulation at the PVN can also reduce the pressor response triggered by sub-cutaneous Ang II infusion (200ng/kg/min) in SD rats (143 ± 13.2 vs. 114.8 ± 13.2 mmHG, p < 0.05; control vs. shRNA animals, respectively). In both experimental designs, blood pressure decreases were not accompanied by changes in heart rate. Our results also show lower glial fibrillary acidic protein (GFAP) and interleukin (IL)-1β mRNA levels at the PVN in shRNA-injected animals and using the Image J tool we show a trend for astroglial area fraction in the PVN to decrease. These changes are in concert with our in vitro data which show that PRO increases PICs levels and astrogliosis in cultured astrocytes. Our findings show that at the PVN, PRR activation is key in RAS-induced hypertension and potential mechanisms may include astrocyte- and neuroimmune activation.
Abstract 603: Clinic Blood Pressure Prediction of Normal Ambulatory Blood Pressure in Youth
Authors: Johnson Philip K , Ferguson Michael A , Zachariah Justin P
Published: March 2022

INTRODUCTION:
As obesity in youth has increased over time, elevated blood pressure (BP) has also risen. Ambulatory Blood Pressure Monitors (ABPM) may be useful since younger patients have low pretest probability of hypertension and are susceptible to both white-coat and masked hypertension. To aid in appropriate ABPM use within a referral cohort, we sought to define systolic clinic BP (CBP) that predicted normal ABPM and target end organ damage (TOD).

METHODS:
Data were collected on consecutive patients referred for high BP undergoing ABPM from 2010-13 (n=253, 33.3% female, mean age 15.9 ± 3.7 years). Logistic regression models adjusted for age, sex, and height were used to determine the relation between normal ABPM as the outcome and CBP indices, which were sequentially replaced as predictor. CBP indices were defined as systolic maximum, minimum or average auscultative, automated or combined. Relations between TOD and CBP or ABPM were assessed using logistic regression. Receiver operator curves tested the predictive utility of significant CBP indices.

RESULTS:
Lower in-clinic auscultative CBP predicted Normal BP [Table1] and had a c-statistic of 0.61. Auscultative CBP at the 95% threshold had sensitivity of 71%, specificity of 47%, positive predictive value of 58%, and negative predictive value of 61% for normal ABPM. Auscultative maximum CBP did not predict elevated left ventricular mass:volume ratio (OR 1.03 [0.98-1.09]; p=0.23) while ABPM hypertension did (3.44 [1.28-9.24]; p=0.01)

CONCLUSION:
In youth, auscultative CBP was a weak predictor of normal ABPM and did not predict end organ damage. The role of ABPM in general and special populations deserves exploration.
Abstract 613: Mitochondrial Hydrogen Peroxide In T Cell Activation In Hypertension
Authors: Nazarewicz Rafal R , Saleh Mohamed A , Wu Jing , Dikalov Sergey , Harrison David G
Published: March 2022

Substantial evidence demonstrates that mitochondrial reactive oxygen species and adoptive immunity are critical in the development of hypertension. Our initial observations indicated that T cells isolated from hypertensive mice produced more cellular and mitochondrial hydrogen peroxide (H2O2) compared to those from normotensive animals, 98% and 48% respectively. We hypothesized that mitochondrial H2O2 plays a regulatory role in T cell activation in hypertension. We used RAG1-/- male mice that lack lymphocytes and modulated mitochondrial H2O2 by adoptive transfer of T cells with various mitochondrial H2O2 levels. RAG1-/- mice received T cells from wild type mice (RAG1-/-+WT) or mice expressing catalase in mitochondria (RAG1-/-+mCat). Adoptive transfer of T cells was performed 3 weeks prior to Ang II infusion (490 ng/kg/min). Our data show that inhibition of mitochondrial H2O2 specifically in T cells prevents hypertension. When challenged with angiotensin II, systolic BP in RAG1-/-+WT mice rose to 174 mmHg (+/-7.7) which was similar to values observed in WT mice (173 mmHg +/-12). However, scavenging mitochondrial H2O2 in T cells significantly (P<0.01) blunted the hypertensive response in RAG1-/-+mCat mice (135 mmHg +/-9.7). Interestingly, hypertension led to accumulation of CD45+/CD3+ T cell in aorta and kidney of WT mice and RAG1-/- mice that received WT T cells but not those receiving T cells expressing mitochondria catalase. Next, we investigated the mitochondrial metabolism in T cells isolated from hypertensive mice and found that hypertension reduced T cell respiration by 67% and increased ATP production by 54%. These changes were reversed by scavenging mitochondrial H2O2. Our data also show that the rates of glutamine intake and lactate production in T cells are dependent on mitochondrial oxygen species. We propose that mitochondrial hydrogen peroxide plays an important role in regulating T cell metabolism and functions in hypertension.
Abstract 616: Aging, Menopause And Metabolic Syndrome As Related To The Beneficial Effects Of Exercise Training
Authors: Machi Jacqueline F , Bernardes Nathalia , Dias Danielle , Cruz Paula , Moraes-Silva Ivana , Scapini Katia , Nascimento Ademir , Morris Mariana , DE Angelis Katia , Irigoyen Maria Cláudia C
Published: March 2022

Menopause and aging are associated with increased blood pressure. High fructose may modify the development of the pathologies associated with these conditions. The objective was to investigate the effects of exercise training (ET) in aged, oophorectomy mimic of menopause fructose-fed rats. Female Wistar rats (24 months old) were ovariectomized, fed with fructose (100g/L) in drinking water and divided in 5 groups (n=8/group): control (C), ovariectomized (O), ovariectomized trained (OT, n= 8), ovariectomized fructose (OF, n= 8) and ovariectomized fructose trained (OFT, n= 8). The ET was performed on a treadmill (1 h/day; 5 days/wk for 8 wk). The metabolic results showed that ET decreased adipose tissue in both trained groups (OT: 4.3±0.5, OFT: 5.3±0.7), compared with sedentary rats (C: 6.2± 0.6, O: 9.6±1.3, OF: 10.1±0.6, g). The same findings were observed for insulin resistance (OT: 4.6±0.2, OFT: 5.5±0.6 vs C: 3.2± 0.6, O: 3.2±0.3, OF: 4.0±0.5, mg/dl/min). ET increased physical capacity (OT: 20±0.9, OFT: 17.5±0.9 vs C: 11.5± 0.5, O: 10±0.5, OF: 10.3±0.5 Min/h). The hemodynamic results showed that ET attenuated the increase in MBP (C: 109.4± 1.9, OT: 103±1.1, OFT: 107±1.1 vs O: 119.1±1.9, OF: 119.1±2.7, mmHg) and HR (C: 341.4± 8, OT: 302±1, OFT: 306.4±8 vs O: 398.17±19, OF: 378.9±17 Bpm). The sympathetic tonus was decreased in C, OT and OTF (C: 63.6± 4, OT: 62.2±3, OFT: 51.2± 7 beats/ min) when compared with O and OF (O: 102.6±12, OF: 91.9±7 beats/min) while vagal tonus was increased just in OT (OT: 44.8±5) vs (C: 24.9± 6, OFT: 22±3, O: 13±6, OF: 9±2 beats/min). The OT and OTF groups showed an increased bradycardic response to BP changes (C: -1.50±0.06 OT: -1.74±0.12, OFT: -1.77±0.15 vs O: -0.93±0.07, OF: -1.21±0.12 bpm/mmHg). ET increased the Standard deviation of the pulse interval (C: 6.7±1.1 OT: 11.6±0.8, OFT: 13.0±1.5 vs O: 7.05±0.6, OF: 9.57±0.7 ms) and decreased VARR PAS (C: 6.7±1.09 OT: 23.6±0.8, OFT: 30.7±2.6 vs O: 56.4±8.1, OF: 40.5±3.7 mmHg2) as well as representative sympathetic modulation (LF PAS) (C: 6.7±1.1 OT: 3.6±0.5, OFT: 3.7±0.4 vs O: 9.8±1.2, OF: 8.3±0.8 mmHg. In conclusion, ET effectively improved metabolic and cardiovascular parameters altered by aged or excessive fructose consumption in rats submitted to ovarian hormone deprivation.
Abstract 623: Upregulation of Renin-Angiotensin System and Inflammatory Mechanisms in the Prenatally Programmed Hypertension
Authors: Watanabe Ingrid K , Volpini Rildo A , Jara Zaira P , Jung Flavia F , Casarini Dulce E
Published: March 2022

Strong evidence shows that renin-angiotensin system (RAS) is affected by adverse maternal nutrition during pregnancy. Early treatment of offspring with RAS inhibitors is able to prevent the development of prenatally programmed hypertension. The aim of this study was to evaluate renal RAS activity and inflammatory mechanisms and the effects of RAS inhibition in prenatally programmed hypertension. Sprague-Dawley rats were fed with standard diet (control group) or low protein (LP) diet during the period of pregnancy. After birth, dams were switched back to standard rat chow. After weaning, LP pups were randomized to either no treatment or to treatment with losartan. LP offspring presented lower birth weight when compared to CT group (5.3 ± 0.1 vs 6.4 ± 0.2 g, respectively, p<0.05). At 20 weeks of life, blood pressure levels were higher in the untreated LP group when compared to CT group (128.5 ± 0.9 vs 109.2 ± 1.9 mmHg, respectively, p<0.05). We also observed adverse histological features in the kidney of untreated LP animals when compared to CT group, including augmentation of relative interstitial area (9.27 ± 0.32 vs 3.86 ± 0.27 %, respectively, p<0.05), collagen concentration (3.53 ± 0.87 vs 0.68 ± 0.19%, respectively, p<0.05) and number of PCNA positive cells (7.36 ± 1.82 vs 1.95 ± 0.15 cells/0.274 mm2, respectively, p<0.05). Untreated LP group also had higher number of CD3 and CD68 positive cells in the kidney compared to CT group [CD3: 9.3 ± 0.5 vs 5.1 ± 0.3 cells/0.274 mm2 and CD68: 13.5 ± 1.0 vs 8.2 ± 0.9 cells/0.274 mm2, respectively, p<0.05), and higher levels of reactive oxygen species (3.6 ± 0.4 vs 1.7 ± 0.2 mmol/mg, respectively, p<0.05). In addition, renal levels of angiotensin II (Ang II) were higher in the untreated LP offspring compared to CT group (p<0.05), and inhibition of RAS was able to prevent renal injury and abolished infiltration of inflammatory cells, besides its hemodynamics benefits. The finding of high renal levels of Ang II associated with increased inflammatory infiltration and deleterious histological outcomes and the effectiveness of RAS inhibition in prevent them indicate that RAS, via Ang II/AT1 receptor, is responsible for initiating and perpetuating the inflammatory and fibrotic processes in the prenatally programmed kidney.