bluuebunny/crossref_metadata_2025 huggingface dataset:
PS-BPB06-3: EXAMINATION OF THE EFFECTIVENESS OF DIABETIC NEPHROPATHY TREATMENT TARGETING CHREBP ACTIVITY
Authors: Suzuki Susumu , Yokoyama Atsushi , Okamoto Koji , Takahashi Shinichiro , Iwabuchi Yoshiharu , Sugawara Akira
Published: January 2023

Introduction:

Diabetic nephropathy (DN) is the main cause of chronic kidney disease (CKD), and is characterized by elevated urinary albumin to creatinine ratio (UACR) and serum creatinine, and decreased glomerular filtration rate. Anti hypertensive agents are used for DN treatment, however, the effects of these drugs are insufficient, and the establishment of more effective treatment methods is desired.

Carbohydrate responsive element binding protein (ChREBP) is one of the factors reported to be involved in the onset and progression of DN. Furthermore, whole body ablation of ChREBP suggested that the progression of DN was suppressed in the rodent model. On the other hand, the effectiveness of pharmaceutical ChREBP suppression is unclear, thus, we performed administration experiment to DN model mice using ChREBP activity inhibitor and examined the effects of pharmaceutical ChREBP activity suppression on renal function of diabetic mice.

Results:

First, we established high-throughput screening system capable of detecting ChREBP activity and performed screening of Tohoku university compound library. The result compound, D-532, was ChREBP activity specific inhibitor without cell toxicity. D-532 directly binds to DNA-binding domain of ChREBP, and inhibits the transcriptional activity. We next performed administration experiment using type 1 DN model mice. D-532 treatment group showed an improvement tendency of the low body weight compared with that of vehicle treatment group, while it had no effect on blood glucose level. About renal functions, both of UACR and serum creatinine were significantly lower in D-532 treatment group. Importantly, renal tissues such as glomeruli and renal tubules were preserved in D-532 treatment group compared with in vehicle treatment group. In vivo as well, D-532 suppressed ChREBP target genes mRNA expression, and RNA-sequence analysis with mice renal cortex RNA at the end of D-532 administration experiment indicated that D-532 suppressed excessive oxidative stress through the ChREBP activation under diabetes.

Conclusion:

These results indicate that ChREBP suppression is effective on DN rodent model, and ChREBP activity inhibitor D-532 may have potential as a new treatment for DN in the future.
PS-BPB06-4: EXERCISE TRAINING AMELIORATES RENAL OXIDATIVE STRESS IN RATS WITH CHRONIC KIDNEY DISEASE
Authors: Yamakoshi Seiko , Nakamura Takahiro , Mori Nobuyoshi , Kohzuki Masahiro , Mori Takefumi , Ito Osamu
Published: January 2023

Background:

The renoprotective effects of long-term exercise in chronic renal failure have been reported. However, the details of the mechanism remains unclear. This study investigated the renal protective effect of long-term exercise on renal oxidative stress in a rat model of chronic renal failure.

Methods:

Male Sprague-Dawley rats were divided into Sham, 5/6 nephrectomy + sedentary, and 5/6 nephrectomy + exercise training groups. 5/6 nephrectomy + exercise training group underwent treadmill running for 12weeks. Systolic blood pressure and urinary protein were measured every 2 weeks. Renal function, renal histology and oxidative stress-related factors were examined after completion of the exercise protocol.

Results:

5/6 nephrectomized rats exhibited hypertension, proteinuria, renal dysfunction glomerulosclerosis, renal interstitial fibrosis, and exercise training ameliorated them. 5/6 nephrectomy increased urinary malondialdehyde, renal cortical NADPH oxidase and xanthine oxidase activities and exercise training inhibited the 5/6 nephrectomy-increased urinary malondialdehyde, renal cortical NADPH oxidase and xanthine oxidase activities. Plasma malondialdehyde was not affected by 5/6 nephrectomy or exercise.

Conclusions:

In rats with chronic kidney disease, long-term exercise prevented hypertension, renal dysfunction, glomerulosclerosis, and renal interstitial fibrosis. Mechanism for the antihypertensive and renal protective effects of long-term exercise in chronic kidney disease may involve the reduction of renal oxidative stress.
PS-BPB06-6: RENAL INJURY ASSOCIATED WITH ARGININE VASOPRESSIN INDUCTION IN A RAT MODEL OF PREECLAMPSIA
Authors: Ramdin Sapna , Baijnath Sooraj , Naicker Thajasvarie , Govender Nalini
Published: January 2023

Objective:

Worldwide, preeclampsia (PE) accounts for 2–8% of pregnancies and is a main cause of maternal and fetal morbidity and mortality. In vivo murine studies confirm the use of arginine vasopressin (AVP) in replicating the clinical symptoms of PE development, suggestive of a role for PE prediction. The renal effects that AVP may have in PE remains elusive, hence this study aimed to explore the physiological effects of AVP on the kidneys of pregnant rats in an effort to determine its role in PE development.

Design and method:

Twenty-four female Sprague Dawley rats (non-pregnant saline, pregnant saline, non-pregnant AVP and pregnant AVP) were surgically implanted with ALZET mini osmotic pumps infusing with either saline or AVP for 18 days. Blood pressure measurements and urine samples were taken on gestational days (GD) 8, 14 and 18. Placental and fetal weights were recorded following sacrifice on GD18. Urinary levels of kidney injury molecule-1 (KIM-1) were determined using the rat kidney toxicity immunoassay panel 1 (#RKTX1MAG-37K). Immunohistochemistry was used to determine the expression of KIM-1 in rat kidney tissue.

Results:

Systolic [143.00 (139.67–144.00) mmHg vs 125.00 (124.00–127.00) mmHg; p < 0.05] and diastolic blood pressures [103.50 (100.33–106.88) mmHg vs 81.00 (80.00–87.00) mmHg; p < 0.001] were significantly higher in the pregnant AVP vs pregnant saline group. Urinary protein levels in pregnant AVP rats [0.45 (0.42–0.48) g/L] were significantly elevated in comparison to pregnant saline rats [0.24 (0.21–0.32) g/L; p < 0.05]. Pregnant AVP rats also demonstrated a significant decrease in individual placental [0.39 (0.31–0.46) g vs 0.59 (0.55–0.63) g; p < 0.05] and pup weights [1.57 (1.45–1.67) g vs 1.79 (1.70–1.85) g; p < 0.05] in comparison to pregnant saline rats. Urinary KIM-1 levels are significantly elevated in the pregnant AVP (0.53 ± 0.05 ng/mL) vs pregnant saline group (0.17 ± 0.02 ng/mL; p < 0.001). Qualitative analysis revealed that KIM-1 expression is markedly upregulated on the apical membrane surface of epithelial cells of the proximal convoluted tubules in AVP treated rats.

Conclusions:

AVP infusion increases blood pressure, urine protein levels and urinary and tissue expression of KIM-1. Our findings indicate that there is significant kidney injury associated with AVP treatment, indicative of the utility of this model in studying the mechanisms driving renal damage in PE.
PS-BPB06-9: PREVALENCE OF KCNJ5 MUTANT ALDOSTERONE-PRODUCING ADENOMAS AMONG MALAYSIAN PATIENTS WITH PRIMARY ALDOSTERONISM
Authors: Rizam Nurfatin Syazreen Mohd , Mustangin Muaatamarulain , Tan Geok Chin , Sukor Norlela , Saparuddin Fatin , Pauzi Fatin Athirah , Nazri Siti Khadijah Syed Muhammad , Nasruddin Azraai Bahari , Azizan Elena Aisha
Published: January 2023

Objective:

To verify the prevalence of KCNJ5 mutant aldosterone-producing adenomas (APAs) among primary aldosteronism (PA) patients from a tertiary center in Malaysia

Design and method:

A total of 80 adrenal samples from adrenalectomies performed between 2010 up to 2020 at Hospital Putrajaya, Malaysia were included in this study. Of the adrenals blocks received, DNA samples from 74 APAs identified by CYP11B2 immunohistochemistry (IHC) staining were sent for KCNJ5 mutation analysis. Gender, ethnicity and age at adrenalectomy were compared between genotypes. Data were statistical analyzed using the Fisher Exact test.

Results:

28 of the 74 (37.8%) APAs had a KCNJ5 mutation. The most common KCNJ5 variant was G151R (50%), followed by L168R (39.3%) and T158A (10.7%). Of all the mutant APAs, 19/28 (67.9%) were from female PA patients whereas among controls 26/46 (56.5%) were females. No significant association between KCNJ5 mutation status and gender were found. The majority of mutant APAs were from Malay PA patients (n = 17), followed by Chinese PA patients (n = 11) which follows the whole cohort demography. Thus, ethnicity was not found to be significantly associated with KCNJ5 mutation status. In this cohort, no significant differences in age at of adrenalectomy by KCNJ5 mutation status was seen. Adrenalectomy procedure was performed most frequent between the age of 37–46 years old (n = 30), followed by 47–56 (n = 16), 26–35 (n = 12), 57–66 (n = 11) and 67–76 (n = 2); in 3 patients this information was not known.

Conclusions:

A similar prevalence of KCNJ5 mutant APAs as found in a previous study performed at the National University of Malaysia Medical Centre between 2000 and 2015 verifies the lower prevalence of KCNJ5 mutant APAs in Malaysian PA patients (37.8%) compared to that observed in other Oriental cohorts (up to 70%). Although, the majority of mutant APAs were from female PA patients (67.9%), no gender bias according to KCNJ5 mutation status was found as the majority of PA patients in this cohort were females (60.8%).
PS-BPB06-10: DISTRIBUTION OF CACNA1D SOMATIC MUTATIONS AMONG CYP11B2 IMMUNOHISTOCHEMISTRY-GUIDED SAMPLES OF ALDOSTERONE-PRODUCING ADENOMA (APA)
Authors: Pauzi Fatin Athirah , Azizan Elena Aisha , Mustangin Muaatamarulain , Tan Geok Chin , Sukor Norlela , Ryska Ales , Ceral Jiri , Solar Miroslav , Nasruddin Azraai Bahari
Published: January 2023

Objectives:

Past studies in Afro-Caribbean primary aldosteronism (PA) patients have found CACNA1D mutations to be the most common aldosterone-driving mutation among their cohort; a striking difference to Caucasian and Oriental cohorts where KCNJ5 mutations are reported to be the most common. However not many studies had used aldosterone synthase (CYP11B2) immunohistochemistry (IHC)-guided sequencing approach of the whole CACNA1D gene, performing instead unguided adenoma sampling and hotspot sequencing. We thus performed CYP11B2 IHC-guided whole gene sequencing on APAs from an Oriental cohort (Malaysia) and a Caucasian cohort (Czech Republic) for comparison.

Design and method:

Surgically removed adrenals from 100 Malaysian and Czech PA patients with unilateral disease were studied. Genomic DNA was isolated from adenoma FFPE sections that had a positive CYP11B2 IHC staining and somatic mutations analysis were performed by next generation sequencing (NGS) of known aldosterone-driver genes. Predicted pathogenic mutations were verified by Sanger Sequencing.

Results:

Out of 100 FFPE samples sent for NGS, 36 were excluded due to the quality of the data. In total 58 of 64 APAs (87.5%) had a pathogenic aldosterone-driver somatic mutation. KCNJ5 mutations were the most common (n = 37, 57.8%), followed by CACNA1D (n = 11, 20.3%), ATP1A1 (n = 6, 9.4%), ATP2B3 (n = 3, 4.7%) and CTNNB1 (n = 2, 3.1%) mutations. Of particular interest, one APA sample had two pathogenic somatic mutations, CACNA1D p.Val1373Met and CTNNB1: p.Thr41Ala. SIFT and Polyphen2 scores predicted both mutations to be deleterious. Both mutations have previously been reported separately to cause APAs. The CACNA1D/CTNNB1 mutant APA, was solitary, with homogeneous CYP11B2 expression while negative for CYP17A1 expression. The APA was adrenalectomized from a 53-year-old male Czech patient. The majority of patients with APAs harboring CACNA1D mutations were male (n = 8, 72.7%) and from the Czech cohort (n = 7, 64%). There were no significant differences in age at adrenalectomy between patients harboring a CACNA1D mutant APA (45 ± 3) and those with a non-CACNA1D mutant APA (48 ± 2).

Conclusions:

Solitary ZG APAs can harbor both a CACNA1D and a CTNNB1 mutation. The prevalence of CACNA1D in this CYP11B2-IHC guided study is consistent with previous results reported in Oriental and Caucasian cohorts. Although CACNA1D is not the top genetic alteration responsible for APAs in these cohorts, CACNA1D may still be a potential key player in idiopathic hyperaldosteronism and aldosterone-producing micronodules. Thus further study is required to characterize the distribution of CACNA1D pathogenic mutations in bilateral PA.
PS-BPB07-2: AFFERENT RENAL NERVES CONTRIBUTE TO LEFT VENTRICULAR DYSFUNCTION THROUGH HYPOTHALAMIC VASOPRESSIN-MEDIATED CENTRAL SYMPATHOEXCITATION IN HYPERTENSIVE HEART FAILURE MODEL
Authors: Ikeda Shota , Shinohara Keisuke , Kashihara Soichiro , Matsumoto Sho , Yoshida Daisuke , Nakashima Ryosuke , Ono Yoshiyasu , Tsutsui Hiroyuki
Published: January 2023

Objective:

Afferent renal nerves may contribute to the development and progression of heart failure, which, however, has not been well investigated. This study aimed to examine whether the afferent renal nerve input contributes to hypertensive cardiac dysfunction.

Design and method:

Male Dahl salt-sensitive rats were fed 0.3% low-salt diet (LS) or 8% high-salt diet (HS) from 6 weeks of age. HS-fed rats were divided into selective afferent renal denervation (HS-ARDN) or sham operation (HS-Sham) at 9 weeks. The effects of ARDN on central sympathetic regulation were investigated at 12 weeks. In another cohort, its effects on the cardiac phenotypes were evaluated at 16 weeks, with adding total renal denervation (HS-TRDN) group.

Results:

At 12 weeks, systolic blood pressure (SBP) measured by tail-cuff method was significantly increased in HS-Sham compared to LS, while there was no significant difference between HS-Sham and HS-ARDN. Plasma norepinephrine concentration was increased in HS-Sham compared to LS and was attenuated in HS-ARDN. The c-Fos expression, a neuronal activation marker, in presympathetic neurons within hypothalamic paraventricular nucleus (PVN) and rostral ventral lateral medulla (RVLM) was also increased in HS-Sham and attenuated in HS-ARDN, which was in parallel with the c-Fos expression in vasopressin-expressing neurons within PVN. At 16 weeks, SBP was similarly increased in HS-Sham, HS-ARDN, and HS-TRDN compared to LS. Although the parameters of left ventricular (LV) hypertrophy, such as LV wall thickness by echocardiography and LV weight, were significantly increased in HS-Sham, HS-ARDN, and HS-TRDN compared to LS, there was no significant difference between HS-fed three groups. Echocardiographic LV fractional shortening was significantly decreased in HS-Sham compared to LS, and was attenuated to the same extent in HS-ARDN and HS-TRDN. Histological LV fibrosis determined by Masson trichrome staining and mRNA expression of ANP and CTGF in LV were significantly increased in HS-Sham compared to LS; these changes were similarly ameliorated in both HS-ARDN and HS-TRDN.

Conclusion:

The input from renal afferent nerves contributes to sympathoexcitation and cardiac dysfunction independently of SBP in Dahl salt-sensitive hypertensive heart failure model. Vasopressin production in PVN may be involved in the renal afferents-mediated sympathetic regulation.
PS-BPB07-4: LONG-TERM PATHOLOGICAL CHANGES IN ABLATED RENAL NERVES FOLLOWING CATHETER-BASED RADIOFREQUENCY RENAL DENERVATION IN A NORMOTENSIVE SWINE MODEL
Authors: Schlaich Markus , Tunev Stefan , Sharp Andrew , Lee David P , Finn Aloke , Trudel Julie , Kandzari David E
Published: January 2023

Objective:

Randomized, sham-controlled clinical trials with catheter-based radiofrequency (RF) renal denervation (RDN) have demonstrated safety and efficacy of the procedure through 3 years. However, pre-clinical reports of functional renal nerve regrowth are inconsistent. In this analysis in normotensive swine, we assessed the specificity of RF ablation around the renal arteries out to 28 days and quantified the long-term pathophysiological outcomes through 180 days.

Design and method:

RF ablations were delivered to the renal arteries of normotensive swine using the Symplicity SpyralTM catheter (Medtronic, Santa Rosa, CA, USA). For the tissue specificity analysis, RF RDN was performed in 14 swine, and serial sections of the renal artery were harvested from 6 pigs at day 7, and 8 pigs at day 28. Surrounding anatomic tissues of the treated areas (including ureter, kidney, lymph nodes, skeletal muscle, adrenal glands, and renal nerves) were evaluated in response to RDN using a semi-quantitative scoring system (Figure). To assess durability, RF RDN was performed in a separate group of 14 normotensive swine, with an additional 14 pigs serving as control. Serial sections of renal artery were harvested at day 7 (n = 6 animals per group) and day 180 (n = 8 per group) post-procedure. In addition, matrixed samples from renal cortex were collected for each pig in both studies and subjected to bioanalytical quantification of cortical norepinephrine (NE) levels and immunohistochemical analysis of renal cortical axon density.

Results:

At 7 and 28 days, cell disruption in non-target perivascular tissues (ureter, kidney, lymph nodes, skeletal muscle and adrenal glands) were rare/absent, whereas renal artery nerve disruption and axonal loss were sustained through 28 days (Figure). Through 180 days, nerve atrophy due to axonal destruction was apparent within and downstream to the ablation regions. Consistent nerve necrosis at the ablation sites was evident at day 7, but not day 180. Fibrosis was confined to the ablated regions at 7 and 180 days. Renal cortical axon density and corresponding cortical NE levels were significantly reduced at 7 and 180 days in the RF RDN group vs. control.

Conclusion:

RF RDN specifically disrupted the renal nerves around the renal artery leading to downstream axonal loss in the renal cortex. Renal cortical axon density, associated NE reduction, and nerve atrophy due to axonal destruction were sustained through 180 days post-RDN in a normotensive swine model. These results support the existing clinical evidence of specific and durable effects of RF RDN in lowering blood pressure.
PS-BPB07-5: HYPERTENSIVE STIMULI PROMOTE NEUROINFLAMMATION AND COGNITIVE IMPAIRMENT IN A PRESSURE-DEPENDENT MANNER
Authors: Dinh Quynh Nhu , Selvaraji Sharmelee , Lo Cecilia , Vinh Antony , Kim Hyun Ah , Chrissobolis Sophocles , Drummond Grant R , Arumugam Thiruma , Sobey Christopher G , De Silva T Michael
Published: January 2023

Objective:

Hypertension greatly increases the risk for cognitive impairment, and is associated with inflammation of the brain. However, it is unclear to what extent there is neuroinflammation during hypertension and whether this promotes cognitive impairment. This study aimed to quantify neuroinflammatory and cognitive changes caused by administration of angiotensin II, and determine whether this is blood pressure-dependent.

Design and method:

Male C57Bl/6 mice were administered vehicle (saline; n = 33) or angiotensin II (0.7 mg/kg/d, n = 34) for 14 d via an osmotic minipump. A subset of mice also received hydralazine hydrochloride (50 mg/kg; n = 38) in their drinking water for 14 d after minipump implantation. Systolic blood pressure (SBP) was measured using tail-cuff; inflammation was assessed using flow cytometry and RNA sequencing; and recognition memory was evaluated using the novel object recognition test.

Results:

Angiotensin II infusion increased SBP and promoted accumulation of neutrophils, monocytes, T cells and B cells in the brain compared to vehicle infusion which was blunted by co-administration of hydralazine (n = 7–8, P < 0.05). Angiotensin II infusion also impaired recognition memory and this effect could be prevented by co-treatment with hydralazine (n = 11–12, P < 0.05). Angiotensin II infusion promoted differential expression of 1194 genes in whole brain hemisphere and 634 genes in hippocampus compared to vehicle infusion. Out of these genes, 295 in whole brain hemisphere and 125 in hippocampus were inflammation/immune-related (n = 6, P-adjusted < 0.05). Angiotensin II caused downregulation of genes involved in cognitive function and memory consolidation (Dusp1, Arc) and upregulation of genes involved in immune cell function, regulation and migration (H2-Ab1, H2-Q6, Il17rc, Fcrlb, Ltb, Ccl8, Itga9) in whole brain hemisphere (n = 6, P-adjusted < 0.05). In the hippocampus, a key region in learning and memory, angiotensin II promoted upregulation of genes involved in immune cell proliferation and differentiation (Il12rb2, Acvr1b, Sema3c) and apoptosis (Plxna3, Anxa11, Hyal1, Axin1) (n = 6, P-adjusted < 0.05). Co-treatment with hydralazine altered expression of all of these genes suggesting possible reversal of the transcriptomic changes induced by angiotensin II infusion (n = 6, P-adjusted < 0.05).

Conclusion:

Angiotensin II promotes neuroinflammation and cognitive impairment in a blood pressure-dependent manner. Co-treatment with hydralazine may reverse some of the transcriptomic changes induced by angiotensin II. Chronic neuroinflammation may be a contributing factor to cognitive impairment during hypertension and could be modulated by blood pressure reduction.
PS-BPB07-6: NOVEL HUMORAL AND NEURONAL PATHWAYS FOR DIRECT BLOOD PRESSURE CONTROL BY MACULA DENSA CELLS
Authors: Gyarmati Georgina , Shroff Urvi Nikhil , Deepak Sachin , Izuhara Audrey , PetiPeterdi Janos
Published: January 2023

Objective:

According to traditional physiological knowledge, macula densa (MD) cells of the juxtaglomerular apparatus (JGA) in the kidney maintain body fluid homeostasis and blood pressure (BP) indirectly by controlling renal and glomerular hemodynamics and renin secretion based on local sensory inputs. However, recently identified microanatomical (long, axon-like basal cell processes, secretory vesicles) and functional (high level of protein synthesis) features of MD cells suggest they perform non-traditional functions. The present study addressed the hypothesis that MD cells can directly and acutely control distant cardiovascular organ functions and blood pressure via novel secreted hormones and communications with renal nerves.

Design and method:

A comprehensive research approach used transgenic mouse models (MD-GFP, MD-Ai27 mice that selectively express the depolarizing channel rhodopsin ChR2 in MD cells), intravital multiphoton imaging, systemic BP monitoring via the cannulated carotid artery, whole mount kidney imaging and 3D analysis, and single-cell transcriptomic analysis.

Results:

Bulk and single cell MD transcriptome analysis identified the high expression of secreted angiogenic and vasoactive factors (Pappa2, Vash2, Cyr61, Aard, Pamr1) and synaptic proteins (Unc5d, Tenm2, Hpcal4, Nsg2, Syp, Syt5, Grin2c, Gabrr2, Begain) with their altered kidney and plasma levels in disease and MD gain and loss-of-function conditions. Immunohistochemistry and 3D tissue volume rendering of optically cleared whole-mount MD-GFP kidneys identified the high MD expression of synaptophysin and the close anatomical association between MD cell basal processes and tyrosin-hydroxylase (TH)+ or calcitonin gene-related peptide (CGRP)+ sympathetic and sensory nerve endings. Optogenetic MD stimulation was induced by blue light (470 nm) exposure of the whole left kidney of MD-Ai27 mice, which maneuver acutely and reversibly increased systemic BP by 22 ± 5mmHg (P < 0.05, n = 5).

Conclusions:

MD cells secrete hormones that target the vasculature and form synapses with sympathetic and sensory renal nerves. These novel MD and JGA pathways may be involved in direct BP control and represent exciting future therapeutic opportunities for renal and cardiovascular disease and hypertension.
PS-BPB08-3: OFF-TARGET EFFECTS OF SLEEVE GASTRECTOMY ON BLOOD PRESSURE INDEPENDENT OF BODY WEIGHT REDUCTION IN DIET-INDUCED OBESITY RATS: ROLE OF CHOLECYSTOKININ
Authors: Yoshida Yuichi , Gotoh Koro , Masaki Takayuki , Ozeki Yoshinori , Miyamoto Shotaro , Okamoto Mitsuhiro , Shibata Hirotaka
Published: January 2023

Objective:

Sleeve gastrectomy (SG) is currently the most selected surgical bariatric procedure for weight loss by shrinking the stomach. SG has been reported to improve glucose metabolism by altering gastrointestinal hormones, but the details of why blood pressure improves are not known. The purpose of this study is to elucidate the mechanism of why SG lowers blood pressure in diet induced obese rats.

Design and method:

Male Sprague Dawley (SD) rats were subjected to surgical [sham operation (SO) or SG] and dietary interventions [fed a normal diet or high fat diet (HFD) ad libitum or pair fed (PF)]. Diet induced obese rats fed 60% HFD for 12 weeks. Systolic blood pressure (SBP), urinary sodium excretion, and gastrointestinal hormones and renin angiotensin system in renal tissue were examined 4 weeks after surgery. In addition, cholecystokinin (CCK) was administered intraperitoneally to SD rats and evaluated for changes in blood pressure and renal tissue renin angiotensin system.

Results:

Both SG and PF treatments reduced body weight compared with SO. SG exhibited a reduction in SBP compared with PF, which was associated with a reduction in renal renin (536 ± 48.5 U/g kidney and 1385 ± 55.1 U/g kidney, p < 0.01), angiotensin II (Ang II) (48.2 ± 5.8 ng/g kidney and 258.7 ± 16.9 ng/g kidney, p < 0.01), and catechol O methyltransferase (COMT) levels (0.68 ± 0.05 /αtub and 1.34 ± 0.13/αtub, p < 0.01). SG increased plasma CCK levels compared with PF (p < 0.01 for each), whereas GLP1 and PYY were not changed in fasting. SD rats after CCK administration showed decreased SBP (107.7 ± 2.3 mmHg and 118.0 ± 1.8 mmHg, p < 0.01), COMT (0.49 ± 0.04 /GAPDH and 1.11 ± 0.19/GAPDH, p = 0.02), renin (967 ± 71.3 U/g kidney and 1394 ± 90.8 U/g kidney, p < 0.01), and Ang II (598 ± 76.7 ng/g kidney and 874 ± 86.6 ng/g kidney, p = 0.02) in the kidney compared to controls.

Conclusions:

The SG reduced SBP, at least in part, through suppression of the sympathetic nerve action by elevation of CCK which is followed by suppression of the intrarenal renin angiotensin system.

Conclusions:

CCK may have a blood pressure lowering effect, and further research is needed to determine whether CCK can be used as a new antihypertensive agent in the future.
PS-BPB08-7: MECHANISMS OF MINERALOCORTICOID RECEPTOR-ASSOCIATED HYPERTENSION IN DIABETES MELLITUS: ROLE OF O-GLCNAC MODIFICATION
Authors: Jo Rie , Shibata Hirotaka , Kurihara Isao , Yokota Kenichi , Kobayashi Sakiko , Murai-Takeda Ayano , Mitsuishi Yuko , Hayashi Takeshi , Nakamura Toshifumi , Itoh Hiroshi
Published: January 2023

Objective:

Recently administration of non-steroidal mineralocorticoid receptor (MR) antagonist with maximal tolerable doses of the renin-angiotensin system inhibitors significantly reduced renal as well as cardiovascular composite outcomes in patients with type 2 diabetes and chronic kidney diseases. However, molecular mechanisms of how MR is activated are not understood. Post-translational modification of O-linked-N-acetylglucosamine (O-GlcNAc) has emerged as an essential glucose-sensing mechanism. The aim of this study was to clarify whether MR is modified by O-GlcNAc and figure out the significance of this modification on MR in the context of diabetes mellitus.

Design and method:

1) The MR transcriptional activities were investigated in reporter assays and real time RT-PCR. Levels of expression of MR were examined with western blot analysis. Cells were cultured in 5 mM or 30 mM glucose conditions and additionally treated with PUGNAc, an O-GlcNAcase inhibitor, or 6-diazo-5-oxonorleucine (DON), a glucosamine-fructose-6-phosphate amidotransferase inhibitor. 2) O-GlcNAc modification of MR was investigated with coimmunoprecipitation assays. We mapped actual O-GlcNAc sites on MR by liquid chromatography-tandem mass spectrometry and generated several MR mutants. 3) We evaluated the expression levels of MR and the total O-GlcNAc levels in kidneys of 9-week-old db/db mice. We also intraperitoneally administered 3 g/kg DON to them for five days and evaluated the same subjects.

Results:

1) Treatments with either 30 mM glucose or PUGNAc enhanced 10–10 M aldosterone-mediated MR transactivation by 2-fold and increased endogenous SGK1 mRNA levels. These changes were accompanied by increased expression levels of MR protein. In contrast, treatment with DON lowered a reporter activity and the expression levels of MR. 2) Coimmunoprecipitation assays showed that MR was modified by O-GlcNAc and this modification of MR was increased by 30 mM glucose and PUGNAc treatment. The MR mutant of multiple O-GlcNAc sites (S295A/S296A/S298A/S299A) showed the decrease of its O-GlcNAc level, which was accompanied by lowering their expression levels and transcriptional activities by 40%. 3) The levels of MR protein increased by 2.5-fold in parallel with O-GlcNAc levels in kidneys of db/db mice. In addition, mRNA levels of SGK1 increased by 2-fold in db/db mice. Conversely, administration of DON decreased the expression levels of MR parallel to the total O-GlcNAc levels in kidneys.

Conclusions:

We showed for the first time that MR is a target for O-GlcNAc modification. High glucose conditions enhance O-GlcNAc modification of MR, which are associated with increased MR levels and its transcriptional activities in vitro and in vivo. The epigenetic modification of MR may account for the pathological MR activation in diabetes mellitus.
PS-BPB09-4: ARTERIAL SITES AND SEX DIFFERENCES IN ENHANCING VASORELAXATION RESPONSE BY PERIVASCU-LAR ADIPOSE TISSUE IN METABOLIC SYNDROME RATS
Authors: Kagota Satomi , Futokoro Risa , Fumoto Kana Maruyama , Shinozuka Kazumasa
Published: January 2023

Objective:

Regulation of arterial tone by perivascular adipose tissue (PVAT) differs with sex. We have demonstrated that PVAT compensates vascular tone when vascular dysfunc-tion occurs in mesenteric arteries of male SHRSP. Z-Leprfa/IzmDmcr (SPZF) rats, but such compensation function disappears in the late stage of metabolic syndrome (MetS). In con-trast, the favorable effects of PVAT remain in female SPZF even after the effects disappear in age-matched males.

Design and method:

Therefore, in this study, we investigated whether the sex differences in PVAT response are observed in another arterial sites in SPZF and in another MetS model, SHR/NDmcr-cp (CP) rats as well. Renal arteries were isolated from male and female SPZF rats and CP rats at 23 weeks of age. Ring preparations with and without PVAT were made. Vasodilation and mRNA transcript levels in PVAT were examined using organ bath methods and quantitative real-time PCR, respectively.

Results, and Conclusions:

In renal arteries without PVAT of SPZF and CP rats, acetylcholine-induced relaxations were smaller in female than in male. The presence of PVAT increased the relaxations in CP rats only. In contrast, sodium nitroprusside-induced relaxations, nevertheless presence or ab-sence of PVAT, were smaller in female than in male in SPZF rats, but sex differences were not observed in CP rats. Apelin mRNA levels in female CP rats were the highest among groups. The enhancements of acetylcholine-induced relaxations by PVAT were positively correlated with apelin mRNA levels in PVAT. These results demonstrated that sex difference in enhancing vasorelaxation response by PVAT in renal arteries differs to that in mesenteric arteries in SPZF rats at the same age and to that in another MetS model strain, CP rats, at the same age and arterial sites. This discrepancy observed in female rats between two MetS strains may be associated with the difference in arterial dysfunction mechanism; namely, impairment of only endothelial nitric oxide production in CP rats, and deterioration of re-sponse to nitric oxide in smooth muscle in SPZF rats. Furthermore, apelin levels of PVAT may be involved in the appearance of modulation of PVAT on arterial tone.
PS-BPB10-1: INVOLVEMENT OF RENAL TUBULAR NFAT5 IN THE REGULATION OF BLOOD PRESSURE AND BODY FLUID VOLUME
Authors: Izumi Yuichiro , Hiramatsu Akiko , Eguchi Koji , Matsuo Naomi , Deng Qinyuan , Inoue Hideki , Nakayama Yushi , Nonoguchi Hiroshi , Aramburu Jose , Rodriguez Cristina Lopez , Kakizoe Yutaka , Adachi Masataka , Kuwabara Takashige , Mitsuyama Shokei Kim , Mukoyama Masashi
Published: January 2023

Objective:

Nuclear factor of activated T-cells 5 (NFAT5) is an osmoresponsive transcription factor that activates osmoprotective genes to protect cells against hypertonicity. Renal tubular cell-specific NFAT5 knockout (KO) mice have been shown to exhibit polyuria, hypernatremia, low urinary sodium excretion, and increased expression of the epithelial sodium channel (ENaC) in the kidney. In the present study, regulation of blood pressure (BP) as well as that of body fluid volume were examined in the wild type (WT) mice and NFAT5 KO mice.

Methods:

The BP of KO mice and WT mice was measured using radiotelemetry method. The mice were fed with regular salt diet (RSD, 0.3% NaCl chow) or high salt diet (HSD, 8% NaCl chow) followed by the treatment with amiloride, an ENaC blocker. To assess body fluid volume, the plasma renin activity (PRA) and the plasma aldosterone concentration (PAC) were measured at the condition of RSD and HSD, together with plasma arginine vasopressin (AVP) and hematocrit (Ht) levels.

Results:

Compared to WT mice, the mean BP was significantly higher in KO mice both at the condition with RSD and that with HSD in dark period, but not in light period. Treatment with amiloride significantly decreased the mean BP in KO mice with HSD to the level of WT mice with HSD in dark period. PRA, PAC, and AVP levels were not significantly different between in WT and KO mice with RSD. HSD significantly decreased PRA and PAC and tended to increase the AVP level both in WT and KO mice. The Ht level was significantly lower in KO mice than in WT mice regardless of RSD or HSD (WT with RSD, 42.6 ± 0.5%; WT with HSD, 42.8 ± 0.6%; KO with RSD, 38.3 ± 0.4%; KO with HSD, 39.0 ± 0.4%, n = 4).

Conclusions:

The results suggest that the renal tubular cell-specific NFAT5 KO mice exhibit excess body fluid retention induced by sodium reabsorption through ENaC, leading to salt-sensitive hypertension. Such phenotypes seem to be independent of the systemic renin-angiotensin-aldosterone and AVP systems. Renal tubular NFAT5 could integrate urine concentration and sodium reabsorption in the kidney, thus preventing salt-dependent hypertension.
PS-BPC01-2: PREDICTORS AND EFFECTS OF COMORBID ATRIAL FIBRILLATION IN HYPERTENSIVE PATIENTS DUE TO PRIMARY ALDOSTERONISM
Authors: Sakaguchi Shintaro , Okamoto Ryuji , Inoue Chisa , Akao Masaharu , Kamemura Kohei , Takeda Yoshiyu , Ohno Youichi , Inagaki Nobuya , Rakugi Hiromi , Tanabe Akiyo , Tamura Kouichi , Fujita Satoshi , Yano Yutaka , Dohi Kaoru , Naruse Mitsuhide
Published: January 2023

Objective:

The incidence of atrial fibrillation (AF) and risk of cardiovascular events are reportedly higher in patients with primary aldosteronism (PA) than those with essential hypertension. This study sought to determine independent predictors of comorbid AF and cardiovascular events in PA patients after PA treatment.

Design and method:

This nationwide registration study included PA patients more than 20 years old. Incident cardiovascular events were observed with a mean follow-up of approximately 3 years. A total of 3,654 patients with PA were included at the time of analysis.

Results:

Prevalence of AF was 2.4%. PA patients with AF were older, more frequently male and had longer duration of hypertension than those without AF. No significant difference in basal plasma and adrenal venous aldosterone concentration, renin activity, potassium concentration, confirmatory tests of PA, laterality or surgery rate were seen between groups. Logistic regression analysis showed age, male sex, cardiothoracic ratio, past history of coronary artery disease and heart failure were independent determinants for AF. PA patients with AF showed a higher frequency of cardiovascular events than those without AF (P < 0.001). Multivariate Cox analyses demonstrated AF in addition to duration of hypertension, older age, male sex, hypokalemia and surgery as independent prognostic factors for cardiovascular events (hazard ratio [HR] 1.866, 95% confidence interval [CI] 1.024–3.401, P < 0.05; HR 1.022, 95%CI 1.006–1.039, P < 0.01; HR 1.424, 95%CI 1.043–1.943, P < 0.05; HR 1.027, 95%CI 1.012–1.042, P = 3.2 × 10–4; HR 1.726, 95%CI 1.226–2.430, P < 0.005; HR 1.458, 95%CI 1.041–2.041, P < 0.05, respectively). After PA treatment, incidence of cardiovascular events did not differ significantly between PA patients with AF and essential hypertension patients with AF from the Fushimi registry during follow up after adjusting age, sex and prevalence of hypertension.

Conclusion:

Early diagnosis of PA may prevent AF and other cardiovascular events in PA patients by shortening the duration of hypertension.
PS-BPC01-5: PROGNOSTIC IMPLICATIONS OF MACHINE LEARNING-DERIVED ECHOCARDIOGRAPHIC PHENOTYPES IN ASYMPTOMATIC COMMUNITY HYPERTENSIVE POPULATIONS
Authors: Cai Anping , Zhou Dan , Zhou Yingling , Tang Songtao , Feng Yingqing , Nie Zhiqiang
Published: January 2023

Objective:

Echocardiogram is commonly used to evaluate cardiac remodeling in hypertension (HTN). However, study on echocardiographic phenotypes and their prognostic implications in HTN is limited. This study aimed to elucidate these aspects.

Method:

A total of 1881 community hypertensive patients without overt cardiovascular disease and severe renal disease (mean age 62.8 years, women 57.9%) were included. Using Two-Step cluster analysis with four conventional echocardiographic variables, two clusters with distinct echocardiographic phenotypes were identified.

Result:

The Cluster 1 (namely mild-remodeling HTN; n = 1492) had low prevalence of enlarged left atrium (LA; 0.9%) and left ventricular hypertrophy (LVH; 16.2%) and better LV diastolic function. They were younger and more likely to be men and had lower comorbid burden. The Cluster 2 (namely severe-remodeling HTN; n = 389) had higher prevalence of enlarged LA (26.0%) and LVH (83.0%) and worse LV diastolic function. They were older and more likely to be women and had higher comorbid burden. After a median follow-up of 4.2 years, compared to the Cluster 1, the Cluster 2 had higher incidence of cardiovascular (4.1% vs 1.7%; P = 0.006) and all-cause (9.8% vs 4.8%; P < 0.001) death, with adjusted hazard ratio (HR) of 2.80 (95% confidence interval [CI] 1.39–5.62; P = 0.004) and 2.04 (95% CI 1.32–3.14; P < 0.001) respectively. Kaplan-Meier curve plotted the cumulative incidence of cardiovascular and all-cause mortality in the Figure 1 Panel A and B.

Conclusion:

In conclusion, these findings indicate that the conventional echocardiographic variables-based algorithm could be helpful to identify asymptomatic community hypertensive patients at risk for cardiovascular and all-cause death. Further studies are needed to develop and validate phenotype-specific prevention and intervention strategies in HTN.
PS-BPC02-1: PCSK-9-INHIBITOR THERAPY IMPROVES ENDOTHELIAL FUNCTION IN HIGH-RISK PATIENTS WITH CARDIOVASCULAR DISEASE
Authors: Kannenkeril Dennis , Bosch Agnes , Kolwelter Julie , Striepe Kristina , Pietschner Robert , Ott Christian , Schiffer Mario , Achenbach Stephan , Schmieder Roland E
Published: January 2023

Background:

Impaired endothelial function predicts cardiovascular events. The aim of this study was to analyse the effect of evolocumab on endothelial function in patients with cardiovascular disease and on obligatory statin therapy.

Methods:

This was a prospective, double-blinded, randomized, controlled, single center study (NCT03626831) including patients with cardiovascular disease (diagnosis of coronary artery disease, non-hemorrhagic stroke, transient ischemic attack or symptomatic peripheral artery disease) and treated with statins, similar to the inclusion criteria of the FOURIER study. Patients were consecutively randomized (1:1) to either evolocumab treatment (420 mg was administrated in monthly intervals, twice during the study) or placebo. All patients underwent examination of endothelial function at baseline, and after 1, 4 and 8 weeks of treatment by a semi-automatic high-resolution ultrasound system (UNEX EF 18G; Unex Co., Nagoya, Japan). Endothelial function parameter such as flow-mediated vasodilation (FMD; vasodilator responsiveness), low flow-mediated vasocontriction (L-FMC; vasoconstrictor responsiveness) and vasoactive range (VAR; total vasomotor responsiveness) were measured.

Results:

103 patients with a mean age of 66.2 (±7.7) years and a mean LDL-cholesterol of 98.2 (±19.1) mg/dl completed the study. 83.5% of the study population was known to have coronary artery disease. Endothelial function parameter (VAR) increased after 8 weeks of treatment with evolocumab compared to baseline (p = 0.034), whereas there was no significant change at 1 and 4 weeks after treatment. Moreover, an improvement in VAR from baseline at week 8 was found with evolocumab compared to placebo (p = 0.045).

In a subgroup analysis, in patients with age < 67 years, lower systolic blood pressure (< 125 mmHg) or higher baseline LDL-cholesterol (> 95 mg/dl), significant evolocumab treatment effect was found in VAR improvement (p = 0.006, p = 0.049 and p = 0.042, respectively) from baseline at week 8. No serious adverse event related to study medication occurred during the study.

Conclusion:

Our data indicate that endothelial function could be improved with evolocumab treatment in high-risk patients with preexisting cardiovascular disease and on statin therapy. Our results contribute to the mechanistic explanation why lower incidence of the cardiovascular composite endpoint has been demonstrated in the FOURIER study.
PS-BPC02-5: IMPACT OF LONGITUDINAL CHANGES IN METABOLIC SYNDROME STATUS OVER TWO YEARS ON 10-YEAR INCIDENT TYPE 2 DIABETES MELLITUS
Authors: Koh Kwang Kon
Published: January 2023

Background:

Temporal changes in MetS status over two years may modify the 10-year risk of incident diabetes mellitus (DM).

Methods:

A prospective cohort study was conducted in 7,317 adults aged 40–70 years without DM at baseline. Subjects were categorized into four groups based on repeated longitudinal assessment of metabolic syndrome (MetS) status over two years, as defined by ATP III guidelines: non-MetS, resolved MetS, incident MetS, and persistent MetS. The hazard ratio (HR) of new-onset DM during 10 years was calculated in each group using Cox models.

Results:

During the 10-year follow-up, 1,099 (15.0%) developed DM. Compared to the non-MetS group, the fully adjusted hazard ratios (95% confidence interval) for new-onset DM were 1.27 (1.01–1.61) in the resolved MetS group, 1.78 (1.43–2.22) in the incident MetS group, and 1.85 (1.52–2.26) in the persistent MetS group (P for trend < 0.001). The 10-year risk of DM in subjects with resolved MetS was attenuated compared to those with persistent MetS over two years (P < 0.001). The adjusted hazard ratio for 10-year incident DM gradually increased according to the number of MetS components two years later.

Conclusions:

Temporal changes in MetS status over two years influenced the 10-year risk of incident DM, suggesting that determining the status of MetS regularly and strictly controlling MetS are important in preventing DM.
PS-BPC02-7: RADIAL-DIGITAL PULSE WAVE VELOCITY: STIFFNESS OF SMALL CONDUIT ARTERIES INCREASES AFTER NITROGLYCERIN ADMINISTRATION
Authors: Fortier Catherine , Garneau Charles Antoine , Pare Mathilde , Obeid Hasan , Cote Nadege , Duval Karine , Goupil Remi , Agharazii Mohsen
Published: January 2023

Objective:

The alteration of the physiological stiffness gradient within larger vessels would increase the transmission of greater pulsatility to the microcirculation, thus explaining the damage to the pressure-sensitive organs. We aimed to describe the response of small conduit arteries of the hand (radial-digital PWV, rd-PWV) following the pharmacological alteration of the stiffness gradient using nitroglycerin.

Design and method:

Simultaneous application of piezoelectric sensors (Complior) at the level of the carotid (c), the radial artery (r) and the tip of the index finger (d) was used to calculate the rd-PWV by the following formula (rd-PWV = cd-cr direct distances/cd-cr transit times). Rd-PWV was obtained before and 4 minutes after a sublingual administration of 0.4 mg of NTG, in healthy adults (controls, n = 36) and patients with moderate chronic kidney disease (CKD, n = 30). Changes in arterial stiffness pre-post NTG and comparisons between the two groups were analyzed respectively with paired or independent samples t-tests.

Results:

Compared to controls, the CKD group was older (69 ± 15 years vs. 41 ± 20 years) and more hypertensive. Despite similar cd-PWV and cr-PWV, rd-PWV was significantly lower in CKD than in controls (3.6 ± 1.4 m/s vs. 4.5 ± 1.8 m/s, p = 0.024), but this difference faded (p = 0.145) after adjustment for age. Post NTG, brachial blood pressure decreased and heart rate increased similarly between groups (see Table 1). Cr-PWV decreased after NTG (both groups p < 0.001) while rd-PWV increased in controls (from 4.62 ± 1.49 m/s to 5.94 ± 2.29 m/s, p < 0.001) and in CKD (from 3.71 ± 1.60 m/s to 5.18 ± 2.12 m/s, p < 0.001), in a similar extent (p = 0.111 for interaction term). Cd-PWV significantly increased post-NTG only in the CKD group (p = 0.009).

Conclusions:

Again, we observed opposing changes in the regional stiffnesses of two distinct vascular territories following the administration of a vasodilator. Here, the two segments were mid-size and small conduit muscular arteries. This technique, adapted by our team for short peripheral arterial segments, may broaden our understanding of the consequences of the inversion of the stiffness gradient.
PS-BPC03-3: CARDIOVASCULAR EVENTS PREDICTION RELATED TO GENDER IN PATIENTS WITH HYPERTENSIVE EMERGENCIES IN 12-MONTHS FOLLOW-UP GREEK STUDY
Authors: Fragoulis Christos , Polyzos Dimitrios , Dimitriadis Kyriakos , Konstantinidis Dimitrios , Mavroudis Andreas , Leontsinis Ioannis , Kariori Maria , Drogkaris Sotirios , Tatakis Fotis , Manta Eleni , Siafi Eirini , Papakonstantinou Pantelis , Zammanis Ioannis , Thomopoulos Costas , Tsioufis Konstantinos
Published: January 2023

Objective:

Currently, epidemiological data on prognosis of patients with hypertensive crises (hC) are limited. The purpose of our study was to register the prevalence and clinical characteristics of patients with hypertensive urgencies (hU) and emergencies (hE) admitting to the emergency department (ED) and during hospitalization and a 12-month follow-up to record new end points in a Greek tertiary hospital.

Design and method:

The study population included patients reporting to the ED with acute rise in blood pressure (BP) (systolic BP >or = 180mmHg and/or diastolic BP >or = 120mmHg), categorizing as hE and hU depending on the presence or absence of acute hypertension-mediated target organ damage, respectively. In all patients the demographic and clinical information were registered for 12 consecutive months. In addition, there was a 12-month follow-up for new events.

Results:

Out of 38,589 patients assessed in the ED during a period of 12 months, 353(0.91%) had HC, out of whom 256(72.5%) had hU and 97(27.5%) had hE. The mean age of the hE patients was 73 ± 12years, 51.5% were males and 86.4% had a history of hypertension. During the study period 3 hE patients did not complete the follow-up. After stratification of hE population by gender, we created two subgroups, males and females, and analyzed the composite end point involving deaths or hospitalizations by cardiovascular causality. Men reported more composite events than women (21 vs 11 events, p = 0.042, hR for males 2.2, 95% CI 1.03–4.6). Cox regression models were adjusted for age, gender, cardiovascular disease, chronic kidney disease and diabetes mellitus.

Conclusions:

The results indicate male gender as an independent risk factor for deaths or hospitalizations caused by cardiovascular disease in patients with hE. Our registry highlights the commitment for a more intensive follow-up of hE patients, especially for men, and underlines the need for further research in this pathological entity.