Young Spontaneously Hypertensive Rats Research Articles

#4248 COMBINED EFFECT OF DEHYDRATION, HYPERTENSION, AND AGE ON RENAL FRAILTY

Abstract Background and Aims Renal frailty (RF) is a premorbid and, at least partly, modifiable condition arising from diminished renal functional reserve and defective adaptive response capacity predisposing to acute kidney injury (AKI). RF ensues from subclinical wear or distortion of the renal haemodynamic and tubular homeostatic responses that defend the renal excretory function from supervening circumstances (i.e., stressors such as drugs). In this work, we aimed to study the impact of RF generated by the combined effect of the hydration state, hypertension and ageing on the nephrotoxicity developed by a low dose of cisplatin in the rat. Method Young Wistar rats, young spontaneously hypertensive rats (SHR), and aged Wistar rats were used. Rats in each group were randomly subject to four experimental conditions: - Control: rats with ad libitum water intake, receiving vehicle (0.9% NaCl, i.p). - Cisplatin: rats with ad libitum water intake, treated with 2.5 mg/kg, i.p. cisplatin. - Water deprivation: rats deprived of water for 48 h, receiving vehicle (0.9% NaCl, i.p.). - Water deprivation + cisplatin: rats deprived of water for the 48 h prior to 2.5 mg/kg i.p. cisplatin administration. Tail vein blood and 24-hour urine samples were collected at basal time (B), after water deprivation (D0), and 4 days after cisplatin/vehicle administration (D4, day of maximum kidney damage). Haematocrit was measured and plasma osmolality, plasma creatinine (Jaffe reaction) and urea (Jung method) concentrations were determined. Urine samples were analysed for volume (urinary flow), osmolality, and proteinuria (Bradford method). Body fluid composition was measured by bioimpedance spectroscopy with an ImpediVETTM VetBIS1 device. Results All rats showed clear signs of dehydration after 48 hours of water deprivation: weight loss, increases in plasma osmolality and haematocrit (more pronouncedly in young Wistar and SHR than in aged animals), and reduced flow of a highly concentrated urine (i.e., with elevated osmolality). Bioimpedance analysis revealed a parallel loss of intracellular and extracellular water and net loss of total body water (explaining most of weight loss), with no changes in the liquid distribution between the intracellular and extracellular compartments. Dehydration was reverted after 4 days of rehydration in vehicle-treated rats, and no alteration of renal function was observed due to water deprivation. Administration of cisplatin in young, normohydrated Wistar rats showed minimal loss of renal function that was not worsened by water deprivation. Hypertensive rats showed completely normal renal function when cisplatin was administered under normohydration. Interestingly, however, dehydrated SHR did suffer a cisplatin-induced AKI (significant increases in plasma creatinine, plasma urea and proteinuria). In aged Wistar rats, cisplatin compromised renal function (as supported by increases in plasma creatinine and urea concentration), and this effect was significantly amplified by water deprivation. Conclusion This study suggests that dehydration in fully competent young rats is not in itself a significant risk factor for AKI, while the combination of dehydration and hypertension in young rats significantly increases the risk of AKI. Furthermore, ageing poses animals in a state of frailty that renders their kidneys unable to respond to stressors (such as a low dose of cisplatin). Frailty is boldly worsened in dehydrated aged animals. In perspective, models reproducing conditions of increased vulnerability to AKI provide useful tools to search for biomarkers pre-emptively predicting undesired health outcomes before exposure to stressors, and for developing preventing strategies. This study was supported by Project PI21/01226, funded by Instituto de Salud Carlos III (ISCIII) and co-funded by the European Union, and a grant from the Consejería de Educación, Junta de Castilla y León (IES160P20), Spain, co-funded by FEDER funds. Noelia Diaz-Morales is recipient of a Juan de la Cierva-Formación postdoctoral contract (FJC2020-043205-I) funded by MCIN/AEI/10.13039/501100011033 and European Union “NextGenerationEU/PRTR”.

MiR-328 - a novel diagnostic and predictive atrial fibrillation biomarker in spontaneously hypertensive rats

Abstract Funding Acknowledgements Type of funding sources: Public grant(s) – National budget only. Main funding source(s): Romanian Ministry of Education and Research, CNCS-UEFISCDI Purpose Atrial fibrillation (AF) is the most common cardiac arrhythmia and a major cause of morbidity and mortality worldwide. To date, there are no reliable biomarkers for the diagnosis and prediction of AF. Alterations in the atrial expression of various microRNAs contributes to the pathogenesis of AF, suggesting that certain miRNAs may emerge as promising AF diagnostic and/or predictive biomarkers. We aimed to evaluate miR-328 as a potential biomarker for the prediction and diagnosis of AF in an experimental model of spontaneous AF in rats. Method Male spontaneously hypertensive rats (SHR) and their normotensive counterparts (WKY) were divided into 6 groups: young (12 weeks, n = 8 each), adult (26 weeks, n = 8 each), and elderly (46 weeks, n = 10 each) SHR and WKY. All animals were implanted with ECG radiotelemetry devices, the ECG was continuously monitored for 72 hours, and the number of spontaneous AF episodes was assessed. At the end of the protocol, blood and left atrium tissue samples were taken and miR-328 levels were quantified. miR-328 levels were normalized with U6 snRNA and compared between groups. Correlations between atrial and blood miR-328 levels, and between mir-328 levels and arrhythmic load were also assessed. Results None of the WKY rats and none of the young SHRs presented arrhythmic episodes; two of the adult SHRs and all elderly SHRs had spontaneous AF episodes. Young SHR and WKY rats had similar atrial miR-328 levels (p = 0.67). At adult age, atrial miR-328 levels were significantly higher in SHRs than in WKYs (1.20 ± 0.02 vs. 1.07 ± 0.04; p = 0.01). Between-groups differences were even higher in the elderly rats (1.59 ± 0.01 vs. 1.03 ± 0.03; p< 0.01). Among the SHRs, atrial miR-328 levels increased progressively with age (p = 0.02) and were strongly correlated with the circulating blood levels of miR-328 (r = 0.71; p = 0.04). Moreover, among the elderly SHRs, atrial miR-328 levels correlated positively with the number of arrhythmic episodes (r = 0.72; p< 0.01). Conclusions The present study demonstrates a strong association between increased miR-328 expression levels and the presence and burden of AF in the hypertensive rats, suggesting that miR-328 could be a promising biomarker for the diagnosis of AF. Moreover, our results demonstrate that in this model the increase in miR-328 levels precedes the onset of AF, suggesting that dynamic evaluation of miR-328 could be used to predict AF, at least in a hypertensive context. Further studies will have to validate these results in clinical settings and to assess miR-328 manipulation as a potential prophylactic and/or therapeutic strategy in AF.

Analysis of salivary flow rate, biochemical composition, and redox status in orchiectomized spontaneously hypertensive rats

ObjectiveThis study aimed to analyze the salivary flow rate, biochemical composition, and redox status in orchiectomized spontaneously hypertensive rats (SHR) compared to normotensive Wistar rats. DesignThirty-two young adult male SHR and Wistar (3-months-old) rats were randomly distributed into four groups; either castrated bilaterally (ORX) or underwent fictitious surgery (SHAM) as Wistar-SHAM, Wistar-ORX, SHR-SHAM, and SHR-ORX. Two months beyond castration, pilocarpine-induced salivary secretion was collected from 5-month-old rats to analyze salivary flow rate, pH, buffer capacity, total protein, amylase, calcium, phosphate, sodium, potassium, chloride, thiobarbituric acid reactive substances (TBARs), carbonyl protein, nitrite, and total antioxidant capacity. ResultsThe salivary flow rate was higher in the Wistar-ORX compared to the Wistar-SHAM group, while remaining similar between the SHR-SHAM and SHR-ORX groups. ORX did not affect pH and salivary buffer capacity in both strains. However, salivary total protein and amylase were significantly reduced in the Wistar-ORX and SHR-ORX compared to the respective SHAM groups. In both ORX groups, salivary total antioxidant capacity and carbonylated protein were increased, while lipid oxidative damage (TBARs) and nitrite concentration were higher only in the Wistar-ORX than in the Wistar-SHAM group. In the Wistar-ORX and SHR-ORX, the salivary calcium, phosphate, and chloride were increased while no change was detected in the SHAM groups. Only salivary buffering capacity, calcium, and chloride in the SHR-ORX adjusted to values similar to Wistar-SHAM group. ConclusionHypertensive phenotype mitigated the orchiectomy-induced salivary dysfunction, since the disturbances were restricted to alterations in the salivary biochemical composition and redox state.

Egg white protein hydrolysate decreased blood pressure via the competing endogenous RNA regulatory networks in female spontaneously hypertensive rats.

Despite numerous studies having reported the effects and mechanisms of antihypertensive peptides including peptides derived from egg white proteins, the role of peptides in a female hypertensive animal model is unknown. On the other hand, the role of epigenetic modulation by peptide treatment has rarely been investigated. This study sought to investigate the effect of egg white protein hydrolysate (EWH) in female spontaneously hypertensive rats (SHRs) as well as to explore the underlying mechanisms from the perspectives of the transcriptome and the profiles of non-coding RNAs. Young (12-14-week-old) female SHRs were orally administered 250 mg per kg body weight (low-dose) or 1000 mg per kg body weight (high-dose) EWH daily for 10 weeks. The blood pressure of the rats was monitored weekly. The mRNA and non-coding RNAs (miRNA, lncRNA, and circRNA) in the aorta were profiled by the high-throughput RNA-seq technique. Differentially expressed (DE) RNAs in the aorta were identified for the construction of the competing endogenous RNA (ceRNA) networks and key molecules were validated by qRT-PCR. The treatment of the high-dose EWH showed a significant effect on reducing blood pressure in female SHRs. Bioinformatic analyses revealed 813, 90, 347 and 869 DE-mRNAs, DE-miRNAs, DE-lncRNAs and DE-circRNAs, respectively. The CNTN5-LncRNA-XR_001835895.1-miR-384-5p was identified as the central network which was validated in the aorta and circulation of female SHRs. The results from this study demonstrated that the treatment with EWH reduced blood pressure via regulating the ceRNA networks in female SHRs, which provided novel insights into the mechanisms of food protein-derived antihypertensive peptides.

Effect of Metabolic Adaptation by Voluntary Running Wheel Activity and Aldosterone Inhibition on Renal Function in Female Spontaneously Hypertensive Rats.

Metabolic effects of physical activity may be reno-protective in the context of hypertension, although exercise stresses kidneys. Aldosterone participates in renal disease in hypertension, but exercise affects the plasma concentration of aldosterone. This study was designed to evaluate whether physical activity and pharmacological treatment by aldosterone have additive effects on renal protection in hypertensive rats. Female spontaneously hypertensive rats (SHR) or normotensive Wistar rats performed voluntary running wheel activity alone or in combination with aldosterone blockade (spironolactone). The following groups were studied: young and pre-hypertensive SHR (n = 5 sedentary; n = 10 running wheels, mean body weight 129 g), 10-month-old Wistar rats (n = 6 sedentary; n = 6 running wheels, mean body weight 263 g), 10-month-old SHRs (n = 18 sedentary, mean body weight 224 g; n = 6 running wheels, mean body weight 272 g; n = 6 aldosterone, mean body weight 219 g; n = 6 aldosterone and running wheels, mean body weight 265 g). Another group of SHRs had free access to running wheels for 6 months and kept sedentary for the last 3 months (n = 6, mean body weight 240 g). Aldosterone was given for the last 4 months. SHRs from the running groups had free access to running wheels beginning at the age of 6 weeks. Renal function was analyzed by microalbuminuria (Alb/Cre), urinary secretion of kidney injury molecule-1 (uKim-1), and plasma blood urea nitrogen (BUN) concentration. Molecular adaptation of the kidney to hypertension and its modification by spironolactone and/or exercise were analyzed by real-time PCR, immunoblots, and histology. After six months of hypertension, rats had increased Alb/Cre and BUN but normal uKim-1. Voluntary free running activity normalized BUN but not Alb/Cre, whereas spironolactone reduced Alb/Cre but not BUN. Exercise constitutively increased renal expression of proprotein convertase subtilisin/kexin type 9 (PCSK9; mRNA and protein) and arginase-2 (mRNA). Spironolactone reduced these effects. uKim-1 increased in rats performing voluntary running wheel activity exercise irrespectively of blood pressure and aldosterone blockade. We observed independent but no additive effects of aldosterone blockade and physical activity on renal function and on molecules potentially affecting renal lipid metabolism.

How Do Young and Old Spontaneously Hypertensive Rats Respond to Antihypertensive Therapy? Comparative Studies on the Effects of Combined Captopril and Nifedipine Treatment.

Numerous studies on the effects of antihypertensive treatment in young spontaneously hypertensive rats (SHRs) have shown that early-onset therapy may effectively reduce their blood pressure (BP) even to normotensive values. In contrast, only a few studies investigated the effects of treatment started at an advanced age. These studies revealed that antihypertensive effects are lower in adult or even in senescent SHRs compared with young SHRs. Even more, prevention of cardiac sequelae of hypertension such as hypertrophy and fibrosis is less effective when treatment starts late in life. Because, in patients, combination therapies with calcium antagonists are favored, we studied the efficacy of a combination therapy with captopril and nifedipine in young and old SHRs. We directly compared the treatment effects on BP as well as on cardiac hypertrophy and remodeling between these two animal cohorts. With antihypertensive treatment, significantly lower BP values were achieved in young SHRs despite a shorter treatment period compared with old SHRs. Although treatment effects on cardiac hypertrophy were greater in old than in young SHRs, cardiac fibrosis was significantly attenuated only in young but not in old SHRs. The results emphasize the value of antihypertensive therapy and particularly accentuate the importance of an early-onset therapy. With respect to problems such as late diagnosis and poor therapy adherence, these results may have great importance for the treatment of human hypertension.

Sexual dimorphism in renal heme oxygenase-1 and arachidonic acid metabolizing enzymes in spontaneously hypertensive rats versus normotensive Wistar Kyoto rats

Numerous studies have demonstrated a sexual dimorphism in blood pressure (BP) control in spontaneously hypertensive rats (SHR), however the mechanisms remain to be further elucidated. Based on the established role of arachidonic acid metabolites and heme oxygenase (HO) in BP control, we hypothesize that higher BP in male SHR is associated with differential expression in renal HO and arachidonic acid metabolizing enzymes vs. female SHR. Higher BP in male SHR coincided with significant increases in renal cortical superoxide production and thiobarbituric acid reactive substances (TBARs) levels as measures of oxidative stress compared to normotensive female WKY and female SHR. The elevations in BP and oxidative stress in male SHR were also associated with a decrease in cortical heme oxygenase-1 (HO-1) expression when compared to normotensive female WKY. Although there was no sex or strain differences in cortical expression of the epoxyeicosatrienoic acids (EETs) producing enzyme, cytochrome P450 epoxygenase (CYP2C23), in male and female SHR and WKY, SHR had greater expression of the EETs metabolizing enzyme, soluble epoxide hydrolase (sEH) vs. WKY. Cortical expression of the 20–hydroxyeicosatetraenoic acid (20-HETE) producing enzyme, cytochrome P450 hydroxylase (CYP4A), was less in female WKY and SHR compared to strain-matched males and cortical 20-HETE levels were also less in female SHR vs. male SHR. Cortical cyclooxygenase-2 (COX-2) expression was significantly greater in female SHR and WKY vs. males and cortical prostaglandin E2 (PGE2) levels in female SHR was significantly greater than male WKY. In conclusion, our data suggest that sex differences in renal oxidative stress, HO-1 and arachidonic acid metabolizing enzymes could contribute to sexual dimorphism in hypertension in young SHR.

Treatment of male and female spontaneously hypertensive rats with TNF-\u03b1 inhibitor etanercept increases markers of renal injury independent of an effect on blood pressure

Hypertension remains the leading risk factor for cardiovascular disease. Young females tend to be protected from hypertension compared with age-matched males. Although it has become increasingly clear that the immune system plays a key role in the development of hypertension in both sexes, few studies have examined how cytokines mediate hypertension in males versus females. We previously published that there are sex differences in the levels of the cytokine tumor necrosis factor (TNF)-α in spontaneously hypertensive rats (SHR). The goal of this study was to test the hypothesis that TNF-α inhibition with etanercept will lower BP in male and female SHR. However, as male SHR have a more pro-inflammatory status than female SHR, we further hypothesize that males will have a greater decrease in BP with TNF-α inhibition than females. Young adult male and female SHR were administered increasing doses of the TNF-α inhibitor etanercept or vehicle twice weekly for 31 days and BP was continuously measured via telemetry. Following treatment, kidneys and urine were collected and analyzed for markers of inflammation and injury. Despite significantly decreasing renal TNF-α levels, renal phospho-NFκB and urinary MCP-1 excretion, etanercept did not alter BP in either male or female SHR. Interestingly, treatment with etanercept increased urinary excretion of protein, creatinine and KIM-1 in both sexes. These results indicate that TNF-α does not contribute to sex differences in BP in SHR but may be vital in the maintenance of renal health.

Epoxyeicosatrienoic Acid Analog and 20-HETE Antagonist Combination Prevent Hypertension Development in Spontaneously Hypertensive Rats.

Numerous studies indicate a significant role for cytochrome P-450-dependent arachidonic acid metabolites in blood pressure regulation, vascular tone, and control of renal function. Epoxyeicosatrienoic acids (EETs) exhibit a spectrum of beneficial effects, such as vasodilatory activity and anti-inflammatory, anti-fibrotic, and anti-apoptotic properties. 20-Hydroxyeicosatetraenoic acid (20-HETE) is a potent vasoconstrictor that inhibits sodium reabsorption in the kidney. In the present study, the efficiency of EET-A (a stable analog of 14,15-EET) alone and combined with AAA, a novel receptor antagonist of 20-HETE, was tested in spontaneously hypertensive rats (SHR). Adult SHR (16 weeks old) were treated with two doses of EET-A (10 or 40 mg/kg/day). In the following experiments, we also tested selected substances in the prevention of hypertension development in young SHR (6 weeks old). Young rats were treated with EET-A or the combination of EET-A and AAA (both at 10 mg/kg/day). The substances were administered in drinking water for 4 weeks. Blood pressure was measured by telemetry. Once-a-week observation in metabolic cages was performed; urine, blood, and tissue samples were collected for further analysis. The combined treatment with AAA + EET-A exhibited antihypertensive efficiency in young SHR, which remained normotensive until the end of the observation in comparison to a control group (systolic blood pressure, 134 ± 2 versus 156 ± 5 mmHg, respectively; p < 0.05). Moreover the combined treatment also increased the nitric oxide metabolite excretion. Considering the beneficial impact of the combined treatment with EET-A and AAA in young rats and our previous positive results in adult SHR, we suggest that it is a promising therapeutic strategy not only for the treatment but also for the prevention of hypertension.

Abstract P213: Furosemide-mediated Suppression Of P 38 MAPK Signaling Abrogates Cholinergic-mediated Salt Sensitive Hypertension In Young Spontaneously Hypertensive Rats

Hypertension is a major cause of premature death worldwide. Inflammatory macrophages play a key role in the pathogenesis of hypertension. Our lab has observed that nicotinic acetylcholine receptors (nAChR) activation potentiates salt-sensitive hypertension in Spontaneously Hypertensive Rats (SHR). Cholinergic receptor activation causes infiltration of CD68 + macrophages into the renal cortex/medulla junction and results in enhanced expression of renal sodium-potassium chloride cotransporter (NKCC2). To determine whether or not NKCC2 plays a role in the activation of inflammatory pathway furosemide, a selective pharmacologic blocker of NKCC2, was used to block NKCC2 expression in SHR in current study. Young pre-hypertensive SHR, receiving nicotine infusion (15mg/kg/day), were fed HSD (4% NaCl) and treated with either saline or furosemide (20mg/kg/day) intraperitoneal injections. There was a 16% increase (158.5±7.3 mmHg to 183.6±7.0 mmHg) in the systolic blood pressure in SHR that received saline injections, compared to no increase in SHR that received furosemide (155±6.0 mmHg to 148±3.6 mmHg) (p&lt;0.0016). There was over a two-fold increase in fractional excretion of sodium (FENa, p=0.03) in the furosemide group, compared to saline controls. Although furosemide injections did not alter renal NKCC2 expression, there was nearly 50% reduction in phosphorylated p38 MAPK (p=0.0002) and a 25% decrease in renal infiltration of inflammatory CD68 + macrophages in SHR receiving furosemide injections, compared to saline controls. Interestingly, there was also a significant reduction in urinary albumin in response to furosemide injections (14.7±5.7 control vs 3.3±0.9 furosemide, p=0.027). Based on these findings, we conclude that furosemide results in cessation of the development of hypertension via interruption of a p38-MAPK dependent inflammatory pathway.

Abstract P216: Cholinergic-mediated Salt-Sensitive Hypertension In A Genetic Model Of Essential Hypertension

Inflammation plays a key role in the pathogenesis of hypertension. High dietary sodium intake has been associated with inflammation in patients suffering from high blood pressure &amp; atherosclerosis. Our lab has shown that development of hypertension in a genetic model of human essential hypertension, the Spontaneously Hypertensive Rat (SHR), is dependent on nicotinic acetylcholine receptor (nAChR) mediated renal inflammation, which leads to enhanced expression of the renal sodium-potassium chloride cotransporter (NKCC2). Based on these findings we hypothesized that sodium sensitivity may play a role in the development of cholinergic hypertension. To investigate this hypothesis, we infused young pre-hypertensive SHR and Wistar Kyoto (WKY, normotensive control) rats with either saline or nicotine (15mg/kg/day) and placed them on either low-salt (LSD, 0.01% NaCl) or high-salt (HSD, 4% NaCl) diet for 2 wks. There was no change in systolic blood pressure (SBP) in saline or nicotine infused WKY with either LSD or HSD. Similarly, there was no difference between the SBP of saline infused SHR with either LSD or HSD. Interestingly, nicotine infusion did not change SBP in LSD fed SHR, but did result in ~20% increase in SBP in HSD fed SHR (122.0 ± 0.7 vs. 146.2 ± 0.2mmHg, p= 0.01). HSD alone induced a 2.25-fold increase in renal NKCC2 expression compared to LSD in saline infused SHR (p=0.006). In comparison to saline, nicotine infusion further induced a 58% increase in renal NKCC2 expression in SHR fed HSD (p=0.004). Renal NKCC2 expression did not change in WKY with either diet or nicotine infusion. Nicotine infused SHR on a HSD also manifested a 16% sodium retention compared to saline infused SHR (2.1 ± 0.1 mM/gBW vs. 2.5 ± 0.1 mM/gBW, p = 0.017). Nicotine infused SHR had 1.7 times more renal IL-6 than saline-infused SHR (1442. ± 212 vs 826 ± 103 pg/25mg total protein, p=0.04) and 9 times more renal IL-6 than nicotine-infused WKY controls (1442. ± 212 vs 148. ± 71 pg/25mg total protein, p=0.0041) under HSD conditions. Taken together with our previously published data, we conclude that NKCC2 likely plays a role in cholinergic-mediated renal inflammation and hypertension.

MP08-14 AGING CAUSES DETRUSOR UNDERACTIVITY IN SPONTANEOUSLY HYPERTENSIVE RATS

You have accessJournal of UrologyBladder & Urethra: Anatomy, Physiology & Pharmacology (MP08)1 Sep 2021MP08-14 AGING CAUSES DETRUSOR UNDERACTIVITY IN SPONTANEOUSLY HYPERTENSIVE RATS Shogo Shimizu, Yoshiki Nagao, Suo Zou, Takaaki Aratake, Takahiro Shimizu, Youichirou Higashi, and Motoaki Saito Shogo ShimizuShogo Shimizu More articles by this author , Yoshiki NagaoYoshiki Nagao More articles by this author , Suo ZouSuo Zou More articles by this author , Takaaki AratakeTakaaki Aratake More articles by this author , Takahiro ShimizuTakahiro Shimizu More articles by this author , Youichirou HigashiYouichirou Higashi More articles by this author , and Motoaki SaitoMotoaki Saito More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000001981.14AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Although aging or severe bladder ischemia could be involved in progression of detrusor underactivity (DU), the pathophysiological mechanism of DU is not well clarified. Hypertension is one of most common chronic disease among elderly, affecting bladder function and blood flow in lower urinary tract. In this study, we investigated involvement of aging on bladder function in spontaneously hypertensive rats (SHRs). METHODS: Male SHRs and Wistar Kyoto rats (WKYs) at the age of 18, 36, 54 or 72 weeks (W) were used (n = 7, each group). WKYs were used as normotensive controls. At each ages, metabolic cage study, measuring blood pressure, and single and continuous cystometry were performed. Bladder blood flow (BBF) was measured by hydrogen clearance method. And, bladder weight was measured and bladder tissue was stained with hematoxylin and eosin staining. RESULTS: SHRs at 72 W showed significant increases in mean blood pressure, bladder weight/body weight ratio (BBR) and urine output for 24 h, and decrease in BBF compared to SHRs at 18 W. SHRs at 72 or 54 W showed significant increases in single voided volume (SVV), post residual urine volume (RV) and bladder capacity (BC), and decrease in voiding efficacy (VE) compared to SHRs at 18 W. SHRs at 54 or 72 W also showed significantly prolonged intercontraction interval (ICI) compared to SHRs at 18 W. Detrusor thickness in SHRs at 72 W were significantly larger than that in SHRs at 18 W. However, there were no significant differences on mean blood pressure, BBR, voiding parameters and detrusor thickness in WKYs among each age. CONCLUSIONS: Aged SHRs showed that significant increases in blood pressure, SVV, RV and urine output, prolonged ICI and decreases in VE and BBF compared to young SHRs. Aging related severe hypertension could cause DU via severe bladder ischemia and polyuria. Aged SHR might be one of useful animal model to clarify the pathophysiological mechanisms of DU. Source of Funding: This study was supported by JSPS KAKENHI [Grant no. 17K16797 and 19K09673 (SS)] © 2021 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 206Issue Supplement 3September 2021Page: e155-e155 Advertisement Copyright & Permissions© 2021 by American Urological Association Education and Research, Inc.MetricsAuthor Information Shogo Shimizu More articles by this author Yoshiki Nagao More articles by this author Suo Zou More articles by this author Takaaki Aratake More articles by this author Takahiro Shimizu More articles by this author Youichirou Higashi More articles by this author Motoaki Saito More articles by this author Expand All Advertisement Loading ...

Cardiac dysfunction in spontaneously hypertensive old rats is associated with a significant decrease of SUR2 expression.

ATP-sensitive potassium (KATP) channels are participants of mechanisms of pathological myocardial remodeling containment. The aim of our work was to find the association of changes in the expression of Kir6.1, Kir6.2, SUR1, and SUR2 subunits of KATP channels with changes in heart function and structure during aging under conditions of the constant increase of vascular pressure. The experiments were carried out on young and old spontaneously hypertensive rats (SHR) and Wistar rats. The expression levels of KATP channels subunits were determined using reverse transcription and quantitative PCR. It is shown that the mRNA expression level of Kir6.1 in young SHR rats is significantly lower (6.3-fold, p = 0.035) than that of young Wistar rats that may be one of the causes of arterial hypertension in SHR. At the same time, mRNA expression of both Kir6.1 and Kir6.2 in old SHR rats was significantly higher (6.8-fold, p = 0.003, and 5.9-fold, p = 0.006, respectively) than in young hypertensive animals. In both groups of old animals, SUR2 expression was significantly reduced compared to young animals, in Wistar rats at 3.87-fold (p = 0.028) and in SHR rats at 48.2-fold (p = 0.033). Changes in SUR1 expression were not significant. Thus, significant changes in the cardiovascular system, including impaired function and structure of the heart in old SHR rats, were associated with a significant decrease in SUR2 expression that may be one of the mechanisms of heart failure decompensation. Therefore, it can be assumed that increased expression of SUR2 may be one of the protective mechanisms against pathological myocardial remodeling.

Geometric Features of the Pial Arteriolar Networks in Spontaneous Hypertensive Rats: A Crucial Aspect Underlying the Blood Flow Regulation.

BackgroundSeveral studies indicate that hypertension causes major changes in the structure of the vessel wall by affecting the regulation of blood supply to the tissues. Recently, it has been observed that capillary blood flow is also considerably influenced by the structural arrangement of the microvascular networks that undergo rarefaction (reduction of the perfused vessel number). Therefore, this study aimed to assess the geometric arrangements of the pial arteriolar networks and the arteriolar rhythmic diameter changes in spontaneously hypertensive rats (SHRs).MethodsFluorescence microscopy was utilized to observe in vivo the pial microcirculation through a closed cranial window. Pial arterioles were classified according to Strahler’s method. The arteriolar rhythmic diameter changes were evaluated by a generalization short-time Fourier transform.ResultYoung SHRs showed four orders of vessels while the adult ones only three orders. The diameter, length, and branching number obeyed Horton’s law; therefore, the vessels were distributed in a fractal manner. Larger arterioles showed more asymmetrical branches than did the smaller ones in young SHRs, while in adult SHRs smaller vessels presented asymmetrical branchings. In adult SHRs, there was a significant reduction in the cross-sectional area compared with the young SHRs: this implies an increase in peripheral resistance. Young and adult age-matched normotensive rats did not show significant alterations in the geometric arteriolar arrangement with advancing age, both had four orders of arteriolar vessels, and the peripheral resistance did not change significantly. Conversely, the frequency components evaluated in arteriolar rhythmic diameter changes of young and adult SHRs showed significant differences because of a reduction in the frequency components related to endothelial activity detected in adult SHRs.ConclusionIn conclusion, hypertension progressively causes changes in the microarchitecture of the arteriolar networks with a smaller number of vessels and consequent reduced conductivity, characteristic of rarefaction. This was accompanied by a reduction in the formation and release of independent and dependent – endothelial nitric oxide components regulating arterial vasomotion.

CRISPR/Cas9 Mediated Deletion of the Angiotensinogen Gene Reduces Hypertension: A Potential for Cure?

[Figure: see text].

Morphofunctional state of the lungs respiratory part in normotensive and hypertensive rats after combined exposure to intermittent hypoxia and melatonin

Aim. The purpose of this work was to study and compare the combined effect of intermittent normobaric hypoxia (INH) and melatonin on the morphological and biochemical indices of the lungs respiratory part in Wistar and spontaneous hypertensive rats (SHR). Material and methods. The studies were conducted on 48 young male rats Wistar and SHR lines. The experimental rats were daily exposed to hypoxic gas mixture (12 % oxygen in nitrogen) in intermittent mode: 15 min deoxygenation / 15 min reoxygenation for 2 hours. Animals also received oral melatonin at a dose of 5 mg /1 kg of body weight once daily.Results. The sizes of alveolus were reduced, but their number and placement density were increased in SHR rats after a combined effect of INH and melatonin. The results showed a significant decrease of the interalveolar septum thickness and a tendency to decrease of the hydroxyproline concentration in the lung tissue of the experimental rats, regardless of their line.Conclusions. The 28-day combined effect of INH and melatonin increase the total alveolar surface area and reduce the amount of connective tissue in the lungs of experimental rats. These changes can improve the efficiency of the intrapulmonary gas exchange processes.

Возрастные изменения механизмов эндотелий-зависимой дилатации пиальных артериальных сосудов у крыс SHR

The study investigated the role of age-related changes in the contribution of NO and IKCa-channels to acetylcholine-induced dilation of pial arteries in spontaneously hypertensive rats (SHR). We used intravital microphotography (×470) to evaluate the responses of pial arteries to acetylcholine chloride (ACh, 10−7 M, 5 min) with and without blockade of NO (L-NAME, 10−3 M) and IKCa-channels (klotrimazole,10−5 M) in SHR rats aged 4 and 18 months. The number and degree of vascular dilatation was measured from the erythrocyte flow width in three individual groups of arteries: small (less than 20 μm in diameter), medium (20–40 μm) and large (more than 40 μm). It was found that in young SHR rats, NO plays a significant role in the ACh-mediated dilation of small and large-diameter vessels. The role of IKCa-channels in endothelium-dependent dilatation mainly manifests itself in the group of small vessels and decreases with an increase in artery diameter. Aging, accompanied by hypertension, reduces the role of IKCa-channels and increases the contribution of NO to the dilatation of pial arterial vessels of all sizes.

ВОЗРАСТ-ЗАВИСИМЫЕ ИЗМЕНЕНИЯ РОЛИ КАЛИЕВЫХ КАНАЛОВ В ДИЛАТАЦИИ ПИАЛЬНЫХ АРТЕРИЙ НА АЦЕТИЛХОЛИН У КРЫС WKY И SHR

Age-related changes in the contribution of BKCa-channels and NO to acetylcholine-induced dilation of pial arteries were studied in normotensive (Wistar-Kyoto) and spontaneously hypertensive rats (SHRs). Using intravital microphotography (×470), responses of pial arteries to acetylcholine chloride (ACh, 10−7 M, 5 min) with and without blockade of BKCa-channels by tetraethylammonium chloride (TEA, 2 mМ) and nitric oxide (NO) synthesis by L-NAME (10−3 M) were comparatively evaluated in WKY rats and SHRs aged 4 and 20 months. The evaluation criteria were the number and degree of arterial dilations arising in response to ACh after the application of inhibitors, with the latter criterion being evaluated, in turn, by measuring the erythrocyte flow width separately in three groups of arteries: small ( 40 µm). It was established that in WKY rats aging leads to diminish the role of BKCa-channels in ACh-induced dilation of all pial arteries, no matter the caliber. Similar processes were observed in the pial vascular bed of young SHRs. It appears that changes in the role of BKCa-channels in vascular dilatatory responses in aging WKY rats and young SHRs are largely associated with the impairment of NO synthesis and changes in the role of NO-mediated mechanisms of vasodilation. Aging in SHRs is accompanied by increasing the contribution of the NO-dependent mechanism to the regulation of ACh-induced dilatatory responses of small-caliber arteries.

Acute Effects of Ibrutinib on Ventricular Arrhythmia in Spontaneously Hypertensive Rats

BackgroundThe Bruton's Tyrosine Kinase Inhibitor ibrutinib is associated with ventricular arrhythmia (VA) and sudden death. However, the pro-arrhythmic electrophysiological dysregulation that results from ibrutinib with age and cardiovascular disease is unknown. ObjectivesThis study sought to investigate the acute effects of ibrutinib on left ventricular (LV) VA vulnerability, cytosolic calcium dynamics, and membrane electrophysiology in old and young spontaneous hypertensive rats (SHRs). MethodsLangendorff-perfused hearts of young (10 to 14 weeks) and old (10 to 14 months) SHRs were treated with ibrutinib (0.1 μmol/l) or vehicle for 30 min. Simultaneously, LV epicardial action potential and cytosolic calcium transients were optically mapped following an incremental pacing protocol. Calcium and action potential dynamics parameters were analyzed. VA vulnerability was assessed by electrically inducing ventricular fibrillations (VFs) in each heart. Western blot analysis was performed on LV tissues. ResultsIbrutinib treatment resulted in higher vulnerability to VF in old SHR hearts (27.5 ± 7.5% vs. 5.7 ± 3.7%; p = 0.026) but not in young SHR hearts (8.0 ± 4.9% vs. 0%; p = 0.193). In old SHR hearts, following ibrutinib treatment, action potential duration (APD) alternans (p = 0.008) and APD alternans spatial discordance (p = 0.027) were more prominent. Moreover, calcium transient duration 50 was longer (p = 0.032), calcium amplitude alternans ratio was significantly lower (p = 0.001), and time-to-peak of calcium amplitude was shorter (p = 0.037). In young SHR hearts, there were no differences in calcium and APD dynamics. ConclusionsIbrutinib-induced VA is associated with old age in SHR. Acute dysregulation of calcium and repolarization dynamics play important roles in ibrutinib-induced VF.

Hypoxia-mediated regulation of mitochondrial transcription factors in renal epithelial cells: implications for hypertensive renal physiology.

Kidneys have a high resting metabolic rate and low partial pressure of oxygen due to enhanced mitochondrial oxygen consumption and ATP production needed for active solute transport. Heightened mitochondrial activity leads to progressively increasing hypoxia from the renal cortex to the renal medulla. Renal hypoxia is prominent in hypertensive rats due to increased sodium reabsorption within the nephrons, which demands higher energy production by oxidative phosphorylation (OXPHOS). Consequently, spontaneously hypertensive rats (SHR) display greater oxygen deficiency (hypoxia) than normotensive Wistar Kyoto rats (WKY). Here, we sought to investigate the expression of key proteins for mitochondrial biogenesis in SHR and WKY, and study the regulation of mitochondrial transcription factors (mtTFs) under in vitro hypoxic conditions in renal epithelial cells. We report that renal expressions of hypoxia-inducible factor-1-alpha (HIF-1α), peroxisome proliferator-activated receptor-gamma coactivator-1-alpha (PGC-1α), mtTFs, and OXPHOS proteins are elevated in SHR compared to WKY. In addition, our experiments in cultured kidney cells demonstrate that acute hypoxia augments the expression of these genes. Furthermore, we show that the transcripts of HIF-1α and mtTFs are positively correlated in various human tissues. We reveal, for the first time to our knowledge, that HIF-1α transactivates mtTF genes by direct interaction with their promoters in rat kidney epithelial cells (NRK-52E) under acute hypoxia. Concomitant increases in the mitochondrial DNA and RNA, and OXPHOS proteins are observed. Taken together, this study suggests that hypoxia within the renal epithelial cells may enhance mitochondrial function to meet the energy demand in proximal tubular cells during prehypertensive stages in kidneys of young SHR.