Overactivity Of Renin-angiotensin System Research Articles

Angiotensin II Alters Mitochondrial Membrane Potential and Lipid Metabolism in Rat Colonic Epithelial Cells.

An over-active renin-angiotensin system (RAS) is characterized by elevated angiotensin II (Ang II). While Ang II can promote metabolic and mitochondrial dysfunction in tissues, little is known about its role in the gastrointestinal system (GI). Here, we treated rat primary colonic epithelial cells with Ang II (1-5000 nM) to better define their role in the GI. We hypothesized that Ang II would negatively affect mitochondrial bioenergetics as these organelles express Ang II receptors. Ang II increased cellular ATP production but reduced the mitochondrial membrane potential (MMP) of colonocytes. However, cells maintained mitochondrial oxidative phosphorylation and glycolysis with treatment, reflecting metabolic compensation with impaired MMP. To determine whether lipid dysregulation was evident, untargeted lipidomics were conducted. A total of 1949 lipids were detected in colonocytes spanning 55 distinct (sub)classes. Ang II (1 nM) altered the abundance of some sphingosines [So(d16:1)], ceramides [Cer-AP(t18:0/24:0)], and phosphatidylcholines [OxPC(16:0_20:5(2O)], while 100 nM Ang II altered some triglycerides and phosphatidylserines [PS(19:0_22:1). Ang II did not alter the relative expression of several enzymes in lipid metabolism; however, the expression of pyruvate dehydrogenase kinase 2 (PDK2) was increased, and PDK2 can be protective against dyslipidemia. This study is the first to investigate the role of Ang II in colonic epithelial cell metabolism.

Renal autonomic dynamics in hypertension: how can we evaluate sympathetic activity for renal denervation?

This review explores the various pathophysiological factors influencing antihypertensive effects, involving the regulation of vascular resistance, plasma volume, cardiac function, and the autonomic nervous system, emphasizing the interconnected processes regulating blood pressure (BP). The kidney's pivotal role in BP control and its potential contribution to hypertension is complicated but important to understand the effective mechanisms of renal denervation (RDN), which may be a promising treatment for resistant hypertension. Excessive stimulation of the sympathetic nervous system or the renin-angiotensin-aldosterone system (RAAS) can elevate BP through various physiological changes, contributing to chronic hypertension. Renal sympathetic efferent nerve activation leads to elevated norepinephrine levels and subsequent cascading effects on vasoconstriction, renin release, and sodium reabsorption. RDN reduces BP in resistant hypertension by potentially disrupting sensory afferent nerves, decreasing feedback activation to the central nervous system, and reducing efferent sympathetic nerve activity in the heart and other structures. RDN may also modulate central sympathetic outflow and inhibit renal renin-angiotensin system overactivation. While evidence for RDN efficacy in hypertension is increasing, accurate patient selection becomes crucial, considering complex interactions that vary among patients. This review also discusses methods to evaluate autonomic nerve activity from the golden standard to new potential examination for finding out optimization in stimulation parameters or rigorous patient selection based on appropriate biomarkers.

Importance of the (Pro)renin Receptor in Activating the Renin-Angiotensin System During Normotensive and Preeclamptic Pregnancies.

For a healthy pregnancy to occur, a controlled interplay between the maternal circulating renin-angiotensin-aldosterone system (RAAS), placental renin-angiotensin system (RAS) and intrarenal renin-angiotensin system (iRAS) is necessary. Functionally, both the RAAS and iRAS interact to maintain blood pressure and cardiac output, as well as fluid and electrolyte balance. The placental RAS is important for placental development while also influencing the maternal circulating RAAS and iRAS. This narrative review concentrates on the (pro)renin receptor ((P)RR) and its soluble form (s(P)RR) in the context of the hypertensive pregnancy pathology, preeclampsia. The (P)RR and the s(P)RR have become of particular interest as not only can they activate prorenin and renin, thus influencing levels of angiotensin II (Ang II), but s(P)RR has now been shown to directly interact with and stimulate the Angiotensin II type 1 receptor (AT1R). Levels of both placental (P)RR and maternal circulating s(P)RR are elevated in patients with preeclampsia. Furthermore, s(P)RR has been shown to increase blood pressure in non-pregnant and pregnant rats and mice. In preeclamptic pregnancies, which are characterised by maternal hypertension and impaired placental development and function, we propose that there is enhanced secretion of s(P)RR from the placenta into the maternal circulation. Due to its ability to both activate prorenin and act as an AT1R agonist, excess maternal circulating s(P)RR can act on both the maternal vasculature, and the kidney, leading to RAS over-activation. This results in dysregulation of the maternal circulating RAAS and overactivation of the iRAS, contributing to maternal hypertension, renal damage, and secondary changes to neurohumoral regulation of fluid and electrolyte balance, ultimately contributing to the pathophysiology of preeclampsia.

Novel Insights in the Hypertension Treatment & Type 2 Diabetics Induced by Angiotensin Receptor Blockers: MD Simulation Studies & Molecular Docking of Some Promising Natural Therapies.

Angiotensin receptor blockers (ARBs) are commonly used to treat hypertension that target the hormonal system (renin-angiotensin system (RAS)), which regulates various physiological functions in the body. ARBs work by blocking the binding of angiotensin II to its receptor, thereby preventing a rise in blood pressure. These drugs not only normalize the overactivation of RAS but also provide protective effects against cardiovascular, renal, and type 2 diabetic patients. Inappropriate RAS activity has been linked to insulin resistance of type 2 diabetes. Olmesartan, as an ARB, was found to have a beneficial role in reducing postprandial glucose levels in type 2 diabetes. However, ARBs can cause side effects, prompting a search for new compounds that have fewer adverse effects. This study explores the potential of natural metabolites, specifically eugenol, gallic acid, myricetin, p-cymene, quercetin, and kaempferol, as ARB inhibitors compared to the current standard, olmesartan. Using in silico studies, the binding affinity of these natural substances to the ARB receptor was evaluated. The results showed that myricetin and kaempferol had affinities higher than those of olmesartan, suggesting that they could serve as promising ARB inhibitors for hypertension treatment. These natural compounds could provide an alternative approach to conventional antihypertensive drugs, which may have fewer side effects. However, more research is needed to validate the efficacy and safety of these natural compounds as antihypertensive drugs. Further in vitro and in vivo studies are needed to confirm their effectiveness and safety. This study provides a promising starting point for future investigations into the potential of natural metabolites as alternative treatments for hypertension. The findings also highlight the importance of exploring natural alternative treatments for hypertension and the protective effects of ARBs on early stage type-2 diabetics.

Sex and tissue-specific downregulation of miR410-3p in a mouse model of cardiometabolic diseases

The brain Renin-Angiotensin System (RAS) plays a pivotal role in controlling cardiometabolic homeostasis, fluid balance, and other functions including inflammation. Previous studies have shown that more than a thousand microRNA (miRNA) families regulate RAS genes expression, and their dysregulation has been associated with HTN, cardiac ischemia, and aging. Recently, we observed that hypercaloric diet (HCD) programming resulted in differentially expressed miRNA in the hypothalamus of only the male progeny, including miR-410-3p which is thought to target angiotensin-II type 1 receptors (AT1R). Here, we aimed to investigate whether miR-410-3p regulate AT1R expression in a mouse model of cardiometabolic disease (CMD) exhibiting hypertension, diabetes and obesity. C57BI/6J mice (3-week-old, both sexes, n=10) were fed a HCD (D12492. 60 kcal% Fat, Research Diets, USA) while weight and fasting blood glucose levels were measured weekly to ensure the acquisition of metabolic syndrome. At 12 weeks of age, the mice were infused subcutaneously (osmotic pump model 1004, Alzet) with Ang-II (600 ng/kg/min/4 weeks). Gene expression of miR-410-3p (QRT-PCR) was reduced in the hypothalamus of CMD male mice (0.5 ±0.1 vs.1.0 ±0.1, p<0.05) but increased in females (1.6 ±0.4 vs.0.7 ±0.2, p<0.001), while both sexes showed a reduction in the heart (Males: 0.2 ±0.0 vs.1.7 ±0.3; Females: 0.23 ±0.0 vs. 3.2 ±0.3, p<0.01). AT1R protein expression (Capillary Western) was increased in the hypothalamus of CMD male (1.5 ±0.1 vs.1.0 ±0.2, p<0.05) mice but not in females (1.0 ±0.1 vs.1.2 ±0.1, P>0.05) while both sexes exhibited elevated levels in the heart (Males: 2.0 ±0.1 vs. 1.0 ±0.1; Females: 2.3 ±0.1 vs. 1.3 ±0.2, p<0.05). Binding specificity (luciferase assay) was further assessed between miR-410-3p and the 3’UTR of AT1R. Binding was decreased in presence of miR410-3p mimic (0.9 ±0.0 vs. 2.1 ±0.0; p<0.001) and enhanced by a miR410-3p inhibitor (2.5 ±0.1 vs. 2.1 ±0.0; p<0.01), confirming that AT1R is a direct target of miR410-3p. Together, our data suggest that in CMD downregulation of miR410-3p is associated with the upregulation of AT1R both in the hypothalamus and heart, notably in males. This positions miR410-3p as a possible new therapeutic strategy to control RAS overactivity in CMD. This work was supported in part by a research grant from the National Institutes of Health (HL163588). This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Upregulation of miR-361-3p Is Associated With Impaired AT2 Receptor Expression in Cardiometabolic Diseases

Cardiometabolic homeostasis has been shown to be regulated by the brain Renin-Angiotensin System (RAS), including via epigenetic mechanisms. MicroRNA (miRNA) regulation is one of the most common mechanisms controlling the expression of RAS genes and has previously been implicated in hypertension, cardiac ischemia, and aging. Recently, we observed that hypercaloric diet (HCD) programming resulted in differentially expressed miRNA in the hypothalamus of only the male progeny, including miR-361-3p which is thought to target angiotensin-II type 2 receptors (AT2R). Here, we aimed to investigate whether miR-361-3p regulate AT2R expression in a mouse model of cardiometabolic disease (CMD) exhibiting hypertension, diabetes and obesity. C57BI/6J mice (3-week-old, both sexes, n=10) were fed a HCD (D12492. 60 kcal% Fat, Research Diets, USA) while weight and fasting blood glucose levels were measured weekly to ensure the acquisition of metabolic syndrome. At 12 weeks of age, the mice were infused subcutaneously (osmotic pump model 1004, Alzet) with Ang-II (600 ng/kg/min/4 weeks). BP, food intake and glycemia were recorded weekly while intraperitoneal glucose (GTT), insulin (ITT) tolerance tests and body composition (EchoMRI) were assessed at the end of the study. Gene expression of miR-361-3p (QRT-PCR) was increased in the hypothalamus of CMD male mice (2.0 ±0.5 vs.1.2 ±0.2, p<0.05) but not in females (2.7 ±0.8 vs.2.5 ±0.4, P>0.05). However, both sexes showed an increase in the heart (Males: 2.0 ±0.5 vs.1.0 ±0.3; Females: 1.6 ±0.4 vs. 0.8 ±0.2, p<0.05) which was associated with decreased AT2R cardiac expression (Capillary Western) in CMD male (0.9 ±0.0 vs.1.0 ±0.0, p<0.05) and females (0.1 ±0.0 vs.0.6 ±0.1, p<0.001). Together, our data suggest that in CMD, upregulation of miR-361-3p is associated with the decrease of AT2R in the heart, possibly contributing to impaired cardiac function in this model. This positions inhibitors of miR-361-3p as a possible new therapeutic strategy to control RAS overactivity in CMD. This work was supported in part by a research grant from the National Institutes of Health (HL163588). This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Risk assessment of di-(2-ethylhexyl) phthalate plus a high-fat diet in the rat kidney

Di-(2-ethylhexyl) phthalate (DEHP) is used in food packaging and medical products, resulting in unintentional human ingestion. Concurrently, a high fat (HF)-oriented diet is very popular for many people worldwide. Here we investigated the regulatory role of renin-angiotensin system in kidney dysfunction. The HF diet-fed rats were fed a HF diet plus different doses of 10, 100 and 300 mg/kg bw DEHP for 35 days by gavage. Serum lipid contents and protein activities of renin-angiotensin system (RAS) components were measured using biochemical methods and ELISA; key metabolic genes and proteins of the kidney were determined by RT-PCR and western blotting techniques. Kidney morphology was examined through hematoxylin-eosin (HE) staining. Kidney weight to body weight ratio and mRNA expression of kidney damage markers were significantly increased following DEHP plus HF administration, which corresponded histologically with glomerular basement membrane thickening and glomerular swelling. Furthermore, DEHP treatment plus a HF diet induced dyslipidemia, accompanied by increased levels of renin-angiotensin system components, such as ACE protein, MAS-1 protein, serum renin and aldosterone. In addition, mRNA expression levels of pro-inflammatory cytokines were significantly upregulated in the kidneys. Pearson correlation analysis revealed that higher kidney ACE expression and ACE/ACE-2 ratio were positively associated with TG, TC and LDL-C levels. These data suggest that combined exposure to DEHP exposure plus a HF diet feeding may induce renal damage through overactivation of RAS.

Identification of 18β-glycyrrhetinic acid as an AGT inhibitor against LPS-induced myocardial dysfunction via high throughput screening

Sepsis induced myocardial dysfunction (SIMD) is a serious complication of sepsis. There is increasing evidence that the renin-angiotensin system (RAS) is activated in SIMD. Angiotensinogen (AGT) is a precursor of the RAS, and the inhibition of AGT may have significant cardiovascular benefits. But until now, there have been no reports of small molecule drugs targeting AGT. In this study, we designed a promoter-luciferase based system to screen for novel AGT inhibitors to alleviate SIMD. As a result of high-throughput screening, a total of 5 compounds from 351 medicinal herb‐derived natural compounds were found inhibiting AGT. 18β-glycyrrhetinic acid (18βGA) was further identified as a potent suppressor of AGT. In vitro experiments, 18βGA could inhibit the secretion of AGT by HepG2 cells and alleviate the elevated level of mitochondrial oxidative stress in cardiomyocytes co-cultured with HepG2 supernatants. In vivo, 18βGA prolonged the survival rate of SIMD mice, enhanced cardiac function, and inhibited the damage of mitochondrial function and inflammation. In addition, the results showed that 18βGA may reduce AGT transcription by downregulating hepatocyte nuclear factor 4 (HNF4) and that further alleviated SIMD. In conclusion, we provided a more efficient screening strategy for AGT inhibitors and expanded the novel role of 18βGA as a promising lead compound in rescuing cardiovascular disease associated with RAS overactivation.

Delayed Graft Function and the Renin-angiotensin System.

Delayed graft function (DGF) is a form of acute kidney injury (AKI) and a common complication following kidney transplantation. It adversely influences patient outcomes increases the financial burden of transplantation, and currently, no specific treatments are available. In developing this form of AKI, activation of the renin-angiotensin system (RAS) has been proposed to play an important role. In this review, we discuss the role of RAS activation and its contribution to the pathophysiology of DGF following the different stages of the transplantation process, from procurement and ischemia to transplantation into the recipient and including data from experimental animal models. Deceased kidney donors, whether during cardiac or brain death, may experience activation of the RAS. That may be continued or further potentiated during procurement and organ preservation. Additional evidence suggests that during implantation of the kidney graft and reperfusion in the recipient, the RAS is activated and may likely remain activated, extrapolating from other forms of AKI where RAS overactivity is well documented. Of particular interest in this setting is the status of angiotensin-converting enzyme 2, a key RAS enzyme essential for the metabolism of angiotensin II and abundantly present in the apical border of the proximal tubules, which is the site of predominant injury in AKI and DGF. Interventions aimed at safely downregulating the RAS using suitable shorter forms of angiotensin-converting enzyme 2 could be a way to offer protection against DGF.

Angiotensin II type 1 receptor-associated protein deletion combined with angiotensin II stimulation accelerates the development of diabetic kidney disease in mice on a C57BL/6 strain.

The progress in the research field of diabetic kidney disease (DKD) has been disturbed by the lack of reliable animal models. Angiotensin II (Ang II) type 1 receptor (AT1R)-associated protein (ATRAP) promotes internalization of AT1R and selectively inhibits pathological AT1R signaling. In this study, we investigated whether overactivation of the renin-angiotensin system (RAS) through a combination of ATRAP deletion with Ang II stimulation developed a progressive DKD model in C57BL/6 mice, which are resistant to the development of kidney injury. Eight-week-old male systemic ATRAP-knockout mice on the C57BL/6 strain (KO) and their littermate wild-type mice (Ctrl) were divided into five groups: 1) Ctrl, 2) Ctrl-streptozotocin (STZ), 3) KO-STZ, 4) Ctrl-STZ-Ang II, and 5) KO-STZ-Ang II. Ang II was administered for 6 weeks from 4 weeks after STZ administration. At 10 weeks after STZ administration, mice were euthanized to evaluate kidney injuries. Neither ATRAP deletion alone nor Ang II stimulation alone developed a progressive DKD model in STZ-induced diabetic C57BL/6 mice. However, a combination of ATRAP deletion with Ang II stimulation accelerated the development of DKD as manifested by overt albuminuria, glomerular hypertrophy, podocyte loss, mesangial expansion, kidney interstitial fibrosis and functional insufficiency, concomitant with increased angiotensinogen and AT1R expression in the kidneys. In STZ-induced diabetic C57BL/6 mice that are resistant to the development of kidney injury, the combination of ATRAP deletion and Ang II stimulation accelerates the development of DKD, which may be associated with intrarenal RAS overactivation.

Abstract P428: Angiotensin II Type 1 Receptor-associated Protein Deletion Combined With Angiotensin II Stimulation Exacerbates Diabetic Kidney Disease In C57BL/6 Mice

The progress in the research field of diabetic kidney disease (DKD) has been disturbed by the lack of reliable animal models. Angiotensin II (Ang II) type 1 receptor (AT1R)-associated protein (ATRAP) promotes internalization of AT1R and selectively inhibits pathological AT1R signaling. In this study, we investigated whether overactivation of the renin-angiotensin system (RAS) through a combination of ATRAP deletion with Ang II stimulation developed a progressive DKD model in C57BL/6 mice, which are resistant to the development of kidney injury. Eight-week-old male systemic ATRAP-knockout mice on the C57BL/6 strain (KO) and their littermate wild-type mice (Ctrl) were divided into five groups: 1) Ctrl, 2) Ctrl-streptozotocin (STZ), 3) KO-STZ, 4) Ctrl-STZ-Ang II, and 5) KO-STZ-Ang II. Ang II was administered for 6 weeks from 4 weeks after STZ administration. At 10 weeks after STZ administration, mice were euthanized to evaluate kidney injuries after measurements of systolic blood pressure (SBP) and urinary albumin excretion (UAE). Neither ATRAP deletion alone nor Ang II stimulation alone developed a progressive DKD model in STZ-induced diabetic C57BL/6 mice (UAE, Ctrl vs Ctrl-STZ vs KO-STZ vs Ctrl-STZ-Ang II: 10.6±2.0 vs 89.0±11.9 vs 69.1±15.0 vs 151.3±55.4 μg/day). However, a combination of ATRAP deletion with Ang II stimulation accelerated the development of DKD as manifested by overt albuminuria (UAE, KO-STZ-Ang II: 852.0±195.6 μg/day, p <0.001 vs Ctrl-STZ-Ang II), glomerular hypertrophy, podocyte loss, mesangial expansion, kidney interstitial fibrosis, and functional insufficiency, concomitant with increased angiotensinogen and AT1R expression in the kidneys. SBP levels were comparable between the Ctrl-STZ-Ang II and KO-STZ-Ang II groups (SBP, Ctrl-STZ-Ang II vs KO-STZ-Ang II: 144±4 vs 154±5 mmHg, p =0.363). In STZ-induced diabetic C57BL/6 mice, the combination of ATRAP deletion and Ang II stimulation accelerates the development of DKD by converting the C57BL/6 strain from being nephropathy-resistant to -susceptible, which may be associated with intrarenal RAS overactivation.

Diabetes-induced stimulation of the renin-angiotensin system in the rat brain cortex

Cerebrovascular disease is a threat to people with diabetes and hypertension. Diabetes can damage the brain by stimulating the renin-angiotensin system (RAS), leading to neurological deficits and brain strokes. Diabetes-induced components of the RAS, including angiotensin-converting enzyme (ACE), angiotensin-II (Ang-II), and angiotensin type 1 receptor (AT1R), have been linked to various neurological disorders in the brain. In this study, we investigated how diabetes and high blood pressure affected the regulation of these major RAS components in the frontal cortex of the rat brain. We dissected, homogenized, and processed the brain cortex tissues of control, streptozotocin-induced diabetic, spontaneously hypertensive (SHR), and streptozotocin-induced SHR rats for biochemical and Western blot analyses. We found that systolic blood pressure was elevated in SHR rats, but there was no significant difference between SHR and diabetic-SHR rats. In contrast to SHR rats, the heartbeat of diabetic SHR rats was low. Western blot analysis showed that the frontal cortexes of the brain expressed angiotensinogen, AT1R, and MAS receptor. There were no significant differences in angiotensinogen levels across the rat groups. However, the AT1R level was increased in diabetic and hypertensive rats compared to controls, whereas the MAS receptor was downregulated (p < 0.05). These findings suggest that RAS overactivation caused by diabetes may have negative consequences for the brain's cortex, leading to neurodegeneration and cognitive impairment.

The intrarenal renin-angiotensin system in hypertension: insights from mathematical modelling.

The renin-angiotensin system (RAS) plays a pivotal role in the maintenance of volume homeostasis and blood pressure. In addition to the well-studied systemic RAS, local RAS have been documented in various tissues, including the kidney. Given the role of the intrarenal RAS in the pathogenesis of hypertension, a role established via various pharmacologic and genetic studies, substantial efforts have been made to unravel the processes that govern intrarenal RAS activity. In particular, several mechanisms have been proposed to explain the rise in intrarenal angiotensin II (Ang II) that accompanies Ang II infusion, including increased angiotensin type 1 receptor (AT1R)-mediated uptake of Ang II and enhanced intrarenal Ang II production. However, experimentally isolating their contribution to the intrarenal accumulation of Ang II in Ang II-induced hypertension is challenging, given that they are fundamentally connected. Computational modelling is advantageous because the feedback underlying each mechanism can be removed and the effect on intrarenal Ang II can be studied. In this work, the mechanisms governing the intrarenal accumulation of Ang II during Ang II infusion experiments are delineated and the role of the intrarenal RAS in Ang II-induced hypertension is studied. To accomplish this, a compartmental ODE model of the systemic and intrarenal RAS is developed and Ang II infusion experiments are simulated. Simulations indicate that AT1R-mediated uptake of Ang II is the primary mechanism by which Ang II accumulates in the kidney during Ang II infusion. Enhanced local Ang II production is unnecessary. The results demonstrate the role of the intrarenal RAS in the pathogenesis of Ang II-induced hypertension and consequently, clinical hypertension associated with an overactive RAS.

An overview of the effects of combination therapy with a renin-angiotensin system blocker plus a calcium channel blocker on glucose homeostasis in non-diabetic hypertensive patients

Arterial hypertension (AH) is a major cardiovascular risk factor and often coexists with insulin resistance. Insulin resistance impairs glucose homeostasis and has been associated with the development of new-onset type 2 diabetes mellitus (T2D). Overactivity of renin-angiotensin system (RAS) mediated by angiotensin II adversely affects glucose homeostasis. The blockade of RAS with the use of angiotensin converting enzyme inhibitors (ACEi) or angiotensin receptor blockers (ARBs) has been associated with beneficial effects on glucose metabolism. On the other hand, calcium channel blockers (CCBs) have been reported to exert a metabolic neutral effect. By contrast, the use of diuretics and beta-blockers has been shown to have an overall negative effect on glucose metabolism. Current ESH/ESC treatment guidelines recommend the use of fixed single-pill combinations of RAS blockers with either CCBs or thiazide/thiazide-like diuretics in hypertensive patients with grade 1 AH and high cardiovascular risk, or greater. Nonetheless, considering the patient’s medical history and comorbidities, antihypertensive treatment should be carefully tailored. To this, hypertensive patients at risk of developing new-onset T2D, with impaired fasting glucose (IFG) or impaired glucose tolerance (IGT), should be preferentially treated with either monotherapy or combined treatment using agents which do not affect or worsen glucose homeostasis.

Arachidonyl‐2‐ Chlorethylamide alters the Gene Expression of Renin Angiotensin System Components in Primary Cultures of Astrocytes

ObjectiveTo determine the of effect Arachidonyl‐2′‐chloroethylamide (ACEA) on the gene expression of renin angiotensin system (RAS) components in rat astrocytes.BackgroundWe have previously shown that activation of the Angiotensin type 1 receptor (AT1R) altered the expression of the cannabinoid receptor type 1 (CB1R) in rat astrocytes. Activation of the CB1R can potentially have synergistic or antagonistic effects on AT1R actions. Studies have shown that an overactive RAS contributes to pathological conditions. Thus, we determined the molecular effects of activation of the CB1R by ACEA on RAS components. To investigate this, we determined ACEA and Angiotensin (Ang) II effects alone, then combined, on the mRNA levels of RAS components in astrocytes isolated from the cerebellum of Wistar and Spontaneously hypertensive rats (SHRs). CB1R and RAS components are expressed in the cerebellum.MethodsAstrocytes were prepared from 2‐3 days rat pups. Quiescent cells were treated with 10nM ACEA and or 100nM Ang II for 12‐24 hours. mRNA levels of RAS components were measured by quantitative polymerase chain reaction.ResultsAng II predominantly downregulated RAS components in both Wistar and SHR astrocytes at the time points examined. ACEA downregulated AT1R mRNA in Wistar, increased angiotensinogen mRNA in both Wistar and SHR astrocytes, and increased angiotensin converting enzyme mRNA in SHR astrocytes. The effects of combined treatments were similar to Ang II single treatments.ConclusionCB1R activation altered the mRNA levels of RAS components but did not appear to have considerable impact on Ang II’s effects on these components. Since mRNA levels might not correlate with protein levels, determination of protein levels and/or activity would complement these findings.

Intrarenal Renin‐Angiotensin System and its Role in Angiotensin II‐Induced Hypertension: Insights from Computational Modelling

The kidney expresses all and independently regulates most elements of the renin‐angiotensin system (RAS). Importantly, the inappropriate activation of intrarenal RAS, as indicated by the local accumulation of angiotensin II (Ang II), has been linked to Ang II‐induced hypertension. Various Ang II and angiotensin type 1 receptor (AT1R)‐dependent positive feedback systems have been documented in the literature. However, their functional importance to the overactivation of the intrarenal RAS have yet to be lineated. The goal of this study is to identify the key mechanisms actuating the intrarenal accumulation of Ang II during hypertension induced by Ang II infusion. We hypothesize that the upregulation of tubular epithelial AT1Rs is the primary mediator of intrarenal RAS overactivation following Ang II infusion. To test this hypothesis, computational models of the intrarenal RAS and the AT1R‐blocker Losartan were created, coupled, and used to simulate Ang II‐infusion experiments in the absence and presence of this anti‐hypertensive therapy. Model simulations indicate that total renal Ang II concentrations rise once Ang II starts accumulating within tubular epithelial cells and that the upregulation of local AT1R expression, not Ang II production, is required to produce this effect. Indeed, the removal of this positive feedback and the administration of Losartan both prevented the intrarenal accumulation of Ang II. Since the simulated onset of Ang II accumulation in tubular epithelial cells coincides exactly with the experimental inception of Ang II‐dependent hypertension, we conclude that the stimulation of sodium reabsorption mediated by local Ang II is likely key to the development of Ang II‐induced hypertension.

Chronic Renin-Angiotensin System Activation Induced Neuroinflammation: Common Mechanisms Underlying Hypertension and Dementia?

Hypertension is a major risk factor for the pathogenesis of vascular dementia and Alzheimer's disease. Chronic activation of the renin-angiotensin system (RAS) contributes substantially to neuroinflammation. We propose that neuroinflammation arising from chronic RAS activation can initiate and potentiate the onset of hypertension and related dementia. Neuroinflammation induced by chronic activation of the RAS plays a key role in the pathogenesis of dementia. Increased levels of pro-inflammatory cytokines tumor necrosis factor (TNF)-α, interleukin (IL)-1β, and transforming growth factor (TGF)-β have been reported in brain tissue of vascular dementia patients and animal models of vascular dementia induced by either angiotensin II infusion or transverse aortic coarctation. It is proposed that neuronal cell death and synaptic dysfunction induced by neuroinflammation lead to cognitive impairment in dementia. The neuroprotective RAS pathway, regulated by angiotensin-converting enzyme 2 (ACE2) which converts angiotensin II into angiotensin-(1-7), can attenuate hypertension and dementia. Furthermore, the use of anti-hypertensive medications in preventing dementia or cognitive decline in hypertensive patients and animal models of dementia have mostly been beneficial. Current evidence suggests a strong link between RAS induced neuroinflammation and the onset of hypertension and dementia, which warrants further investigation. Strategies to counteract an overactive RAS and enhance the neuroprotective arm of the RAS may help prevent or improve cognitive impairment associated with hypertension.

Age and genotype dependent erythropoietin protection in COVID-19.

Erythropoietin (EPO) is the main mediator of erythropoiesis and an important tissue protective hormone that appears to mediate an ancestral neuroprotective innate immune response mechanism at an early age. When the young brain is threatened-prematurity, neonatal hyperbilirubinemia, malaria- EPO is hyper-secreted disproportionately to any concurrent anemic stimuli. Under eons of severe malarial selection pressure, neuroprotective EPO augmenting genetic determinants such as the various hemoglobinopathies, and the angiotensin converting enzyme (ACE) I/D polymorphism, have been positively selected. When malarial and other cerebral threats abate and the young child survives to adulthood, EPO subsides. Sustained high ACE and angiotensin II (Ang II) levels through the ACE D allele in adulthood may then become detrimental as witnessed by epidemiological studies. The ubiquitous renin angiotensin system (RAS) influences the α-klotho/fibroblast growth factor 23 (FGF23) circuitry, and both are interconnected with EPO. Here we propose that at a young age, EPO augmenting genetic determinants through ACE D allele elevated Ang II levels in some or HbE/beta thalassemia in others would increase EPO levels and shield against coronavirus disease 2019, akin to protection from malaria and dengue fever. Human evolution may use ACE2 as a “bait” for severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) to gain cellular entry in order to trigger an ACE/ACE2 imbalance and stimulate EPO hypersecretion using tissue RAS, uncoupled from hemoglobin levels. In subjects without EPO augmenting genetic determinants at any age, ACE2 binding and internalization upon SARS-CoV-2 entry would trigger an ACE/ACE2 imbalance, and Ang II oversecretion leading to protective EPO stimulation. In children, low nasal ACE2 Levels would beneficially augment this imbalance, especially for those without protective genetic determinants. On the other hand, in predisposed adults with the ACE D allele, ACE/ACE2 imbalance, may lead to uncontrolled RAS overactivity and an Ang II induced proinflammatory state and immune dysregulation, with interleukin 6 (IL-6), plasminogen activator inhibitor, and FGF23 elevations. IL-6 induced EPO suppression, aggravated through co-morbidities such as hypertension, diabetes, obesity, and RAS pharmacological interventions may potentially lead to acute respiratory distress syndrome, cytokine storm and/or autoimmunity. HbE/beta thalassemia carriers would enjoy protection at any age as their EPO stimulation is uncoupled from the RAS system. The timely use of rhEPO, EPO analogs, acetylsalicylic acid, bioactive lipids, or FGF23 antagonists in genetically predisposed individuals may counteract those detrimental effects.

Vitamin D association with the renin angiotensin system in polycystic ovary syndrome

Vitamin D deficiency is a negative endocrine renin-angiotensin system (RAS) modulator and PCOS women are often vitamin D deficient, leading to RAS overactivation in PCOS.A cross-sectional study was performed in 99 PCOS and 68 control women who presented sequentially. Circulating plasma levels of RAS proteins (Angiotensin-converting enzyme 2 (ACE2), renin and angiotensinogen) were measured by Slow Off-rate Modified Aptamer (SOMA)-scan and 25-hydroxyvitamin D [25(OH)D] was measured by tandem mass spectroscopy.The RAS system was found to be overactivated in the PCOS women compared to non-PCOS control women with increased renin and decreased angiotensinogen (p < 0.05); 25-hydroxyvitamin D was also significantly lower in the PCOS group (p < 0.0001). In PCOS women, plasma renin was increased in vitamin D deficient and insufficient groups compared with the vitamin D sufficient group (p < 0.005), but did not differ across non-PCOS control subgroups. In non-PCOS controls, plasma ACE2 decreased from vitamin D insufficiency to deficiency (p < 0.05). Angiotensinogen was not different across the vitamin D sufficiency, insufficiency and deficiency strata for either PCOS or non-PCOS controls.These data show that RAS activation through increased plasma renin levels was seen in vitamin D insufficient and deficient PCOS subjects compared to non-PCOS control women. In addition, decreased plasma ACE2 levels were seen in vitamin D deficiency in non-PCOS controls, which may predispose these vitamin D deficient subjects to increased cardiovascular risk and susceptibility to infectious agents such as COVID-19 where this is a risk factor.

Abstract MP09: Effect Of Western Diet On Renal Transcriptome Of Hypertensive Mice Overexpressing Human Angiotensin Receptor Type 1

Over-expression of human angiotensin-II receptor type1 (hAT1R) may cause pathological outcomes due to overactivation of renin-angiotensin system. Transgenic (TG) mice containing Hap-I (hypertensive genotype) of human hAT1R gene are more prone to develop metabolic syndrome disorders as compared to TG mice with Hap-II (normotensive genotype). This gene variant associated risk of hypertension together with Western diet and aging may lead to renal disorders. However, mechanisms underlying this process are not well examined. For this purpose, we studied the renal gene expression alterations in aged TG mice containing either Hap-I or Hap-II of hAT1R gene. Aged mice (20-24 months of age) were maintained on a regular diet or high fat diet with 2% NaCl (Western diet, WD) for 16 weeks. On a regular diet, aged Hap-I mice presented higher (~9 mmHg) systolic blood pressure with respect to age-matched Hap-II animals. Following administration of Western diet, blood pressure increased in both groups of mice, but to a larger extent in Hap-I animals (~15 mmHg in comparison to ~7 mmHg in Hap-II). Aged Hap-I mice on Western diet showed increased renal fibrosis. RNA-seq data from renal tissue of Hap-I aged mice revealed that WD significantly altered the expression of &gt;400 genes (p-adj. &lt;0.05). Bioinformatics analysis (Qiagen IPA software) identified major alterations in main canonical pathways involved in renal function and oxidative damage. These changes in turn resulted in kidney failure, renal tubular injury, and renal proliferation. In addition, post WD treatment, RNA seq. analysis from Hap-I and Hap-II kidneys also reveals haplotype specific regulation of genes associated with blood pressure regulation and kidney disorders. Overall, these results indicate that Western diet promotes hypertension and fibrosis in the kidneys of aged mice. These alterations are paralleled by perturbation of renal transcriptional profile. Overall, these studies will assist in the identification of novel mechanisms and molecules involved in hypertension and associated kidney pathophysiology.