AT1 Receptor Blocker Research Articles

Valsartan Attenuated Homocysteine-Induced Impaired Autophagy and ER Stress in Human Umbilical Vein Endothelial Cells.

Hyperhomocysteinemia is a risk factor for various cardiovascular diseases. However, the mechanism underlying homocysteine- (Hcy-) induced vascular injury remains unclear. The purpose of the present study was to examine a potential mechanism by which Hcy induced injury in human umbilical vascular endothelial cells (HUVEC). The protein abundance of autophagy-related markers was markedly decreased after Hcy treatment, which was associated with endoplasmic reticulum (ER) stress and apoptosis in HUVECs. Protein expression level of angiotensin II type 1 receptor (AT1 receptor) was dramatically increased in response to Hcy. Valsartan, an AT1 receptor blocker, improved autophagy and prevented ER stress and apoptosis in HUVECs treated with Hcy. Consistent with this, silence of AT1 receptor with siRNA decreased the protein abundance of ER stress markers, prevented apoptosis, and promoted autophagy in HUVECs. Inhibition or knockdown of AT1 receptor was shown to be associated with suppression of p-GSK3β/GSK3β-p-mTOR/mTOR signaling pathway. Additionally, inhibition of autophagy by 3-MA aggravated Hcy-induced apoptosis, while amelioration of ER stress by 4-PBA prevented Hcy-induced injury in HUVECs. Hcy-induced HUVEC injury was likely attributed to AT1 receptor activation, leading to impaired autophagy, ER stress, and apoptosis.

Unlocking the protective potential of the angiotensin type 2 receptor (AT2R) in acute lung injury and age-related pulmonary dysfunction

Despite its known importance in the cardiovascular system, the specific role and impact of the angiotensin type 2 receptor (AT2R) in lung physiology and pathophysiology remain largely elusive. In this study, we highlight the distinct and specialized lung-specific roles of AT2R, primarily localized to an alveolar fibroblast subpopulation, in contrast to the angiotensin type 1 receptor (AT1R), which is almost exclusively expressed in lung pericytes. Evidence from our research demonstrates that the disruption of AT2R (AT2R-/y), is associated with a surge in oxidative stress and impaired lung permeability, which were further intensified by Hyperoxic Acute Lung Injury (HALI). With aging, AT2R-/y mice show an increase in oxidative stress, premature enlargement of airspaces, as well as increased mortality when exposed to hyperoxia as compared to age-matched WT mice. Our investigation into Losartan, an AT1R blocker, suggests that its primary HALI lung-protective effects are channeled through AT2R, as its protective benefits are absent in AT2R-/y mice. Importantly, a non-peptide AT2R agonist, Compound 21 (C21), successfully reverses lung oxidative stress and TGFβ activation in wild-type (WT) mice exposed to HALI. These findings suggest a possible paradigm shift in the therapeutic approach for lung injury and age-associated pulmonary dysfunction, from targeting AT1R with angiotensin receptor blockers (ARBs) towards boosting the protective function of AT2R.

Antihypertensive treatment of end-stage renal disease patients on hemodialysis does not alter circulating ACE and ACE2 activity and angiotensin peptides

Cardiovascular diseases (CVD) are the main causes of death in hemodialysis patients, representing a public health challenge. We investigated the effect of different antihypertensive treatments on circulating levels of renin-angiotensin system (RAS) components in end-stage renal disease (ESRD) patients on hemodialysis. ESRD patients were grouped following the prescribed antihypertensive drugs: ß-blocker, ß-blocker+ACEi and ß-blocker+AT1R blocker. ESDR patients under no antihypertensive drug treatment were used as controls. Blood samples were collected before hemodialysis sessions. Enzymatic activities of the angiotensin-converting enzymes ACE and ACE2 were measured through fluorescence assays and plasma concentrations of the peptides Angiotensin II (Ang II) and Angiotensin-(1–7) [Ang-(1–7)] were quantified using mass spectrometry (LC-MS/MS). ACE activity was decreased only in the ß-blocker+ACEi group compared to the ß-blocker+AT1R, while ACE2 activity did not change according to the antihypertensive treatment. Both Ang II and Ang-(1–7) levels also did not change according to the antihypertensive treatment. We concluded that the treatment of ESRD patients on hemodialysis with different antihypertensive drugs do not alter the circulating levels of RAS components.

Abstract 12379: AT1-AA (Angiotensin II Type 1 Receptor Autoantibody) Aggravates Abdominal Aortic Aneurysm Through Activating AT1R/Gq Allosterically

Background: Angiotensin II type 1 receptor (AT1R) is a key mediator of the rennin-angiotensin-aldosterone system (RAAS). Autoantibodies against AT1R (AT1-AA), as an activator of AT1R, was found in abdominal aortic aneurysm (AAA) related diseases, such as hypertension and atherosclerosis. However, the role of AT1-AA in AAA is still unclear. Methods: We used angiotensin II (AngII)-infused male ApoE -/- mice model of AAA. Ultrasonography measurement, histological assessment, western blot, qRT-PCR and RNA sequencing were used to assess the severity of AAA. Nanoluciferase-based bioluminescence resonance energy transfer (NanoBRET) assay, radio assay of receptors and homogeneous time resolved fluorescence (HTRF) were used to measure the activation feature of AT1R and the affinity of AT1R with AT1-AA or AngII. Results: AT1-AA increased the incidence of AAA (vehicle vs. AT1-AA: 47% vs. 81%; P<0.05), which can be attenuated by AT1R blocker Candesartan. Furthermore, AT1-AA greatly increased severity of AAA, due to augmenting the diameter of abdominal aorta and the degradation of elastin by enhancing the expression of MMP2/9 (matrix metalloproteinase) and the accumulation of inflammatory cells. And AT1-AA could induce phenotypic switch of smooth muscle cells. But AT1-AA does not increase the plasma level of AngII. Mechanically, AT1-AA enhanced AngII-induced recruitment of Gq, but not β-arrestin1/2. AT1-AA potentiated the AngII-mediated Gq-dependent inositol phosphate (EC50 shift=3.829 nM) and the activation of PKC and ERK1/2. The responses were abolished by AT1R blocker and Gq inhibitor. The affinity of AT1-AA for the AT1R in cells was calculated to be 75.49 ± 4.7 nM. AT1-AA cannot compete for the binding of AngII to AT1R. However, AT1-AA significantly potentiated the binding of AngII with AT1R allosterically (mean K D ± SEM: 34.11 ± 0.9765 nM at 0 nM AT1-AA vs. 25.63 ± 1.826 nM at 100 nM AT1-AA; p<0.05), without altering the maximum binding. Conclusion: AT1-AA, as an allosteric activator of AT1R, promotes AAA through increasing the affinity of AngII with AT1R and activating Gq biasedly.

Angiotensin II reduces glyoxalase 1 activity and expression in vascular smooth muscle cells: Implications for diabetic vascular complications.

Angiotensin II (Ang II), a key mediator of vascular diseases, is linked to methylglyoxal (MGO) formation, a by-product of glucose metabolism implicated in vascular complications. The glyoxalase system, consisting of glyoxalase 1 (Glo1) and reduced glutathione (GSH), is responsible for detoxifying MGO. This study investigated the effect of Ang II on Glo1 activity and expression in vascular smooth muscle cells (VSMCs). Primary VSMCs were isolated from rat aortas and exposed to Ang II under standard or high glucose conditions. We examined Glo1 activity, expression, intracellular GSH, and methylglyoxal-derived hydroimidazolone 1 (MG-H1) levels. We also analyzed the expressions of nuclear factor-κB (NF-κB) p65 and nuclear factor erythroid 2-related factor 2 (Nrf2) as potential regulators of Glo1 expression. The results demonstrated that Ang II reduced Glo1 activity, expression, and GSH levels while increasing MG-H1 levels in VSMCs. Telmisartan and irbesartan, AT1R blockers, restored Glo1 activity, expression, and GSH levels and alleviated MG-H1 levels. Treatment with AT1R blockers or inhibitors targeting signaling pathways involved in Ang II-induced responses mitigated these effects. High glucose exacerbated the reduction in Glo1 activity and expression. In conclusion, this study provides evidence that Ang II reduces Glo1 activity and expression in VSMCs, which may contribute to developing vascular complications in diabetes. AT1R blockers and inhibitors targeting specific signaling pathways show potential in restoring Glo1 function and mitigating MGO-associated damage. These findings highlight the complex interactions between RAS, MGO, and vascular diseases, highlighting potential therapeutic targets for diabetic vascular complications.

Ameliorating Effect of Azilsartan on Cisplatin -Induced Ocular Toxicity in Male Rats

Objective: To evaluate the protective effect of Azilsartan against Cisplatin-induced ocular damage by ameliorating the oxidative stress and inflammation status, as well as to compare two different doses of Azilsartan to the Resveratrol. Fifty-six male Wister albino-rats, weighing 270±30 g. The rats were allocated at random into seven groups (n=8 per group), as follows: Group 1 (Healthy control) was given 0.5% CMC orally for the 14 days. Group 2 (Positive control) was given a single injection of 7mg/kg Cisplatin intraperitoneally and then 0.5% CMC orally for the following 14 days. Group 3 was given 3.5mg/kg Azilsartan orally for the following 14 days. Group 4 was given 7mg/kg Azilsartan orally for the following 14 days. Group 5 was given a single injection of 7mg/kg Cisplatin intraperitoneally and then 3.5mg/kg Azilsartan orally for the following 14 days. Group 6 was given a single injection of 7mg/kg Cisplatin intraperitoneally and then 7mg/kg Azilsartan orally for the following 14 days. Group 7 (standard) was given 25mg/kg/day Resveratrol orally for following 14 days.Azilsartan at low dose (3.5mg/kg) showed a significant reduction in IL-1β pro-inflammatory marker level, whereas there was no significant effect on MDA and SOD levels in cisplatin-induced ocular damage. Histologically, there were significant reduction in inflammatory exudates, edema, and inflammatory cells infiltration with both doses. Additionally, there were no significant difference among Azilsartan only received groups and Resveratrol group. Azilsartan shows a promising anti-inflammatory effect in ocular tissue in Cisplatin experimental rat models by significantly reducing IL-1β overexpression, through its potent AT1receptor blocking effect. However, Azilsartan showed a slight effect on MDA levels and marginal effect on SOD levels in Cisplatin-induced ocular toxicity. Furthermore, Azilsartan at a low dose slightly mimicked the effect of resveratrol on normal ocular tissue, suggesting an advancement in prophylactic application in ocular injury.
 Keywords: Azilsartan, IL-1β, Cisplatin, Ocular Toxicity

Drug therapy targeting angiotensin II type 1 receptors in brain against frequent urination

The production of angiotensin II (Ang II) in the brain plays important roles as neurotransmitter and neuropeptide. Central Ang II is involved in regulating various physiological processes, such as blood pressure and water homeostasis, via Ang II type 1 (AT1) receptors. We have demonstrated that Ang II induces frequent urination via AT1 receptors in the brain even at doses that does not seem to affect the blood pressure in animal experiment. Intracerebroventricular administration of Ang II was also found to reduce the bladder capacity without affecting the maximum voiding pressure, post voiding residual urine volume or voiding efficiency. Additionally, the activation of AT1 receptor downstream signal pathway (phospholipase C/protein kinase C/NADPH oxidase/superoxide anion) and suppression of GABAergic nervous system in the brain are involved in the mechanism underlying the central Ang II-inducted frequent urination. AT1 receptor blockers (ARBs) have been widely used to treat hypertension. We demonstrated that peripherally administered ARBs telmisartan, which can penetrate blood-brain barrier, exerted an inhibitory effect on central Ang II-inducted frequent urination. We present the possible drug therapy targeting AT1 receptors in the brain against frequent urination on the results obtained from our recent research work.

Astrocytic AT1R deficiency ameliorates Aβ-induced cognitive deficits and synaptotoxicity through β-arrestin2 signaling.

Alzheimer's disease (AD) seriously influences human health, and there is no effective treatment to prevent or cure AD. Recent studies have shown that angiotensin II type 1 receptor (AT1R) blockers significantly reduce the prevalence of AD, while the precise role and mechanism of AT1R in AD remain obscure. In this study, for the first time, we identified that astrocytic but not neuronal AT1R levels were significantly increased in AD model rats and found that astrocyte-specific knockout of AT1R significantly ameliorated amyloid β (Aβ)-induced cognitive deficits and synaptotoxicity. Pretreating astrocytes with an AT1R blocker also alleviated Aβ-induced synaptotoxicity in the coculture system of hippocampal neurons and astrocytes. Moreover, AT1R could directly bind to Aβ1-42 and activate the astrocytic β-arrestin2 pathway in a biased manner, and biased inhibition of the astrocytic AT1R/β-arrestin2 pathway relieved Aβ-induced neurotoxicity. Furthermore, we demonstrated that astrocytic AT1R/β-arrestin2 pathway-mediated synaptotoxicity was associated with the aggregation of autophagosomes, which triggered the disordered degradation of Aβ. Our findings reveal a novel molecular mechanism of astrocytic AT1R in Aβ-induced neurodegeneration and might contribute to establishing new targets for AD prevention and therapy.

Abstract 100: Activation Of The Skin Renin-angiotensin System Contributes To The Development Of Hypertension

Recent studies suggest skin vasoconstriction and sodium (Na + ) accumulation are associated with hypertension. The skin has a localized renin-angiotensin system (RAS), a key regulator of blood pressure (BP), but its functional role remains unclear. This study aims to investigate the role of skin RAS in hypertension. Skin tissue from 32 individuals, including patients with hypertension, was used to evaluate the association between skin RAS component expression and office BP. Among the RAS components, skin expression of ATRAP (type-1 angiotensin II (Ang II) receptor (AT1R)-associated protein), which selectively inhibits pathological AT1R signaling, was inversely correlated with systolic and diastolic BP (R 2 = 0.12, P < 0.05; R 2 = 0.20, P < 0.05, respectively). To investigate the causal relationship between skin ATRAP and hypertension, mice lacking ATRAP in skin keratinocytes (KO: K14 Cre ;ATRAP flox ) were generated. Telemetry analysis showed that Ang II (500 ng/kg/min)-induced hypertension and related cardiac hypertrophy was exacerbated in KO mice compared to control mice (mean BP, 123.3 ± 4.7 vs. 132.6 ± 3.1 mmHg, n = 5, P < 0.05; heart/body weight, 4.7 ± 0.4 vs. 5.2 ± 0.7, n = 11-12, P < 0.05). KO mice showed increased skin expression of angiotensinogen and AT1R ( P < 0.05 for each; n = 11-12), while those in the kidneys and heart were similar in both groups, suggesting skin-specific enhancement of tissue RAS activity. AT1R blocker treatment eliminated exaggerated Ang II-induced hypertension and enhancement of skin RAS activity in KO mice. Sympathetic nerve activity, plasma volume, and skin Na + content did not differ between the groups. In KO mice, urine volume per water intake increased ( P < 0.001), despite similar body weight changes in both groups, suggesting decreased extra-renal water loss. Consistent with these findings, skin blood flow (22.6 ± 3.8 vs. 16.5 ± 4.5 AU, P < 0.001) and transepidermal water loss (9.0 ± 1.9 vs. 6.5 ± 1.9 g/m 2 /h, P < 0.01) were decreased in KO mice. These differences, including exaggerated Ang II-induced hypertension, were eliminated by skin vasodilation via body temperature elevation. In conclusion, enhanced skin RAS activity contributes to BP elevation via skin vasoconstriction. Skin RAS may be a novel target for treating hypertension.

Novel AT2R agonist, β-Pro7Ang III, is cardio- and vaso-protective in diabetic spontaneously hypertensive rats

Stimulation of the angiotensin II type 2 receptor (AT2R) evokes protective effects in various cardiovascular diseases. Thus, this study aimed to investigate the effects of AT2R stimulation, with or without AT1R blockade, in a model of hypertension with concomitant type 1 diabetes mellitus (T1DM). Spontaneously hypertensive rats (SHRs) were given either citrate or a single dose of streptozotocin (STZ; 55 mg/kg, i.p.) to induce diabetes. After 4 weeks of diabetes, animals were administered either a vehicle (saline), AT2R agonist, β-Pro7Ang III (0.1 mg/kg/day via osmotic mini-pump), AT1R blocker, candesartan (2 mg/kg/day via drinking water), or a combination of both for a further 8 weeks. β-Pro7Ang III treatment had no effect on blood pressure, but attenuated the significant increase in cardiac interstitial collagen and protein expression of fibrotic and inflammatory markers, and superoxide levels that was evident in diabetic SHRs. These effects were not observed with candesartan, despite its blood pressure lowering effects. Although β-Pro7Ang III had no effect on aortic fibrosis, it significantly attenuated MCP-1 protein expression and superoxide levels when compared to both the non-diabetic and diabetic SHRs, to a similar extent as candesartan. In both the heart and vasculature, the effects of β-Pro7Ang III in combination with candesartan were similar to those of β-Pro7Ang III alone, and superior to candesartan alone. It was concluded that in hypertension with concomitant diabetes, AT2R stimulation with a novel ligand alone, or in combination with AT1R blockade, improved the cardiac and vascular structural changes that were strongly associated with inflammation and oxidative stress, independent of blood pressure regulation.

#4590 A PROSPECTIVE STUDY OF EPLERENONE IN THE TREATMENT OF PATIENTS WITH GLOMERULONEPHRITIS

Abstract Background and Aims Clinical evidence suggests that high aldosterone plasma levels contributes to progressive kidney disease. Although administration of angiotensin-converting enzyme inhibitors (ACEi) or angiotensin type 1 receptor blockers (ARB) can delay kidney disease progression, plasma aldosterone levels may increase to a significant extent even after the initiation of such treatment. Thus, administration of mineralocorticoid receptor antagonists such as spironolactone, on top of treatment with ACEi or ARBs has been shown to reduce albuminuria and slow down kidney function deterioration. Nevertheless, there is scanty evidence on the effect of eplerenone in patients with kidney disease due to glomerulonephritis. Method In this prospective open-label study, we evaluated the effects of eplerenone in patients with biopsy proven glomerulonephritis already treated with ACEi or ARBs. Patients were divided in the active treatment arm, who received 25 mg of eplerenone daily besides ACEi or ARBs, and a control group that received only standard treatment. Protein excretion in 24 h urine, eGFR, blood pressure and serum K+ levels were measured at 3, 6 and 12 months after study initiation. Results Out of 83 patients who were screened, 62 patients were included in the final analysis. Twenty-six received eplerenone and 36 were treated with ACEi (25) or ARBs (9) alone (controls). After 1 year of treatment with eplerenone, proteinuria decreased from 1597 to 1145 mg/24 h while it did not alter in controls. Eplerenone reduced proteinuria especially in those with baseline proteinuria of >1000 mg/24 h. Kidney function remained stable in patients treated with eplerenone (from eGFR: 82.51±22.6 to 84.68±31.7 ml/min/1.73 m2, p = 0.8) and showed a non significant deterioration in controls (from eGFR: 68.39±26.2 to 66.63±29.8 ml/min/1.73 m2, p = 0.08). Systolic blood pressure was significantly reduced in the active treatment arm (from 128.1±17.5 to 126.3±10.93 mmHg, p = 0.03), whereas eplerenone did not increase serum K+ levels or had any other significant adverse effect. Conclusion Administration of eplerenone at a dose of 25 mg/day, on top of treatment with ACEi or ARBs, can further reduce proteinuria and systolic blood pressure in patients with chronic glomerulonephritis.

Inflammatory mediators act at renal pericytes to elicit contraction of vasa recta and reduce pericyte density along the kidney medullary vascular network.

Introduction: Regardless of initiating cause, renal injury promotes a potent pro-inflammatory environment in the outer medulla and a concomitant sustained decrease in medullary blood flow (MBF). This decline in MBF is believed to be one of the critical events in the pathogenesis of acute kidney injury (AKI), yet the precise cellular mechanism underlying this are still to be fully elucidated. MBF is regulated by contractile pericyte cells that reside on the descending vasa recta (DVR) capillaries, which are the primary source of blood flow to the medulla. Methods: Using the rat and murine live kidney slice models, we investigated the acute effects of key medullary inflammatory mediators TNF-α, IL-1β, IL-33, IL-18, C3a and C5a on vasa recta pericytes, the effect of AT1-R blocker Losartan on pro-inflammatory mediator activity at vasa recta pericytes, and the effect of 4-hour sustained exposure on immunolabelled NG2+ pericytes. Results and discussion: Exposure of rat and mouse kidney slices to TNF-α, IL-18, IL-33, and C5a demonstrated a real-time pericyte-mediated constriction of DVR. When pro-inflammatory mediators were applied in the presence of Losartan the inflammatory mediator-mediated constriction that had previously been observed was significantly attenuated. When live kidney slices were exposed to inflammatory mediators for 4-h, we noted a significant reduction in the number of NG2+ positive pericytes along vasa recta capillaries in both rat and murine kidney slices. Data collected in this study demonstrate that inflammatory mediators can dysregulate pericytes to constrict DVR diameter and reduce the density of pericytes along vasa recta vessels, further diminishing the regulatory capacity of the capillary network. We postulate that preliminary findings here suggest pericytes play a role in AKI.

Clarification of hypertension mechanisms provided by the research of central circulatory regulation.

Sympathoexcitation, under the regulatory control of the brain, plays a pivotal role in the etiology of hypertension. Within the brainstem, significant structures involved in the modulation of sympathetic nerve activity include the rostral ventrolateral medulla (RVLM), caudal ventrolateral medulla (CVLM), nucleus tractus solitarius (NTS), and paraventricular nucleus (paraventricular). The RVLM, in particular, is recognized as the vasomotor center. Over the past five decades, fundamental investigations on central circulatory regulation have underscored the involvement of nitric oxide (NO), oxidative stress, the renin-angiotensin system, and brain inflammation in regulating the sympathetic nervous system. Notably, numerous significant findings have come to light through chronic experiments conducted in conscious subjects employing radio-telemetry systems, gene transfer techniques, and knockout methodologies. Our research has centered on elucidating the role of NO and angiotensin II type 1 (AT1) receptor-induced oxidative stress within the RVLM and NTS in regulating the sympathetic nervous system. Additionally, we have observed that various orally administered AT1 receptor blockers effectively induce sympathoinhibition by reducing oxidative stress via blockade of the AT1 receptor in the RVLM of hypertensive rats. Recent advances have witnessed the development of several clinical interventions targeting brain mechanisms. Nonetheless, Future and further basic and clinical research are needed.

GENETIC DELETION OR PHARMACOLOGICAL INHIBITION OF THE SODIUM AND HYDROGEN EXCHANGER 3 IN THE PROXIMAL TUBULES OF THE KIDNEY ATTENUATES ANGIOTENSIN II AND SALT-INDUCED HYPERTENSION

Objective: According to American Heart Association, ∼46% of American adults have hypertension and ∼13 of these patients develop poorly controlled hypertension even treated with >3 - 4 different classes of antihypertensive drugs. The mechanisms underlying poorly controlled hypertension remain incompletely understood. The present study tested the proof of concept hypothesis that the sodium and hydrogen exchanger 3 (NHE3) in the proximal tubules of the kidney may be a therapeutic target of angiotensin II (Ang II) and salt-induced hypertension in mice. Design and method: To test the hypothesis, we generated a new mouse model with deletion of NHE3 selectively in the proximal tubules of the kidney using the iL-Sglt2-Cre and Nhe3-floxed approach (PT-Nhe3-/-). Adult male wildtype (WT) and PT-Nhe3-/- mice were infused without or with a low dose of Ang II (0.5 mg/kg/day, i.p., via minipump) and maintained on 2% high salt diet for 2 weeks to induce Ang II- and salt-dependent hypertension. Animals were then treated without or with an absorbable NHE3 inhibitor AVE-0657 for 2 weeks (20 mg/kg/day, p.o., a gift of Sanofi-Aventis). Results: Under basal conditions, deletion of NHE3 selectively in the proximal tubules of the kidney decreased basal systolic and mean blood pressure by ∼13 ± 3 mmHg in PT-Nhe3-/- mice (P < 0.01 vs. WT), whereas inhibition of NHE3 primarily in the proximal tubules with AVE-0657 lowered basal blood pressure by ∼7 ± 2 mmHg in WT mice (P < 0.05 vs. control WT). Ang II and high salt treatment significantly increased blood pressure by 26 ± 5 mmHg in WT mice (P < 0.01), which was significantly attenuated in PT-Nhe3-/- mice (P < 0.01). Ang II and salt-induced hypertension in WT mice was also significantly attenuated by NHE3 inhibition in the kidney proximal tubules with AVE-0657 (P < 0.01). Concurrent treatment with losartan, an AT1 receptor blocker (20 mg/kg/day, p.o.), completely lowered blood pressure to control level in WT mice (P < 0.05). AVE-0657 had no further effect on blood pressure in PT-Nhe3-/- mice. Conclusions: In conclusion, our results support the proof of concept hypothesis that NHE3 in the proximal tubules of the kidney may be a therapeutic target of Ang II and salt-induced hypertension.

GINSENOSIDE-RG3 ENHANCES SOME ANTIHYPERTENSIVE AGENTS-INDUCED INHIBITION OF ADRENAL CATECHOLAMINE SECRETION

Objective: The present study was designed to examine the influence of ginsenoside-Rg3 (Rg3), one of panaxadiol-type saponins, which is isolated from Korean Panax ginseng, in some antihypertensive agents-induced inhibition on the secretion of catecholamines (CA) from the perfused rat adrenal medullae. Design and method: The adrenal gland was isolated and perfused with Krebs-bicarbonate. CA was measured directly by using the fluorospectrophotometer. Results: Ginsenoside-Rg3 (3∼30 μM) during perfusion into an adrenal vein for 90 min significantly inhibited the CA secretion evoked by acetylcholine (ACh). Also, olmesartan (AT1 receptor blocker, 15 μM), simvastatin (an inhibitor of 3-hydroxy-3-methylglutaryl coenzyme A reductase, 60 μM), enalapril (an inhibitor of angiotensin converting enzyme, 150 μM), and cilnidipine (a voltage-dependent L-type Ca2+ channel blocker, 3 μM) significantly reduced the CA secretion evoked by ACh and angiotensin II (Ang II), respectively. In the simultaneous presence of ginsenoside-Rg3 (10 μM) along with four antihypertensive agents (olmesartan, enalapril, simvastatin or cilnidipine), the CA secretion evoked by ACh and Ang II was more strongly inhibited compared with that of each antihypertensive agent-treatment alone. Conclusions: Collectively, these results demonstrate that ginsenoside-Rg3 as well as antihypertensive agents can inhibit the CA secretion evoked by activation of both cholinergic and AT1 receptors from the perfused rat adrenal medulla. When this ginsenoside-Rg3 was used in combination with different antihypertensive agents, their effects of antihypertensive agents on reduction of adrenal CA secretion were significantly enhanced, which may also be clinically beneficial for the treatment of cardiovascular diseases.

INFLUENCE OF CHRONIC AT1 RECEPTOR BLOCKADE ON GASTRIC BARRIER FUNCTION IN MICE

Objective: The AT1 receptor blocker Telmisartan (TEL) is beneficial for the treatment of individuals suffering the Metabolic Syndrome as TEL reveals not only antihypertensive effects but also improves the glucose and lipid control and lowers body weight. As we have recently shown that TEL has impact on gut microbiota and brain barrier function we here investigated whether TEL influences gut barrier function which may participate in decrease of energy intake. Design and method: BL6 mice were fed with chow or high fat diet (45% fat) and were additionally treated with vehicle (CONchow or CONHFD) or TEL (8 mg/kg/d, 12 w, TELchow or TELHFD). Visceral and subcutaneous fat mass was quantified by MRI. Segments of large intestine were Canoy fixed to determine thickness of mucus by immunohistochemistry. PAS staining was performed for depicting goblet cells. The signalling pathway being involved in necroptosis was investigated by Western blots and qPCR. Results: Compared to chow feeding, gain in body weight increased by HFD (3.4±0.5 vs. 14.4g±1.2 g) and was reduced by TEL (1.7±0.5 vs. 2.8±0.5g). Visceral fat and liver mass was higher in CONHFD than CONchow but remained normal by TEL treatment. The mucus thickness was lower in CONHFD than CONchow. Independent on feeding, TEL additionally reduced mucus thickness. Numbers of goblet cells were not affected by HFD feeding and TEL treatment. RIP3 was particularly increased in TELHFD mice, while muc2 and ki67 almost remained unaffected by TEL. Conclusions: In contrast to our expectation, the antiobese effect of TEL was associated with a decrease in mucus thickness. Increase in RIP3 indicates that necroptosis may be enhanced by TEL. Further studies have to be performed to elucidate whether this detrimental TEL effect is true and be relevant for gastrointestinal function.

Intracellular Angiotensin II Stimulation of Sodium Transporter Expression in Proximal Tubule Cells via AT1 (AT1a) Receptor-Mediated, MAP Kinases ERK1/2- and NF-кB-Dependent Signaling Pathways.

The current prevailing paradigm in the renin-angiotensin system dictates that most, if not all, biological, physiological, and pathological responses to its most potent peptide, angiotensin II (Ang II), are mediated by extracellular Ang II activating its cell surface receptors. Whether intracellular (or intracrine) Ang II and its receptors are involved remains incompletely understood. The present study tested the hypothesis that extracellular Ang II is taken up by the proximal tubules of the kidney by an AT1 (AT1a) receptor-dependent mechanism and that overexpression of an intracellular Ang II fusion protein (ECFP/Ang II) in mouse proximal tubule cells (mPTC) stimulates the expression of Na+/H+ exchanger 3 (NHE3), Na+/HCO3- cotransporter, and sodium and glucose cotransporter 2 (Sglt2) by AT1a/MAPK/ERK1/2/NF-kB signaling pathways. mPCT cells derived from male wild-type and type 1a Ang II receptor-deficient mice (Agtr1a-/-) were transfected with an intracellular enhanced cyan fluorescent protein-tagged Ang II fusion protein, ECFP/Ang II, and treated without or with AT1 receptor blocker losartan, AT2 receptor blocker PD123319, MEK1/MEK2 inhibitor U0126, NF-кB inhibitor RO 106-9920, or p38 MAP kinase inhibitor SB202196, respectively. In wild-type mPCT cells, the expression of ECFP/Ang II significantly increased NHE3, Na+/HCO3-, and Sglt2 expression (p < 0.01). These responses were accompanied by >3-fold increases in the expression of phospho-ERK1/2 and the p65 subunit of NF-кB (p < 0.01). Losartan, U0126, or RO 106-9920 all significantly attenuated ECFP/Ang II-induced NHE3 and Na+/HCO3- expression (p < 0.01). Deletion of AT1 (AT1a) receptors in mPCT cells attenuated ECFP/Ang II-induced NHE3 and Na+/HCO3- expression (p < 0.01). Interestingly, the AT2 receptor blocker PD123319 also attenuated ECFP/Ang II-induced NHE3 and Na+/HCO3- expression (p < 0.01). These results suggest that, similar to extracellular Ang II, intracellular Ang II may also play an important role in Ang II receptor-mediated proximal tubule NHE3, Na+/HCO3-, and Sglt2 expression by activation of AT1a/MAPK/ERK1/2/NF-kB signaling pathways.

Cluster analysis of angiotensin biomarkers to identify antihypertensive drug treatment in population studies

BackgroundThe recent progress in molecular biology generates an increasing interest in investigating molecular biomarkers as markers of response to treatments. The present work is motivated by a study, where the objective was to explore the potential of the molecular biomarkers of renin-angiotensin-aldosterone system (RAAS) to identify the undertaken antihypertensive treatments in the general population. Population-based studies offer an opportunity to assess the effectiveness of treatments in real-world scenarios. However, lack of quality documentation, especially when electronic health record linkage is unavailable, leads to inaccurate reporting and classification bias.MethodWe present a machine learning clustering technique to determine the potential of measured RAAS biomarkers for the identification of undertaken treatments in the general population. The biomarkers were simultaneously determined through a novel mass-spectrometry analysis in 800 participants of the Cooperative Health Research In South Tyrol (CHRIS) study with documented antihypertensive treatments. We assessed the agreement, sensitivity and specificity of the resulting clusters against known treatment types. Through the lasso penalized regression, we identified clinical characteristics associated with the biomarkers, accounting for the effects of cluster and treatment classifications.ResultsWe identified three well-separated clusters: cluster 1 (n = 444) preferentially including individuals not receiving RAAS-targeting drugs; cluster 2 (n = 235) identifying angiotensin type 1 receptor blockers (ARB) users (weighted kappa κw = 74%; sensitivity = 73%; specificity = 83%); and cluster 3 (n = 121) well discriminating angiotensin-converting enzyme inhibitors (ACEi) users (κw = 81%; sensitivity = 55%; specificity = 90%). Individuals in clusters 2 and 3 had higher frequency of diabetes as well as higher fasting glucose and BMI levels. Age, sex and kidney function were strong predictors of the RAAS biomarkers independently of the cluster structure.ConclusionsUnsupervised clustering of angiotensin-based biomarkers is a viable technique to identify individuals on specific antihypertensive treatments, pointing to a potential application of the biomarkers as useful clinical diagnostic tools even outside of a controlled clinical setting.

High salt induces cognitive impairment via the interaction of the angiotensin II-AT1 and prostaglandin E2-EP1 systems.

High salt (HS) intake has been associated with hypertension and cognitive impairment. It is well known that the angiotensin II (Ang II)-AT1 receptor and prostaglandin E2 (PGE2)-EP1 receptor systems are involved in hypertension and neurotoxicity. However, the involvement of these systems in HS-mediated hypertension and emotional and cognitive impairments remains unclear. Mice were loaded with HS solution (2% NaCl drinking water) for 12 weeks, and blood pressure was monitored. Subsequently, effects of HS intake on emotional and cognitive function and tau phosphorylation in the prefrontal cortex (PFC) and hippocampus (HIP) were investigated. The involvement of Ang II-AT1 and PGE2-EP1 systems in HS-induced hypertension and neuronal and behavioural impairments was examined by treatment with losartan, an AT1 receptor blocker (ARB), or EP1 gene knockout. We demonstrate that hypertension and impaired social behaviour and object recognition memory following HS intake may be associated with tau hyperphosphorylation, decreased phosphorylation of Ca2+ /calmodulin-dependent protein kinase II (CaMKII), and postsynaptic density protein 95 (PSD95) expression in the PFC and HIP of mice. These changes were blocked by pharmacological treatment with losartan or EP1 receptor gene knockout. Our findings suggest that the interaction of Ang II-AT1 receptor and PGE2-EP1 receptor systems could be novel therapeutic targets for hypertension-induced cognitive impairment.

Repurposing of RAS-Pathway Mediated Drugs for Intestinal Inflammation Related Diseases for Treating SARS-CoV-2 Infection

Severe Acute Respiratory Syndrome coronavirus 2 (SARS-CoV-2) is an emerging zoonotic virus, which causes Coronavirus Disease 2019 (COVID-19). Entry of coronaviruses into the cell depends on binding of the viral spike (S) proteins to cellular receptors Angiotensin-converting enzyme 2 (ACE2). The virus-mediated reduction of ACE2/Ang1-7 causes flooding of inflammatory cytokines. A similar scenario of hyper immunologic reaction has been witnessed in the context of Intestinal Inflammatory Diseases (IIDs) with the deregulation of ACE2. This review summarizes several IIDs that lead to such susceptible conditions. It discusses suitable mechanisms of how ACE2, being a crucial regulator of the Renin-Angiotensin System (RAS) signaling pathway, can affect the physiology of intestine as well as lungs, the primary site of SARS-CoV-2 infection. ACE2, as a SARS-CoV-2 receptor, establishes a critical link between COVID-19 and IIDs. Intercessional studies targeting the RAS signaling pathway in patients may provide a novel strategy for addressing the COVID-19 crisis. Hence, the modulation of these key RAS pathway members can be beneficial in both instances. However, it's difficult to say how beneficial are the ACE inhibitors (ACEI)/ Angiotensin II type-1 receptor blockers (ARBs) during COVID-19. As a result, much more research is needed to better understand the relationship between the RAS and SARS-CoV-2 infection.