ABSTRACT
Myocardial fibrosis following acute myocardial infarction (AMI) seriously affects the prognosis and survival rate of patients. This study explores the role and regulation mechanism of storax, a commonly used traditional Chinese medicine for treatment of cardiovascular diseases, on myocardial fibrosis and cardiac function. The AMI rat model was established by subcutaneous injection of Isoproterenol hydrochloride (ISO). Storax (0.1, 0.2, 0.4 g/kg) was administered by gavage once/d for 7 days. Electrocardiogram, echocardiography, hemodynamic and cardiac enzyme in AMI rats were measured. HE, Masson, immunofluorescence and TUNEL staining were used to observe the degree of pathological damage, fibrosis and cardiomyocyte apoptosis in myocardial tissue, respectively. Expression of AT1R, CARP and their downstream related apoptotic proteins were detected by WB. The results demonstrated that storax could significantly improve cardiac electrophysiology and function, decrease serum cardiac enzyme activity, reduce type I and III collagen contents to improve fibrosis and alleviate myocardial pathological damage and cardiomyocyte apoptosis. It also found that storax can significantly down-regulate expression of AT1R, Ankrd1, P53, P-p53 (ser 15), Bax and cleaved Caspase-3 and up-regulate expression of Mdm2 and Bcl-2. Taken together, these findings indicated that storax effectively protected cardiomyocytes against myocardial fibrosis and cardiac dysfunction by inhibiting the AT1R-Ankrd1-P53 signaling pathway.
Subject(s)
Drugs, Chinese Herbal , Myocardial Infarction , Animals , Rats , Apoptosis , Disease Models, Animal , Drugs, Chinese Herbal/pharmacology , Drugs, Chinese Herbal/therapeutic use , Fibrosis , Muscle Proteins/drug effects , Muscle Proteins/metabolism , Myocardial Infarction/complications , Myocardial Infarction/drug therapy , Myocardial Infarction/metabolism , Myocardium/metabolism , Myocytes, Cardiac/metabolism , Nuclear Proteins/drug effects , Nuclear Proteins/metabolism , Receptor, Angiotensin, Type 1/drug effects , Receptor, Angiotensin, Type 1/metabolism , Repressor Proteins/drug effects , Repressor Proteins/metabolism , Signal Transduction , Tumor Suppressor Protein p53/drug effects , Tumor Suppressor Protein p53/metabolismABSTRACT
Blood pressure is controlled by endocrine, autonomic, and behavioral responses that maintain blood volume and perfusion pressure at levels optimal for survival. Although it is clear that central angiotensin type 1a receptors (AT1aR; encoded by the Agtr1a gene) influence these processes, the neuronal circuits mediating these effects are incompletely understood. The present studies characterize the structure and function of AT1aR neurons in the lamina terminalis (containing the median preoptic nucleus and organum vasculosum of the lamina terminalis), thereby evaluating their roles in blood pressure control. Using male Agtr1a-Cre mice, neuroanatomical studies reveal that AT1aR neurons in the area are largely glutamatergic and send projections to the paraventricular nucleus of the hypothalamus (PVN) that appear to synapse onto vasopressin-synthesizing neurons. To evaluate the functionality of these lamina terminalis AT1aR neurons, we virally delivered light-sensitive opsins and then optogenetically excited or inhibited the neurons while evaluating cardiovascular parameters or fluid intake. Optogenetic excitation robustly elevated blood pressure, water intake, and sodium intake, while optogenetic inhibition produced the opposite effects. Intriguingly, optogenetic excitation of these AT1aR neurons of the lamina terminalis also resulted in Fos induction in vasopressin neurons within the PVN and supraoptic nucleus. Further, within the PVN, selective optogenetic stimulation of afferents that arise from these lamina terminalis AT1aR neurons induced glutamate release onto magnocellular neurons and was sufficient to increase blood pressure. These cardiovascular effects were attenuated by systemic pretreatment with a vasopressin-1a-receptor antagonist. Collectively, these data indicate that excitation of lamina terminalis AT1aR neurons induces neuroendocrine and behavioral responses that increase blood pressure.SIGNIFICANCE STATEMENT Hypertension is a widespread health problem and risk factor for cardiovascular disease. Although treatments exist, a substantial percentage of patients suffer from "drug-resistant" hypertension, a condition associated with increased activation of brain angiotensin receptors, enhanced sympathetic nervous system activity, and elevated vasopressin levels. The present study highlights a role for angiotensin Type 1a receptor expressing neurons located within the lamina terminalis in regulating endocrine and behavioral responses that are involved in maintaining cardiovascular homeostasis. More specifically, data presented here reveal functional excitatory connections between angiotensin-sensitive neurons in the lamina terminals and vasopressin neurons in the paraventricular nucleus of the hypothalamus, and further indicate that activation of this circuit raises blood pressure. These neurons may be a promising target for antihypertensive therapeutics.
Subject(s)
Angiotensins/pharmacology , Arginine Vasopressin/metabolism , Blood Pressure/drug effects , Hypothalamus/drug effects , Neural Pathways/drug effects , Paraventricular Hypothalamic Nucleus/drug effects , Vasoconstrictor Agents/pharmacology , Animals , Basal Nucleus of Meynert/drug effects , Basal Nucleus of Meynert/metabolism , Drinking/drug effects , Genes, fos/drug effects , Glutamic Acid/physiology , Male , Mice , Mice, Inbred C57BL , Mice, Knockout , Optogenetics , Receptor, Angiotensin, Type 1/drug effects , Receptors, Vasopressin/drug effects , Sodium, DietaryABSTRACT
AIM: Vitamin D plays an important role in water and salt homeostasis. The aim of our study was to investigate the underlying relationship of Vitamin D and Aquaporins (AQP). METHODS: The behaviors of 1α (OH)-ase knockout mice and wild type mice were observed before analysis. The ICR mice were treated with vehicle or paricalcitol, a vitamin D analogue, followed by animals receiving a standard diet and free access to drinking water either with aliskiren (renin blocker; 37.5 mg aliskiren in 100 ml water), or telmisartan (a angiotensin II type I receptor blocker; 40 mg telmisartan in 100 ml water) a week before study. The expressions of AQP-1, AQP-4, and renin in mice kidneys were detected by western bolting, immunohistochemistry, and immunofluorescence. RESULTS: Diuresis and polydipsia were observed in 1α (OH)-ase knockout mice, and a decreased water intake and urine output in ICR mice was observed after paricalcitol treatment. Compared with wild type, the AQP-1 expressions were increased in renal papilla and AQP-4 expressions were decreased in renal proximal tubule of 1α(OH) ase knockout mice. In addition, AQP-1 was decreased in renal papilla and AQP-4 expressions were increased in proximal tubule by suppressing renin activity or supplement of Vitamin D analogue. After injecting renin into the lateral ventricle of the 1α(OH)ase knockout mice, the renin expression level was decreased in the kidney, followed by the decrease of AQP-1 in renal papilla and increase of AQP-4 in proximal tubule. CONCLUSIONS: Overall, Vitamin D and renin inhibitors have synergistic effects in regulating water channels in mice kidneys.
Subject(s)
Aquaporin 1/genetics , Aquaporin 4/genetics , Renin/genetics , Vitamin D/analogs & derivatives , Vitamin D/metabolism , Amides/administration & dosage , Angiotensin II Type 1 Receptor Blockers/administration & dosage , Animals , Fumarates/administration & dosage , Gene Expression Regulation/drug effects , Humans , Kidney/drug effects , Kidney/metabolism , Kidney/pathology , Kidney Tubules, Proximal/drug effects , Kidney Tubules, Proximal/metabolism , Mice , Mice, Knockout , Receptor, Angiotensin, Type 1/drug effects , Renin/administration & dosage , Renin/antagonists & inhibitors , Renin-Angiotensin System/drug effects , Telmisartan/administration & dosage , Vitamin D/genetics , Water/chemistryABSTRACT
Carthamus tinctorius L. (CT) is widely used in Asian countries as a beverage and in folk medicine. The effects of CT extract on hemodynamics, vascular remodeling, the renin-angiotensin system (RAS) and oxidative stress in the two-kidney, one clip (2K-1C) hypertensive rat model were investigated. Renovascular hypertension was induced in male Sprague-Dawley rats and were treated with CT extract (500mg/kg/day) or captopril (5mg/kg/day) or vehicle for four weeks. CT extract or captopril reduced blood pressure, hindlimb vascular resistance, and increased hindlimb blood flow in 2K-1C hypertensive rats (p<0.05). Increases in aortic wall thickness, cross-sectional area and collagen deposition in 2K-1C rats were alleviated with CT extract or captopril treatment (p<0.05). CT extract or captopril suppressed RAS activation, including elevated serum ACE activity, and plasma Ang II level and up-regulated aortic AT1R protein expression in 2K-1C rats (p<0.05). Furthermore, CT extract or captopril reduced vascular superoxide production, aortic NADPH oxidase subunit gp91phox expression and increased plasma nitric oxide metabolite levels in 2K-1C rats (p<0.05). These findings suggest that CT extract ameliorated hemodynamic alteration and vascular remodeling in 2K-1C hypertensive rats. Possible mechanisms may involve RAS inhibitor effects and potent antioxidant activity.
Subject(s)
Carthamus tinctorius/chemistry , Hemodynamics/drug effects , Hypertension, Renovascular/drug therapy , Plant Extracts/therapeutic use , Renin-Angiotensin System/drug effects , Signal Transduction/drug effects , Angiotensin II/drug effects , Animals , Blood Pressure/drug effects , Hindlimb/blood supply , Hindlimb/drug effects , Hypertension, Renovascular/physiopathology , Male , NADPH Oxidases/drug effects , Nitric Oxide Synthase/metabolism , Oxidative Stress/drug effects , Rats , Rats, Sprague-Dawley , Receptor, Angiotensin, Type 1/drug effects , Regional Blood Flow/drug effects , Vascular Resistance/drug effectsABSTRACT
A role for endothelium-derived constricting factors (EDCF), and the angiotensin II type 1 receptor (AT1R) pathway, in the vascular impairment found in the rat Freund's complete adjuvant (FCA)-model of rheumatoid arthritis (RA) was examined. FCA arthritis was induced in rats±losartan. Vehicle-treated rats served as controls. Knee-joint swelling and red blood cell (RBC) aggregation were measured as indicators of inflammation and endothelium reactivity assessed by response to acetylcholine (ACh) on aortic rings. Results show that knee-joint swelling and RBC aggregation were elevated in the FCA+vehicle group and restored to control levels in the FCA+losartan-treated animals. ACh-induced relaxation of aortic rings taken from FCA+vehicle animals was significantly impaired compared to vehicle-controls and this vasoreactivity was restored to control levels in the FCA+losartan-treated group. Further examination of aorta from the FCA+vehicle animals revealed an EDCF that was reliant on cyclooxygenase-2 (but not cyclooxygenase-1), generation of superoxide anion generation (but not hydrogen peroxide) and activation of thromboxane-prostanoid receptor. Losartan administration in vivo or ex vivo (to aortic rings) prevented the generation of the EDCF. In summary, this is the first evidence of an EDCF in a model of RA and identifies this mechanism as potentially significant in the cardiovascular disorder associated with the disease.
Subject(s)
Aorta, Thoracic/metabolism , Arthritis, Experimental/metabolism , Endothelium, Vascular/metabolism , Receptor, Angiotensin, Type 1/metabolism , Vasoconstriction , Angiotensin II Type 1 Receptor Blockers/pharmacology , Animals , Antirheumatic Agents/pharmacology , Aorta, Thoracic/drug effects , Aorta, Thoracic/physiopathology , Arthritis, Experimental/chemically induced , Arthritis, Experimental/drug therapy , Arthritis, Experimental/physiopathology , Cyclooxygenase 2/metabolism , Endothelium, Vascular/drug effects , Endothelium, Vascular/physiopathology , Freund's Adjuvant , Losartan/pharmacology , Male , Rats, Sprague-Dawley , Receptor, Angiotensin, Type 1/drug effects , Receptors, Thromboxane/metabolism , Signal Transduction , Superoxides/metabolism , Vasoconstriction/drug effects , Vasodilator Agents/pharmacologyABSTRACT
This study examined the effect of Carthamus tinctorius (CT) extract plus captopril treatment on blood pressure, vascular function, nitric oxide (NO) bioavailability, oxidative stress and renin-angiotensin system (RAS) in N(ω)-Nitro-l-arginine methyl ester (l-NAME)-induced hypertension. Rats were treated with l-NAME (40 mg/kg/day) for five weeks and given CT extract (75 or 150 or 300 or 500 mg/kg/day): captopril (5 mg/kg/day) or CT extract (300 mg/kg/day) plus captopril (5 mg/kg/day) for two consecutive weeks. CT extract reduced blood pressure dose-dependently, and the most effective dose was 300 mg/kg/day. l-NAME-induced hypertensive rats showed abnormalities including high blood pressure, high vascular resistance, impairment of acetylcholine-induced vasorelaxation in isolated aortic rings and mesenteric vascular beds, increased vascular superoxide production and plasma malondialdehyde levels, downregulation of eNOS, low level of plasma nitric oxide metabolites, upregulation of angiotensin II type 1 receptor and increased plasma angiotensin II. These abnormalities were alleviated by treatment with either CT extract or captopril. Combination treatment of CT extract and captopril normalized all the abnormalities found in hypertensive rats except endothelial dysfunction. These data indicate that there are synergistic antihypertensive effects of CT extract and captopril. These effects are likely mediated by their anti-oxidative properties and their inhibition of RAS.
Subject(s)
Angiotensin II Type 1 Receptor Blockers/pharmacology , Antihypertensive Agents/pharmacology , Blood Pressure/drug effects , Captopril/pharmacology , Carthamus tinctorius/chemistry , Hypertension/drug therapy , NG-Nitroarginine Methyl Ester , Nitric Oxide Synthase Type III/metabolism , Plant Extracts/pharmacology , Receptor, Angiotensin, Type 1/drug effects , Animals , Antihypertensive Agents/isolation & purification , Disease Models, Animal , Dose-Response Relationship, Drug , Drug Synergism , Hypertension/chemically induced , Hypertension/enzymology , Hypertension/physiopathology , Male , Nitric Oxide/metabolism , Oxidative Stress/drug effects , Phytotherapy , Plant Extracts/isolation & purification , Plants, Medicinal , Rats, Sprague-Dawley , Receptor, Angiotensin, Type 1/metabolism , Renin-Angiotensin System/drug effects , Time Factors , Vasodilation/drug effectsABSTRACT
It is of great therapeutic significance that disordered function of the vascular endothelium which supply the affected ocular structures plays a major role in the pathogenesis and development of age-related macular degeneration. Chronic inflammation is closely linked to diseases associated with endothelial dysfunction, and age-related macular degeneration is accompanied by a general inflammatory response. According to current concept, age-related macular degeneration is a local manifestation of systemic vascular disease. This recognition could have therapeutic implications because restoration of endothelial dysfunction can restabilize the condition of chronic vascular disease including age-related macular degeneration as well. Restoration of endothelial dysfunction by pharmaacological or non pharmacological interventions may prevent the development or improve endothelial dysfunction, which result in prevention or improvement of age related macular degeneration as well. Medicines including inhibitors of the renin-angiotensin system (converting enzyme inhibitors, angiotensin-receptor blockers and renin inhibitors), statins, acetylsalicylic acid, trimetazidin, third generation beta-blockers, peroxisome proliferator-activated receptor gamma agonists, folate, vitamin D, melatonin, advanced glycation end-product crosslink breaker alagebrium, endothelin-receptor antagonist bosentan, coenzyme Q10; "causal" antioxidant vitamins, N-acetyl-cysteine, resveratrol, L-arginine, serotonin receptor agonists, tumor necrosis factor-alpha blockers, specific inhibitor of the complement alternative pathway, curcumin and doxycyclin all have beneficial effects on endothelial dysfunction. Restoration of endothelial dysfunction can restabilize chronic vascular disease including age-related macular degeneration as well. Considering that the human vascular system is consubstantial, medicines listed above should be given to patients (1) who have no macular degeneration but have risk factors for the disease and are older than 50 years; (2) who have been diagnosed with unilateral age-related macular degeneration in order to prevent damage of the contralateral eye; (3) who have bilateral age-related macular degeneration in order to avert deterioration and in the hope of a potential improvement. However, randomised prospective clinical trials are still needed to elucidate the potential role of these drug treatments in the prevention and treatment of age-related macular degeneration.
Subject(s)
Dietary Supplements , Macular Degeneration/drug therapy , Macular Degeneration/metabolism , Renin-Angiotensin System/drug effects , Aged , Aged, 80 and over , Angiotensin-Converting Enzyme Inhibitors/therapeutic use , Antioxidants/administration & dosage , Aspirin/administration & dosage , Aspirin/analogs & derivatives , Bosentan , Docosahexaenoic Acids/administration & dosage , Eicosapentaenoic Acid/administration & dosage , Folic Acid/administration & dosage , Humans , Hydroxymethylglutaryl-CoA Reductase Inhibitors/administration & dosage , Infliximab/administration & dosage , Lutein/administration & dosage , Melatonin/administration & dosage , PPAR gamma/agonists , Receptor, Angiotensin, Type 1/drug effects , Renin/antagonists & inhibitors , Resveratrol , Stilbenes/administration & dosage , Sulfonamides/administration & dosage , Trimetazidine/administration & dosage , Tumor Necrosis Factor-alpha/antagonists & inhibitors , Ubiquinone/administration & dosage , Ubiquinone/analogs & derivatives , Vitamin D/administration & dosage , Xanthophylls/administration & dosageABSTRACT
The existence of a local renin-angiotensin system (RAS) in neurons was first postulated 40 years ago. Further studies indicated intraneuronal generation of ANG II. However, the function and signaling mechanisms of intraneuronal ANG II remained elusive. Since ANG II type 1 receptor (AT1R) is the major type of receptor mediating the effects of ANG II, we used intracellular microinjection and concurrent Ca(2+) and voltage imaging to examine the functionality of intracellular AT1R in neurons. We show that intracellular administration of ANG II produces a dose-dependent elevation of cytosolic Ca(2+) concentration ([Ca(2+)]i) in hypothalamic neurons that is sensitive to AT1R antagonism. Endolysosomal, but not Golgi apparatus, disruption prevents the effect of microinjected ANG II on [Ca(2+)]i. Additionally, the ANG II-induced Ca(2+) response is dependent on microautophagy and sensitive to inhibition of PLC or antagonism of inositol 1,4,5-trisphosphate receptors. Furthermore, intracellular application of ANG II produces AT1R-mediated depolarization of hypothalamic neurons, which is dependent on [Ca(2+)]i increase and on cation influx via transient receptor potential canonical channels. In summary, we provide evidence that intracellular ANG II activates endolysosomal AT1Rs in hypothalamic neurons. Our results point to the functionality of a novel intraneuronal angiotensinergic pathway, extending the current understanding of intracrine ANG II signaling.
Subject(s)
Angiotensin II/metabolism , Neurons/physiology , Signal Transduction/physiology , Angiotensin II/administration & dosage , Angiotensin II/pharmacology , Angiotensin II Type 1 Receptor Blockers/administration & dosage , Angiotensin II Type 1 Receptor Blockers/pharmacology , Animals , Animals, Newborn , Calcium/metabolism , Cell Line, Tumor , Dose-Response Relationship, Drug , Female , Gene Expression Regulation , Humans , Hypothalamus/cytology , Inositol 1,4,5-Trisphosphate/metabolism , Inositol 1,4,5-Trisphosphate Receptors/genetics , Inositol 1,4,5-Trisphosphate Receptors/metabolism , Male , Microinjections , Neurons/drug effects , Rats , Rats, Sprague-Dawley , Receptor, Angiotensin, Type 1/drug effects , Receptor, Angiotensin, Type 1/metabolismABSTRACT
Angiotensin (Ang) II and its AT1 receptors have been implicated in the pathogenesis of rheumatoid arthritis. Activation of the counter-regulatory Ang-(1-7)-Mas receptor axis may contribute to some of the effects of AT1 receptor blockers (ARBs). In this study, we have used losartan, an ARB, to investigate the role of and the mechanisms by which AT1 receptors participated in two experimental models of arthritis: antigen-induced arthritis (AIA) in mice and adjuvant-induced arthritis (AdIA) in rats. Treatment with losartan decreased neutrophil recruitment, hypernociception and the production of TNF-α, IL-1ß and chemokine (C-X-C motif) ligand 1 in mice subjected to AIA. Histopathological analysis showed significant reduction of tissue injury and inflammation and decreased proteoglycan loss. In addition to decreasing cytokine production, losartan directly reduced leukocyte rolling and adhesion. Anti-inflammatory effects of losartan were not associated to Mas receptor activation and/or Ang-(1-7) production. Anti-inflammatory effects were reproduced in rats subjected to AdIA. This study shows that ARBs have potent anti-inflammatory effects in animal models of arthritis. Mechanistically, reduction of leukocyte accumulation and of joint damage was associated with local inhibition of cytokine production and direct inhibition of leukocyte-endothelium interactions. The anti-inflammatory actions of losartan were accompanied by functional improvement of the joint, as seen by reduced joint hypernociception. These findings support the use of ARBs for the treatment of human arthritis and provide potential mechanisms for the anti-inflammatory actions of these compounds.
Subject(s)
Angiotensin II Type 1 Receptor Blockers/pharmacology , Anti-Inflammatory Agents, Non-Steroidal/pharmacology , Arthritis, Experimental/drug therapy , Losartan/pharmacology , Receptor, Angiotensin, Type 1/drug effects , Angiotensin I/biosynthesis , Animals , Arthritis, Rheumatoid/drug therapy , Cell Adhesion/drug effects , Chemokine CXCL1/biosynthesis , Disease Models, Animal , Female , Hyperalgesia/drug therapy , Inflammation/drug therapy , Interleukin-1beta/biosynthesis , Leukocyte Rolling/drug effects , Male , Mice , Mice, Inbred C57BL , Neutrophil Infiltration/drug effects , Peptide Fragments/biosynthesis , Proto-Oncogene Mas , Proto-Oncogene Proteins/metabolism , Rats , Rats, Sprague-Dawley , Receptor, Angiotensin, Type 1/metabolism , Receptors, G-Protein-Coupled/metabolism , Tumor Necrosis Factor-alpha/biosynthesisABSTRACT
AIMS: Rheumatoid arthritis (RA) is associated with high cardiovascular mortality. Impaired endothelial cell (EC) function and elevated angiotensin II levels may be central to the link between vascular dysfunction and RA. Here we investigated the action of angiotensin type 1 receptor (AT1R) blockade on endothelium-dependent relaxation of the isolated saphenous artery in a rat model of monoarthritis. MAIN METHODS: Adjuvant arthritis was induced in rats with and without prophylactic losartan (AT1R antagonist) treatment. Vehicle-treated rats were used as controls. Wire myography was employed to investigate EC function of isolated rings of saphenous artery. KEY FINDINGS: EC-dependent relaxation in arteries from non-inflamed control rats was mediated by both nitric oxide (NO) and endothelium-derived hyperpolarising factor (EDHF) with the EDHF response dependent principally on functional myoendothelial gap junctions. While NO-dependent relaxation remained unaffected, the EDHF-mediated response was abolished in arteries from arthritic rats (P<0.001), however, substantial protection (approximately 50%) of the EDHF-relaxation was found in arthritic rats treated with losartan (P<0.01). Thus, the attenuated EDHF response found in the saphenous artery of arthritic rats was significantly reversed by AT1R blockade. SIGNIFICANCE: These results suggest a key role for the angiotensin system in the EC dysfunction found in chronic joint inflammation and highlights AT1R as a potential therapeutic target to redress the vascular impairment and mortality associated with RA.
Subject(s)
Angiotensin II Type 1 Receptor Blockers/pharmacology , Arthritis, Experimental/physiopathology , Endothelium-Dependent Relaxing Factors/physiology , Muscle, Smooth, Vascular/drug effects , Receptor, Angiotensin, Type 1/drug effects , Animals , Arteries/drug effects , Arteries/physiopathology , Aspirin/pharmacology , Charybdotoxin/pharmacology , Disease Models, Animal , Dose-Response Relationship, Drug , Female , Losartan/pharmacology , Muscle Relaxation/drug effects , Muscle Relaxation/physiology , Muscle, Smooth, Vascular/physiopathology , Myography , NG-Nitroarginine Methyl Ester/pharmacology , Rats , Rats, Sprague-Dawley , Receptor, Angiotensin, Type 1/physiologyABSTRACT
Renin-angiotensin system blockade improves glucose intolerance and insulin resistance, which contribute to the development of metabolic syndrome. However, the contribution of impaired insulin secretion to the pathogenesis of metabolic syndrome is not well defined. To assess the contributions of angiotensin receptor type 1 (AT1) activation and high glucose intake on pancreatic function and their effects on insulin signaling in skeletal muscle and adipose tissue, an oral glucose tolerance test (oGTT) was performed in five groups (n = 10/group) of rats: 1) lean strain-control 2) obese Otsuka Long-Evans Tokushima Fatty (OLETF), 3) OLETF + angiotensin receptor blocker (ARB; 10 mg/kg · d olmesartan for 6 wk; OLETF ARB), 4) OLETF + 5% glucose water (HG) for 6 wk (OLETF HG), and 5) OLETF + HG + ARB (OLETF HG/ARB). The glucose response to the oGTT increased 58% in OLETF compared with lean-strain control, whereas glucose supplementation increased it an additional 26%. Blockade of angiotensin receptor reduced the oGTT response 19% in the ARB-treated groups and increased pancreatic insulin secretion 64 and 113% in OLETF ARB and OLETF HG/ARB, respectively. ARB treatment in OLETF ARB and OLETF HG/ARB did not have an effect on insulin signaling proteins in skeletal muscle; however, it reduced pancreatic AT1 protein expression 20 and 27%, increased pancreatic glucagon-like peptide-1 (GLP-1) receptor protein expression 41 and 88%, respectively, and increased fasting plasma GLP-1 approximately 2.5-fold in OLETF ARB. The results suggest that improvement of glucose intolerance is independent of an improvement in muscle insulin signaling, but rather by improved glucose-stimulated insulin secretion associated with decreased pancreatic AT1 activation and increased GLP-1 signaling.
Subject(s)
Angiotensin II Type 1 Receptor Blockers/pharmacology , Glucose Intolerance/prevention & control , Glucose/pharmacology , Insulin/metabolism , Metabolic Syndrome/metabolism , Pancreas/metabolism , Receptor, Angiotensin, Type 1/drug effects , Adipose Tissue/drug effects , Adipose Tissue/metabolism , Animals , Blood Pressure/drug effects , Dietary Supplements , Disease Models, Animal , Glucagon-Like Peptide-1 Receptor , Glucose/administration & dosage , Glucose Intolerance/metabolism , Leptin/blood , Male , Muscle, Skeletal/drug effects , Muscle, Skeletal/metabolism , Rats , Rats, Inbred OLETF , Rats, Inbred Strains , Receptor, Angiotensin, Type 1/metabolism , Receptors, Glucagon/metabolism , Triglycerides/bloodABSTRACT
AIMS: We tested the hypothesis that candesartan, an angiotensin II (AII) type 1 receptor antagonist, would restore the depressed phosphatidylinositol 3 (PI3) kinase-dependent Akt phosphorylation, an essential signal to induce heat-shock protein 72 (Hsp72) in response to hyperthermia, in Otsuka Long-Evans Tokushima fatty (OLETF) rats. METHODS AND RESULTS: At 14 weeks of age, male OLETF rats and Long-Evans Tokushima Otsuka (LETO) rats were treated with candesartan (0.25 mg/kg/day) for 2 weeks. Thereafter, hyperthermia (43°C for 20 min) was applied. We observed the following: (i) Candesartan did not improve insulin sensitivity in OLETF rats. (ii) Candesartan restored depressed PI3 kinase-dependent Akt phosphorylation and Hsp72 expression in OLETF rat hearts. (iii) Cardiac ventricular tissue contents of AII were greater in OLETF rats, which were suppressed by candesartan. (iv) Cardiac levels of phosphatase and tensin homologue deleted on chromosome 10 (PTEN) phosphorylation were greater in OLETF rats, which were suppressed by candesartan. In cultured cardiomyocytes, application of AII induced PTEN phosphorylation, which was suppressed by candesartan. (v) In high-fat diet insulin-resistant rats, similar results were observed with respect to Hsp72 expression, Akt phosphorylation and PTEN phosphorylation. (vi) In isolated, perfused heart experiments, reperfusion-induced cardiac functional recovery was suppressed in OLETF rat hearts, which was improved by candesartan. CONCLUSION: Our results suggest that the depression of PI3 kinase-dependent Akt activation in response to hyperthermia in OLETF rats can be restored by candesartan. Substantial activation of the renin-angiotensin system, represented by increased myocardial AII content and subsequent PTEN phosphorylation, may underlie the pathogenesis which is ameliorated by candesartan.
Subject(s)
Angiotensin II Type 1 Receptor Blockers/pharmacology , Benzimidazoles/pharmacology , HSP72 Heat-Shock Proteins/metabolism , Hyperthermia, Induced , Insulin Resistance/genetics , Myocardial Reperfusion Injury/prevention & control , Myocardium/metabolism , Tetrazoles/pharmacology , Angiotensin II/metabolism , Animals , Animals, Newborn , Biphenyl Compounds , Blood Pressure , Cells, Cultured , Dietary Fats/administration & dosage , Disease Models, Animal , Genetic Predisposition to Disease , Glucose Tolerance Test , Glucose Transporter Type 4/metabolism , Heredity , Male , Myocardial Reperfusion Injury/genetics , Myocardial Reperfusion Injury/metabolism , Myocardial Reperfusion Injury/physiopathology , PTEN Phosphohydrolase/metabolism , Phosphatidylinositol 3-Kinase/metabolism , Phosphorylation , Proto-Oncogene Proteins c-akt/metabolism , Rats , Rats, Inbred OLETF , Rats, Long-Evans , Rats, Sprague-Dawley , Receptor, Angiotensin, Type 1/drug effects , Receptor, Angiotensin, Type 1/metabolism , Receptor, Angiotensin, Type 2/metabolism , Recovery of Function , Time Factors , Ventricular Function, Left , Ventricular PressureABSTRACT
BACKGROUND: Saponosides (horse chestnut seed extract, escin) and flavonoids (diosmin, Daflon 500, Servier, France) exhibit venotonic properties that have been utilized in treatment of varicose veins. However, the cellular mechanisms underlying the venotonic properties of escin and diosmin are unclear. Because Ca(2+) is a major regulator of venous smooth muscle (VSM) function, we tested the hypothesis that escin and diosmin promote Ca(2+)-dependent venous contraction. METHODS: Rings of inferior vena cava (IVC) from male rats were suspended in a tissue bath for measurement of isometric contraction. Following control contraction to 96 mM KCl, the effects of escin and diosmin (10(-10) to 10(-4) M) on vein contraction were measured. To test the role of intracellular Ca(2+) release, the vein response to escin and diosmin was measured in Ca(2+)-free (2mM EGTA) Krebs. To test for Ca(2+)-dependent effects, IVC segments were pretreated with escin or diosmin (10(-4) M) in 0 Ca(2+) Krebs, then extracellular CaCl(2) (0.1, 0.3, 0.6, 1, 2.5 mM) was added and the [Ca(2+)](e)-contraction relationship was constructed. To test for synergistic effects of diosmin, IVC segments were pretreated with diosmin (10(-4) M), then stimulated with KCl (16-96 mM) or escin (10(-10) to 10(-4) M) and vein contraction was measured. Contraction data were presented as mg/mg tissue (means ± SEM). RESULTS: In IVC segments incubated in normal Krebs (2.5 mM Ca(2+)), escin caused concentration-dependent contraction (max 104.3 ± 19.6 at 10(-4) M). Escin-induced contraction was not a rigor state, because after washing with Krebs, the veins returned to a relaxed state. In Ca(2+)-free Krebs, there was essentially no contraction to escin. In escin-treated veins incubated in 0 Ca(2+) Krebs, stepwise addition of extracellular CaCl(2) caused corresponding increases in contraction (max 80.0 ± 11.1 at 2.5 mM). In the absence of escin, the α-adrenergic agonist phenylephrine (PHE, 10(-5) M), angiotensin II (AngII, 10(-6) M), and membrane depolarization by KCl (96 mM) caused significant contraction (122.5 ± 45.1, 114.2 ± 12.2 and 221.7 ± 35.4, respectively). In IVC segments pretreated with escin (10(-4) M), the contractile response to PHE (9.7 ± 2.6), AngII (36.0 ± 9.1), and KCl (82.3 ± 10.2) was significantly reduced. Diosmin (10(-4) M) caused small contractions in normal Krebs (11.7 ± 1.9) and Ca(2+)-free Krebs (4.2 ± 2.2). In diosmin-treated veins incubated in 0 Ca(2+) Krebs, addition of extracellular CaCl(2) caused minimal contraction. Diosmin did not enhance the IVC contraction to PHE, AngII, or escin, but enhanced the contractile response to KCl (24-51 mM). CONCLUSION: In rat IVC, escin induces extracellular Ca(2+)-dependent contraction, but disrupts α-adrenergic and AT(1)R receptor-mediated pathways and depolarization-induced contraction. The initial venotonic benefits of escin may be offset by disruption of vein response to endogenous venoconstrictors, limiting its long-term therapeutic benefits in varicose veins. Diosmin does not cause venous contraction or potentiate the venotonic effects of endogenous venoconstrictors or escin ex vivo, and its use as venotonic may need to be further evaluated.
Subject(s)
Calcium Signaling/drug effects , Escin/pharmacology , Muscle, Smooth, Vascular/drug effects , Varicose Veins/drug therapy , Vasoconstriction/drug effects , Vasoconstrictor Agents/pharmacology , Vena Cava, Inferior/drug effects , Adrenergic alpha-1 Receptor Agonists/pharmacology , Angiotensin II/pharmacology , Animals , Diosmin/pharmacology , Dose-Response Relationship, Drug , In Vitro Techniques , Male , Membrane Potentials , Muscle, Smooth, Vascular/metabolism , Phenylephrine/pharmacology , Potassium Chloride/pharmacology , Rats , Rats, Sprague-Dawley , Receptor, Angiotensin, Type 1/drug effects , Receptor, Angiotensin, Type 1/metabolism , Receptors, Adrenergic, alpha-1/drug effects , Receptors, Adrenergic, alpha-1/metabolism , Varicose Veins/metabolism , Varicose Veins/physiopathology , Vena Cava, Inferior/metabolismABSTRACT
Calcilytics, antagonists of calcium receptor, decrease sensitivity of this receptor to plasma calcium concentration and increase parathyroid hormone (PTH) secretion. Moreover, it was recently indicated that calcilytic NPS 2143 induces hypertension in rats. This study tested whether the increase of mean arterial blood pressure (MAP) induced by NPS 2143 administration is mediated by calcium channel and angiotensin II type 1 (AT1) receptor activity. Wistar rats were anaesthesized with Thiopental i.p. and infused i.v. with saline supplemented with the anaesthetic. Blood pressure was monitored continuously in the carotid artery. Effects of NPS 2143 administered i.v. as bolus on MAP in the presence and absence of felodypine and losartan were investigated. Both, felodipine and losartan pretreatment provoked a persistent DMAP decrease by 18+/-3 and 14+/-3 mmHg, respectively. Infusion of NPS 2143 at 1 mg/kg b.w. confirmed hypertensive activity of calcilytic and increased blood pressure for 21+/-4 mmHg. In contrast, administration of NPS 2143 in felodipine as well as in losartan pretreated rats did not change DMAP as compared to felodipine/control and losartan/control groups, respectively. Our study indicated that both the blockade of calcium channels and the AT1 receptor activity prevented the hypertensive effect of calcilytic NPS 2143. This finding might be particularly important in understanding the mechanisms that mediated blood pressure changes related to the activity of calcium receptor.
Subject(s)
Blood Pressure/drug effects , Calcium Channels/drug effects , Hypertension/chemically induced , Naphthalenes/pharmacology , Angiotensin II Type 1 Receptor Blockers/pharmacology , Animals , Calcium Channel Blockers/pharmacology , Calcium Channels/metabolism , Felodipine/pharmacology , Losartan/pharmacology , Male , Rats , Rats, Wistar , Receptor, Angiotensin, Type 1/drug effects , Receptor, Angiotensin, Type 1/metabolism , Receptors, Calcium-Sensing/antagonists & inhibitorsSubject(s)
Cardiovascular Diseases/physiopathology , Hypertension/physiopathology , Kidney Diseases/physiopathology , Nephrology , Renin-Angiotensin System/physiology , Angiotensin I/physiology , Angiotensin II Type 1 Receptor Blockers/therapeutic use , Angiotensin-Converting Enzyme Inhibitors/therapeutic use , Animals , Antihypertensive Agents/therapeutic use , Cardiovascular Diseases/drug therapy , Cardiovascular Diseases/prevention & control , Drug Evaluation, Preclinical , Humans , Hypertension/drug therapy , Kidney Diseases/drug therapy , Kidney Diseases/prevention & control , Peptide Fragments/physiology , Randomized Controlled Trials as Topic/statistics & numerical data , Rats , Receptor, Angiotensin, Type 1/drug effects , Receptor, Angiotensin, Type 1/physiology , Receptors, Cell Surface/physiology , Renin/antagonists & inhibitors , Renin/physiology , Renin-Angiotensin System/drug effects , Prorenin ReceptorABSTRACT
Angiotensin II (Ang II) administered intracerebroventriculary (icv) at a dose that induces drinking behavior in rats significantly increased K(+)-stimulated release of [(3)H] noradrenaline (NA) in hypothalamus without affecting basal [(3)H] NA release. The observed difference between the effects of Ang II on basal and K(+)-stimulated [(3)H]NA release may possibly be due to the fact that peptides are released after increased neuronal activity. It can be suggested that Ang II is important primarily in pathological states and that NA plays a substantial role in the brain Ang II-induced drinking response. The imidazolic nonpeptidic compound 2-n-butyl-4-chloro-5-hydroxymethyl-1-{[2-(1H-tetrazol-5-yl)biphenyl-4-yl]methyl}imidazole potassium salt (DuP 753, losartan), its active metabolite 2-n-butyl-4-chloro-1-{[2-(1H-tetrazol-5-yl)- biphenyl-4-yl]methyl}imidazole-5-carboxylic acid (EXP 3174) and peptide Ang II analogue, sarmesin, antagonized the Ang II-induced effect on [(3)H]NA release, in spite of the differences in their chemical structures. Thus, the drugs tested inhibited K(+)-stimulated [(3)H]NA release in hypothalamus, acting via the angiotensin (AT)(1) receptor subtype. We could not reject the possibility of a non-receptor mechanism of action for DuP753, EXP3174 and sarmesin. This research allows us to suggest a neurochemical mechanism for the modulatory role of these drugs on the NA-ergic system. The Ang II receptor antagonists studied may become important therapeutic agents, which act preferentially on pathologically activated systems. These agents may be of use for the prevention of excessive ingestion of water in some neuropsychotic diseases.
Subject(s)
Angiotensin II/pharmacology , Angiotensin Receptor Antagonists , Norepinephrine/metabolism , Receptor, Angiotensin, Type 1/drug effects , Angiotensin II/analogs & derivatives , Angiotensin II/metabolism , Angiotensin II Type 1 Receptor Blockers/pharmacology , Animals , Drinking/drug effects , Hypothalamus/drug effects , Hypothalamus/metabolism , Imidazoles/pharmacology , Injections, Intraventricular , Losartan/pharmacology , Male , Potassium/pharmacology , Rats , Rats, Wistar , Receptor, Angiotensin, Type 1/metabolism , Tetrazoles/pharmacologyABSTRACT
For myocardial regeneration therapy, the low differentiation capability of functional cardiomyocytes sufficient to replace the damaged myocardial tissue is one of the major difficulties. Using Nkx2.5-GFP knock-in ES cells, we show a new efficient method to obtain cardiomyocytes from embryonic stem (ES) cells. The proportion of GFP-positive cells was significantly increased when ES cells were cultured with a conditioned medium from aortic endothelial cells (ECs), accompanied by upregulation of cardiac-specific genes as well as other mesodermal genes. The promotion was more prominent when EC-conditioned medium was added at an early stage of ES cell differentiation culture (Day 0-3). Inhibitors of bone morphogenic protein (BMP), cyclooxygenase (COX), and nitric oxide synthetase (NO) prevented the promotion of cardiomyogenesis by EC-conditioned medium. These results suggest that supplementation of EC-conditioned medium enables cardiomyocytes to be obtained efficiently through promotion of mesoderm induction, which is regulated by BMP, COX, and NOS.
Subject(s)
Cell Culture Techniques , Cell Differentiation , Embryonic Stem Cells/cytology , Endothelium, Vascular/cytology , Endothelium, Vascular/metabolism , Myocytes, Cardiac/cytology , Angiotensin II Type 1 Receptor Blockers/pharmacology , Animals , Aorta/cytology , Bone Morphogenetic Proteins/antagonists & inhibitors , Cell Differentiation/drug effects , Culture Media, Conditioned/metabolism , Culture Media, Conditioned/pharmacology , Cyclooxygenase Inhibitors/pharmacology , Embryonic Stem Cells/drug effects , Endothelin-1/antagonists & inhibitors , Endothelium, Vascular/drug effects , Gene Knock-In Techniques , Green Fluorescent Proteins/genetics , Homeobox Protein Nkx-2.5 , Homeodomain Proteins/genetics , Mice , Myocytes, Cardiac/drug effects , Nitric Oxide Synthase/antagonists & inhibitors , Prostaglandin-Endoperoxide Synthases/drug effects , Receptor, Angiotensin, Type 1/drug effects , Transcription Factors/geneticsABSTRACT
In many traditional schools of medicine it is claimed that a balanced modulation of several targets can provide a superior therapeutic effect and decrease in side effect profile compared to a single action from a single selective ligand, especially in the treatment of certain chronic and complex diseases, such as diabetes and obesity. Diabetes and obesity have a multi-factorial basis involving both genetic and environmental risk factors. A wide array of medicinal plants and their active constituents play a role in the prevention and treatment of diabetes. Salacia roots have been used in Ayurvedic medicine for diabetes and obesity since antiquity, and have been extensively consumed in Japan, the United States and other countries as a food supplement for the prevention of obesity and diabetes. Recent pharmacological studies have demonstrated that Salacia roots modulate multiple targets: peroxisome proliferator-activated receptor-alpha-mediated lipogenic gene transcription, angiotensin II/angiotensin II type 1 receptor, alpha-glucosidase, aldose reductase and pancreatic lipase. These multi-target actions may mainly contribute to Salacia root-induced improvement of type 2 diabetes and obesity-associated hyperglycemia, dyslipidemia and related cardiovascular complications seen in humans and rodents. The results of bioassay-guided identification indicate that mangiferin, salacinol, kotalanol and kotalagenin 16-acetate are at least in part responsible for these multi-target regulatory activities of Salacia roots. The evidence suggests that this unique traditional medicine fulfills a multiple-target strategy in the prevention and treatment of diabetes and obesity. Although toxicological studies have suggested minimal adverse effects of the herbal medicine in rodents, a clinical trial is crucial to further confirm the safety of Salacia roots. In addition, further mechanistic studies are necessary in order to allow a better understanding of how use of Salacia root may interact with other therapeutic interventions.
Subject(s)
Diabetes Mellitus/drug therapy , Medicine, Ayurvedic , Obesity/drug therapy , Phytotherapy , Salacia/chemistry , Aldehyde Reductase/antagonists & inhibitors , Angiotensin II Type 1 Receptor Blockers/pharmacology , Animals , Fatty Acids/metabolism , Glycoside Hydrolase Inhibitors , Humans , Myocardium/metabolism , PPAR alpha/agonists , Pancrelipase/antagonists & inhibitors , Plant Roots/chemistry , Receptor, Angiotensin, Type 1/drug effects , Receptor, Angiotensin, Type 1/physiology , Signal TransductionABSTRACT
Angiotensin II type 1 (AT(1)) receptor antagonists (blockers) [ARBs] are highly selective for the AT(1) receptor and block the diverse effects of angiotensin II. When high BP is not controlled by low-dose ARB monotherapy, physicians need to employ another strategy, either high-dose ARB monotherapy or combination therapy with calcium channel antagonists (blockers) [CCBs], diuretics, or other agents. High-dose ARB monotherapy is more effective for decreasing proteinuria than low-dose ARB monotherapy or CCBs. Although the ARB valsartan has been shown to prevent coronary restenosis in a clinical study (Val-PREST [Valsartan for prevention of restenosis after stenting of type B2/C lesions]), it is still unclear whether ARBs help to prevent restenosis. The results reported by Peters in this issue highlight the relative efficacies of low- (80 mg/day) and high-dose valsartan (160-320 mg/day) for the prevention of in-stent restenosis after the implantation of bare-metal stents, and suggest that high-dose valsartan can reduce the in-stent restenosis rate, target lesion revascularization and target vessel revascularization rates, late lumen loss, and major adverse cardiac events rate more effectively than low-dose valsartan. A better understanding of the differences in the efficacies of high- and low-dose ARBs could be useful in the treatment of patients with cardiovascular disease and may resolve the issue of whether ARBs prevent coronary restenosis. Clinical benefits may be induced by complete blockade of the renin-angiotensin system using high-dose ARB monotherapy. Therefore, physicians need to select a strategy carefully; i.e. either high-dose ARB monotherapy or combination therapy.
Subject(s)
Angiotensin II Type 1 Receptor Blockers/therapeutic use , Cardiovascular Diseases/drug therapy , Angiotensin II Type 1 Receptor Blockers/pharmacology , Cardiovascular Agents/pharmacology , Cardiovascular Agents/therapeutic use , Drug Therapy, Combination , Humans , Receptor, Angiotensin, Type 1/drug effects , Treatment OutcomeABSTRACT
AIMS: We investigated the effect of the water extract of Salacia oblonga (SOE), an ayurvedic antidiabetic and antiobesity medicine, on obesity and diabetes-associated cardiac hypertrophy and discuss the role of modulation of cardiac angiotensin II type 1 receptor (AT(1)) expression in the effect. METHODS: SOE (100 mg/kg) was given orally to male Zucker diabetic fatty (ZDF) rats for 7 weeks. At the end-point of the treatment, the hearts and left ventricles were weighed, cardiomyocyte cross-sectional areas were measured, and cardiac gene profiles were analysed. On the other hand, angiotensin II-stimulated embryonic rat heart-derived H9c2 cells and neonatal rat cardiac fibroblasts were pretreated with SOE and one of its prominent components mangiferin (MA), respectively. Atrial natriuretic peptide (ANP) mRNA expression and protein synthesis and [(3)H]thymidine incorporation were determined. RESULTS: SOE-treated ZDF rats showed less cardiac hypertrophy (decrease in weights of the hearts and left ventricles and reduced cardiomyocyte cross-sectional areas). SOE treatment suppressed cardiac overexpression of ANP, brain natriuretic peptide (BNP) and AT(1) mRNAs and AT(1) protein in ZDF rats. SOE (50-100 microg/ml) and MA (25 micromol) suppressed angiotensin II-induced ANP mRNA overexpression and protein synthesis in H9c2 cells. They also inhibited angiotensin II-stimulated [(3)H]thymidine incorporation by cardiac fibroblasts. CONCLUSIONS: Our findings demonstrate that SOE decreases cardiac hypertrophy in ZDF rats, at least in part by inhibiting cardiac AT(1) overexpression. These studies provide insights into a potential cardioprotective role of a traditional herb, which supports further clinical evaluation in obesity and diabetes-associated cardiac hypertrophy.