RESUMO
The natriuretic peptides are believed to play an important role in the pathophysiology of congestive heart failure (CHF). We utilized a quantitative cytomorphometric method, using double immunocytochemical labeling, to assess the characteristics of atrial natriuretic peptide (ANP) and brain natriuretic peptide (BNP) in atrial granules in an experimental model of rats with CHF induced by aortocaval fistula. Rats with CHF were further divided into decompensated (sodium-retaining) and compensated (sodium-excreting) subgroups and compared with a sham-operated control group. A total of 947 granules in myocytes in the right atrium were analyzed, using electron microscopy and a computerized analysis system. Decompensated CHF was associated with alterations in the modal nature of granule content packing, as depicted by moving bin analysis, and in the granule density of both peptides. In control rats, the mean density of gold particles attached to both peptides was 347.0 +/- 103.6 and 306.3 +/- 89.9 gold particles/microm2 for ANP and BNP, respectively. Similar mean density was revealed in the compensated rats (390.6 +/- 81.0 and 351.3 +/- 62.1 gold particles/microm2 for ANP and BNP, respectively). However, in rats with decompensated CHF, a significant decrease in the mean density of gold particles was observed (141.6 +/- 67.3 and 158.0 +/- 71.2 gold particles/microm2 for ANP and BNP, respectively; p<0.05 compared with compensated rats, for both ANP and BNP). The ANP:BNP ratio did not differ between groups. These findings indicate that the development of decompensated CHF in rats with aortocaval fistula is associated with a marked decrease in the density of both peptides in atrial granules, as well as in alterations in the quantal nature of granule formation. The data further suggest that both peptides, ANP and BNP, may be regulated in the atrium by a common secretory mechanism in CHF.
Assuntos
Fator Natriurético Atrial/metabolismo , Insuficiência Cardíaca/metabolismo , Miocárdio/metabolismo , Peptídeo Natriurético Encefálico/metabolismo , Vesículas Secretórias/metabolismo , Animais , Átrios do Coração/metabolismo , Átrios do Coração/ultraestrutura , Insuficiência Cardíaca/patologia , Processamento de Imagem Assistida por Computador , Imuno-Histoquímica , Masculino , Miocárdio/ultraestrutura , Ratos , Ratos WistarRESUMO
The kidney is both a source of endothelin (ET) generation and an important target organ of this peptide. The highest concentrations of ET-1 in the body exist in the renal medulla, where it mediates natriuretic and diuretic effects through the ET(B) receptor subtype. It is proposed that aberrations in the renal ET system may lead to sodium and water retention and subsequently to the development of hypertension.
Assuntos
Endotelinas/fisiologia , Rim/fisiopatologia , Animais , Diurese/fisiologia , Endotelina-1/fisiologia , Humanos , Hipertensão/fisiopatologia , Medula Renal/fisiopatologia , Natriurese/fisiologia , Receptor de Endotelina B , Receptores de Endotelina/fisiologiaRESUMO
OBJECTIVE: Surgical closure of a large arteriovenous (A-V) fistula in patients and animals is associated with prompt diuresis and natriuresis. However, the mechanisms underlying these changes remained largely unknown. METHODS: The present study evaluated the hormonal balance between major antinatriuretic systems (plasma renin activity, PRA, and arginine vasopressin, AVP) and natriuretic systems (atrial natriuretic peptide, ANP, and renal nitric oxide, NO) in Wistar rats with an A-V fistula (1.2 mm O.D., side to side) between the abdominal aorta and inferior vena cava. RESULTS: The placement of an A-V fistula caused progressive sodium retention (UNaV decreased from 1500 to 100 microequiv./day), a significant drop in mean arterial blood pressure (MAP) from 127+/-3 to 75+/-2 mmHg (P<0.01), and a significant increase in ANP (from 94+/-12 to 389+/-135 pg/ml, P<0.05), PRA (from 22.1+/-2.0 to 47+/-14 ng angiotensin I [Ang I]/ml/h, P<0.05), AVP (from 14.2+/-3.6 to 37.7+/-9.6 pg/ml, P<0.05), norepinephrine (from 184.2+/-40.5 to 1112.6+/-293.2 pg/ml, P<0.05) and epinephrine (from 667.5+/-175.9 to 2049.8+/-496.9 pg/ml, P<0.05). Furthermore, these changes were associated with a 3-fold increase in the renal medullary immunoreactive levels of endothelial NO synthase (eNOS), an endogenous vasodilator that plays an important role in the regulation of medullary blood flow. After 6 days, rats with A-V fistula and maximal sodium retention underwent surgical closure of the A-V fistula. The A-V fistula closure was associated with dramatic natriuresis (UNaV=2563+/-78 and 1918+/-246 microEq/day on days 3 and 6 following the closure, respectively) and restoration of MAP to normal levels (111+/-6 mmHg); PRA decreased to 29+/-5 ng Ang I/ml/h, AVP to 20.3+/-7.1 pg/ml, and medullary eNOS declined to basal levels, whereas plasma ANP concentrations remained elevated (380+/-90 pg/ml) after 3 days and returned to normal (92+/-12 pg/ml) on day 6. CONCLUSIONS: These results demonstrate that the creation of A-V fistula is associated with activation of both natriuretic and antinatriuretic systems. Closure of A-V fistula is characterized by shifting the balance in favor of the natriuretic substances. Moreover, the observed alterations in medullary eNOS following the creation and closure of A-V fistula suggest that this system, an important determinant of medullary blood flow, may contribute significantly to the regulation of sodium excretion in this model.
Assuntos
Insuficiência Cardíaca/fisiopatologia , Natriurese/fisiologia , Animais , Arginina Vasopressina/sangue , Derivação Arteriovenosa Cirúrgica , Fator Natriurético Atrial/sangue , Pressão Sanguínea/fisiologia , Insuficiência Cardíaca/metabolismo , Hormônios/sangue , Rim/enzimologia , Masculino , Óxido Nítrico Sintase/metabolismo , Ratos , Ratos Wistar , Renina/sangue , Sódio/urina , UrinaRESUMO
Endothelin-1 (ET-1) at high concentrations has marked antidiuretic and antinatriuretic activities, whereas its precursor, big endothelin-1 (big ET-1), has surprisingly potent diuretic and natriuretic actions. The mechanisms underlying the excretory effects of big ET-1 have not been fully elucidated. To explore these mechanisms, we examined the effects of a highly selective ET(B) antagonist (A-192621.1), a calcium channel blocker (verapamil), a nitric oxide synthase inhibitor (N-nitro-L-arginine methyl ester [L-NAME]), and a cyclooxygenase inhibitor (indomethacin) on the systemic and renal actions of big ET-1 in anesthetized rats. An intravenous bolus injection of incremental doses of big ET-1 (0.3, 1. 0, and 3.0 nmol/kg) produced a significant hypertensive effect that was dose dependent and prolonged (from 113+/-7 mm Hg to a maximum of 148+/-6 mm Hg). The administration of big ET-1 induced marked diuretic and natriuretic responses (urinary flow rate increased from 8.5+/-1 to 110+/-14 microL/min, and fractional excretion of sodium increased from 0.38+/-0.13% to 7.51+/-1.24%). Glomerular filtration rate and renal plasma flow significantly decreased only at the highest dose of big ET-1. Pretreatment with A-192621.1 (3 mg/kg plus 3 mg. kg(-1). h(-1)) significantly abolished the diuretic (17+/-5 microL/min to a maximum of 19+/-3 microL/min) and natriuretic (0. 29+/-0.1% to a maximum of 1.93+/-0.37%) responses induced by big ET-1. Moreover, A-192621.1 potentiated the decline in glomerular filtration rate and renal plasma flow and the increase in mean arterial blood pressure produced by the low doses of big ET-1. Similar to A-192621.1, pretreatment with a nitric oxide synthase inhibitor (L-NAME, 10 mg/kg plus 5 mg. kg(-1). h(-1)) significantly and comparably reduced the diuretic and natriuretic actions of big ET-1 and augmented the hypoperfusion/hypofiltration and systemic vasoconstriction induced by high doses of the peptide. Pretreatment with verapamil (2 mg. kg(-1). h(-1)) slightly inhibited the diuretic/natriuretic effects of the high-dose big ET-1 and completely prevented the increase in mean arterial blood pressure provoked by the peptide. Unlike verapamil and L-NAME, only indomethacin administration was associated with significant natriuretic/diuretic responses and did not influence the pressor effect and renal actions of big ET-1. Taken together, these results suggest that big ET-1-induced diuretic and natriuretic responses are mediated mainly by stimulation of nitric oxide production coupled to ET(B) receptor subtype activation.
Assuntos
Endotelinas/farmacologia , Natriurese/efeitos dos fármacos , Natriurese/fisiologia , Precursores de Proteínas/farmacologia , Receptores de Endotelina/fisiologia , Animais , Cálcio/metabolismo , Bloqueadores dos Canais de Cálcio/farmacologia , Fármacos Cardiovasculares/farmacologia , Citosol/metabolismo , Antagonistas dos Receptores de Endotelina , Endotelina-1 , Inibidores Enzimáticos/farmacologia , Taxa de Filtração Glomerular/efeitos dos fármacos , Taxa de Filtração Glomerular/fisiologia , Indometacina/farmacologia , Rim/química , Rim/efeitos dos fármacos , Rim/fisiologia , Masculino , NG-Nitroarginina Metil Éster/farmacologia , Óxido Nítrico/metabolismo , Prostaglandinas/metabolismo , Pirrolidinas/farmacologia , Ratos , Ratos Wistar , Receptor de Endotelina B , Micção/efeitos dos fármacos , Micção/fisiologia , Verapamil/farmacologiaRESUMO
Extensive skeletal muscle injury, whether caused by mechanical crush or by extreme physical exertion, is incompatible with life, unless treated early and vigorously. The immediate cause of morbidity is leakiness of the sarcolemmal membrane to cardiotoxic or nephrotoxic cations and metabolites (K, PO4, myoglobin and urate) of the sarcoplasma, and rapid massive uptake by the muscles of extracellular fluid, sodium and calcium, leading to profound hypovolemic and hyocalcemic shock. Casualties who survive the early steep of hyperkalemia and arterial hypotension are susceptible to myoglubinuric acute renal failure owing mainly to the combination of renal vasoconstriction, nephrotoxicity, and tubular obstruction by myoglobin plugs and urate. Management includes immediate (prehospital) intravenous volume replacement followed by mannitol-alkaline diuresis. The alkali regimen ameliorates the acidosis associated with shock and the hyperkalemia, and protects against the nephrotoxicity of myoglobin and urate by alkalinization of the urine. Mannitol, through its impermeant hyperoncotic properties, decompresses and mobilizes muscle edema and promotes renal tubular flow, thus flushing myoglobin plugs and enhancing urinary elimination of nephrotoxic metabolites. With this regimen and when necessary also with the use of dialysis, a substantial salvage of lives, limbs, and kidney function has been achieved recently compared with invariable mortality for casualties who were buried for 3 to 4 hours or more in the early 1940s (World War 2).
Assuntos
Injúria Renal Aguda/etiologia , Síndrome de Esmagamento/complicações , Injúria Renal Aguda/metabolismo , Injúria Renal Aguda/patologia , Trifosfato de Adenosina/metabolismo , Humanos , Óxido Nítrico/fisiologia , Estresse Oxidativo , Rabdomiólise/etiologia , VasoconstriçãoRESUMO
BACKGROUND: Congestive heart failure (CHF) is associated with a decrease in renal perfusion. Because endothelium-derived NO is important in the regulation of renal blood flow (RBF), we tested the hypothesis that an impairment in the NO system may contribute to the decrease in RBF in rats with experimental CHF. METHODS AND RESULTS: Studies were performed in rats with experimental high-output CHF induced by aortocaval (AV) fistula and sham-operated controls. In controls, incremental doses of acetylcholine (ACh, 1 to 100 microg x kg(-1) x min(-1)) increased RBF and caused a dose-related decrease in renal vascular resistance (RVR). However, the increase in RBF and decrease in RVR were markedly attenuated in rats with CHF. Likewise, the effects of ACh on urinary sodium and cGMP excretion were also diminished in CHF rats, as was the renal vasodilatory effect of the NO donor S-nitroso-N-acetylpenicillamine (SNAP). These attenuated responses to endothelium-dependent and -independent renal vasodilators in CHF rats occurred despite a normal baseline and stimulated NO2+NO3 excretion and normal expression of renal endothelial NO synthase (eNOS), as determined by eNOS mRNA levels and immunoreactive protein. Infusion of the NO precursor L-arginine did not affect baseline RBF or the response to ACh in rats with CHF. However, administration of the nonpeptide angiotensin II receptor antagonist A81988 before ACh completely restored the renal vasodilatory response to ACh in CHF rats. CONCLUSIONS: This study demonstrates that despite a significant attenuation in the NO-related renal vasodilatory responses, the integrity of the renal NO system is preserved in rats with chronic AV fistula. This impairment in NO-mediated renal vasodilation in experimental CHF appears to be related to increased activity of the renin-angiotensin system and may contribute further to the decrease in renal perfusion seen in CHF.
Assuntos
Angiotensina II/fisiologia , Fístula Arteriovenosa/fisiopatologia , Insuficiência Cardíaca/fisiopatologia , Óxido Nítrico/fisiologia , Circulação Renal/fisiologia , Vasodilatação/fisiologia , Veias Cavas/fisiopatologia , Acetilcolina/farmacologia , Antagonistas de Receptores de Angiotensina , Animais , Doenças da Aorta/fisiopatologia , Inibidores Enzimáticos/farmacologia , Hemodinâmica/efeitos dos fármacos , Rim/efeitos dos fármacos , Masculino , Óxido Nítrico Sintase/metabolismo , Penicilamina/análogos & derivados , Penicilamina/farmacologia , Ratos , Ratos Wistar , Circulação Renal/efeitos dos fármacos , S-Nitroso-N-Acetilpenicilamina , Vasodilatadores/farmacologiaRESUMO
Increasing evidence suggests that endothelin, a potent vasoconstrictor, is implicated in cyclosporin A (CsA)-induced nephrotoxicity. Increased levels of urinary and circulating endothelin have been described in CsA-treated humans and animals. The exact mechanisms by which CsA induces these increases are still unknown, and no data indicate whether these elevated levels reflect increased synthesis or decreased clearance of endothelin. In the present study, we investigated the effects of CsA administration (50 mg/kg per day i.p. for 6 days) to rats on plasma and urinary levels of endothelin; expression of endothelin-1 (ET-1), ET-3, and endothelin-converting enzyme in renal tissue; clearance of infused 125I-ET-1; and degradation of 125I-ET-1 by recombinant neutral endopeptidase. Rats given CsA for 6 days developed severe renal insufficiency, as shown by a 74% decrease in creatinine clearance rate (Ccr) (P < .006). Ccr was remarkably improved in CsA-treated rats that received bosentan, the combined antagonist of both endothelin A and endothelin B receptors. Urinary excretion of endothelin increased from an undetectable level to 31.7 +/- 6.0 pg/24 h (P < .001), and plasma levels of endothelin were unchanged (2.8 +/- 0.2 to 3.1 +/- 0.2 pg/mL). Reverse transcription followed by quantitative polymerase chain reaction revealed that ET-1 mRNA in the renal medulla increased by 59% (P < .006), whereas the expression of both ET-3 and endothelin-converting enzyme was unchanged. In other rats, neither acute nor chronic treatment with CsA affected either the clearance of 125I-ET-1 from the blood or the renal and pulmonary uptake of the peptide. Moreover, CsA did not affect the degradation of 125I-ET-1 by highly purified recombinant neutral endopeptidase, a well-known endothelinase. Taken together, these data suggest that the elevated urinary endothelin levels obtained after CsA treatment originate from the kidney and reflect increased renal synthesis of ET-1. Moreover, the production of endothelin appears to be regulated at the mRNA transcription level, and expressions of ET-1 and ET-3 are regulated independently.
Assuntos
Ciclosporina/farmacologia , Endotelinas/metabolismo , Rim/metabolismo , Animais , Ácido Aspártico Endopeptidases/genética , Ácido Aspártico Endopeptidases/metabolismo , Sequência de Bases , Bosentana , Creatinina/metabolismo , Enzimas Conversoras de Endotelina , Endotelinas/genética , Endotelinas/urina , Rim/efeitos dos fármacos , Masculino , Metaloendopeptidases/genética , Metaloendopeptidases/metabolismo , Sondas Moleculares/genética , Dados de Sequência Molecular , Neprilisina/metabolismo , Ratos , Ratos Sprague-Dawley , Proteínas Recombinantes , Sulfonamidas/farmacologiaAssuntos
Antimetabólitos Antineoplásicos/efeitos adversos , Leucemia Linfocítica Crônica de Células B/tratamento farmacológico , Síndrome de Lise Tumoral/etiologia , Fosfato de Vidarabina/análogos & derivados , Injúria Renal Aguda/etiologia , Injúria Renal Aguda/prevenção & controle , Alopurinol/uso terapêutico , Antimetabólitos Antineoplásicos/uso terapêutico , Terapia Combinada , Feminino , Hidratação , Humanos , Leucemia Linfocítica Crônica de Células B/patologia , Leucopenia/etiologia , Pessoa de Meia-Idade , Bicarbonato de Sódio/uso terapêutico , Síndrome de Lise Tumoral/prevenção & controle , Fosfato de Vidarabina/efeitos adversos , Fosfato de Vidarabina/uso terapêuticoRESUMO
BACKGROUND: Chronic activation of the renin-angiotensin system (RAS) plays an important role in the pathogenesis of heart failure. Increasing evidence indicates that other than the circulating RAS, a local RAS exists in several tissues, including the heart. The present study was carried out to quantify cardiac, renal, and pulmonary mRNA levels of renin, angiotensin-converting enzyme (ACE), and types 1 and 2 angiotensin II receptors (AT-1 and AT-2), in rats with different severities of heart failure. METHODS AND RESULTS: Heart failure was induced by the creation of an aortocaval fistula below the renal arteries. Rats with aortocaval fistula either compensate and maintain a normal sodium balance or decompensate and develop severe sodium retention. Six days after placement of the aortocaval fistula, heart weight (normalized to body weight) increased 35% (P < .05) in compensated and 65% in decompensated rats compared with control rats. Plasma renin activity increased 45% (P < .05) in rats in sodium balance and 127% in sodium-retaining rats. Total RNA was extracted from the heart, kidneys, and lungs, followed by reverse transcription-quantitative polymerase chain reaction. Renin mRNA levels in the heart, after 40 cycles, increased 68% (P < .01) and 140% in rats with either compensated or decompensated heart failure, respectively. Renal renin-mRNA levels also increased 130% (P < .05) in decompensated and only 52% (P < .05) in compensated animals. ACE-mRNA increased in a similar pattern in the heart but not in either the kidneys or lungs. Moreover, pulmonary, renal, and cardiac ACE immunoreactivity levels, assessed by Western blot analysis, showed the same trend. AT-1 receptor mRNA levels decreased 54% (P < .05) only in the myocardium of decompensated rats, whereas AT-2 receptor mRNA did not change in any tissue studied. CONCLUSIONS: The development of heart failure is associated with a remarkable increase in the expression of a local RAS in the heart, which may contribute to the pathogenesis of this clinical syndrome.
Assuntos
Insuficiência Cardíaca/metabolismo , Rim/metabolismo , Pulmão/metabolismo , Miocárdio/metabolismo , Sistema Renina-Angiotensina/fisiologia , Angiotensina II/biossíntese , Animais , Western Blotting , Expressão Gênica , Insuficiência Cardíaca/etiologia , Masculino , Peptidil Dipeptidase A/biossíntese , Reação em Cadeia da Polimerase/métodos , RNA Mensageiro/genética , Ratos , Ratos Sprague-Dawley , Receptores de Angiotensina/biossíntese , Receptores de Angiotensina/classificação , Renina/biossíntese , Sistema Renina-Angiotensina/genéticaRESUMO
Congestive heart failure is characterized by avid sodium retention and a blunted renal response to exogenous and endogenous atrial natriuretic peptide. Inhibition of neutral endopeptidase EC 3.4.24.11, the main enzyme that degrades natriuretic peptides, produces a natriuretic response in different models of congestive heart failure. This raises the possibility that an increase in either the expression or activity of neutral endopeptidase is responsible for these phenomena. In the present study, we examined (1) the renal effects of SQ-28,603, a neutral endopeptidase inhibitor, in rats with moderate and severe congestive heart failure induced by an aortocaval fistula compared with sham controls, and (2) neutral endopeptidase expression and activity in the lungs and kidneys of these rats. Infusion of SQ-28,603 (40 mg/kg IV) induced a significant natriuretic response in normal rats and rats with moderate congestive heart failure. This response was blunted in rats with severe congestive heart failure. Surprisingly, renal neutral endopeptidase mRNA levels, assessed by quantitative reverse transcriptase-polymerase chain reaction; protein levels, assessed by Western blotting; and activity, assessed by gelatin gels, were comparable in all groups. Pulmonary neutral endopeptidase mRNA levels decreased by 45% in rats with severe congestive heart failure but not in rats with mild congestive heart failure. In addition, pulmonary neutral endopeptidase immunoreactivity levels and activity were significantly decreased in congestive heart failure in correlation with the severity of the disorder.(ABSTRACT TRUNCATED AT 250 WORDS)
Assuntos
Insuficiência Cardíaca/enzimologia , Rim/enzimologia , Pulmão/enzimologia , Neprilisina/fisiologia , Alanina/análogos & derivados , Alanina/farmacologia , Animais , Sequência de Bases , Rim/efeitos dos fármacos , Masculino , Dados de Sequência Molecular , Neprilisina/antagonistas & inibidores , Neprilisina/genética , Reação em Cadeia da Polimerase , RNA Mensageiro/análise , Ratos , Ratos Sprague-DawleyAssuntos
Endotelinas/deficiência , Hipertensão/metabolismo , Rim/metabolismo , Animais , Humanos , CamundongosRESUMO
The role of angiotensin II (ANG II) in the development of isoproterenol (Iso)-induced cardiac hypertrophy was examined in rats. Iso increased cardiac mass, left ventricular RNA-to-DNA ratio, and the cardiac content of both myosin heavy chain and hydroxyproline in a dose-dependent manner, indicating that Iso-induced cardiac hypertrophy involves growth of both muscle and connective tissue. Cardiac hypertrophy reverted within 11-14 days after cessation of Iso. Propranolol prevented development of Iso-induced cardiac hypertrophy but did not affect the rate of its reversal. The ANG II receptor blocker losartan (Los) did not significantly decrease the hypertrophic response to Iso. Los injected after cessation of Iso dramatically enhanced the reversal of cardiac hypertrophy, even in rats that received Los with Iso during the induction of Iso-induced cardiac hypertrophy. ANG II, injected continuously at a subpressor dose that did not affect heart weight when given alone, inhibited reversal of cardiac hypertrophy when given after cessation of Iso. Los did not significantly affect the induction of the protooncogene c-fos by Iso. We conclude that endogenous ANG II has a major function in maintaining Iso-induced cardiac hypertrophy but does not mediate its induction. This suggests that different interactive stimuli may be required for development of cardiac hypertrophy, i.e., for initiation and for maintenance.
Assuntos
Angiotensina II/farmacologia , Compostos de Bifenilo/farmacologia , Cardiomegalia/fisiopatologia , Imidazóis/farmacologia , Isoproterenol/farmacologia , Tetrazóis/farmacologia , Actinas/biossíntese , Angiotensina II/antagonistas & inibidores , Animais , Sequência de Bases , Pressão Sanguínea/efeitos dos fármacos , Captopril/farmacologia , Cardiomegalia/induzido quimicamente , Cardiomegalia/tratamento farmacológico , DNA/metabolismo , Primers do DNA , Relação Dose-Resposta a Droga , Expressão Gênica/efeitos dos fármacos , Genes fos , Coração/efeitos dos fármacos , Coração/fisiopatologia , Hidralazina/farmacologia , Hidroxiprolina/metabolismo , Labetalol/farmacologia , Losartan , Masculino , Metildopa/farmacologia , Dados de Sequência Molecular , Miocárdio/metabolismo , Miosinas/biossíntese , Reação em Cadeia da Polimerase , RNA/metabolismo , RNA Mensageiro/biossíntese , Ratos , Ratos Sprague-Dawley , Transcrição Gênica/efeitos dos fármacosRESUMO
1. Urodilatin is a 32 amino-acid peptide of similar sequence to atrial natriuretic peptide (ANP), with four additional amino-acids at the N-terminus. Although ANP and urodilatin bind to the same receptors with similar affinities, urodilatin is more active than ANP as a natriuretic agent. Previous studies, using neutral endopeptidase EC 3.4.24.11 (NEP) derived from crude membrane preparations, were inconclusive, but suggested that urodilatin was more resistant than ANP to degradation by this enzyme. In the present study, we compared the degradation rates of [125I]-urodilatin and [125I]-ANP by pure recombinant NEP (rNEP). 2. Incubation of radioactively labelled ANP with rNEP resulted in a much more rapid degradation of the peptide than that for labelled urodilatin. 3. Both phosphoramidon and SQ-28,603, potent inhibitors of NEP, completely protected both peptides from metabolism by rNEP. 4. The circular dichroism spectra of the two peptides indicate that they are very similar and exist largely in unordered or flexible conformations. 5. These results support the relative resistance of urodilatin to NEP, and indicate that urodilatin may be of use as a therapeutic agent, in conditions in which ANP is ineffective.
Assuntos
Fator Natriurético Atrial/metabolismo , Diuréticos/metabolismo , Neprilisina/metabolismo , Fragmentos de Peptídeos/metabolismo , Dicroísmo Circular , Humanos , Hidrólise , Radioisótopos do Iodo , Cinética , Neprilisina/antagonistas & inibidores , Proteínas Recombinantes/metabolismo , Ácido Tricloroacético/químicaRESUMO
During severe congestive heart failure (CHF), a number of sodium-retaining and vasoconstricting mechanisms are activated, including the renin-angiotensin-aldosterone system. In CHF, the renal effects of atrial natriuretic factor (ANF) are attenuated. The interaction of these endocrine factors is a major determinant of the clinical course of CHF. This study was designed to evaluate the role of the renin-angiotensin-aldosterone system in the development of avid sodium retention in CHF, induced in rats by creation of an aorto-caval fistula. Rats with aorto-caval fistula either compensate and maintain a normal sodium balance (UNaV > 1400 microEq/day) or decompensate and develop severe sodium retention (UNaV < 200 microEq/day), which leads to severe CHF. Chronic treatment with losartan, an angiotensin II receptor blocker, 10 mg/day, resulted in dramatic natriuresis (UNaV > 1000 microEQ/day) in decompensated rats, but not in compensated rats or controls. ANF infusion (50 micrograms/kg/hr) increased fractional sodium excretion 46-fold in compensated rats, but only 18-fold in decompensated rats. A similar pattern of responsiveness to ANF was observed in urinary cyclic GMP excretion. Chronic losartan treatment restored the natriuretic and urinary cyclic GMP excretion responses of decompensated rats to ANF. The improvement in the natriuretic response after losartan treatment was associated with a suppression of the previously elevated plasma aldosterone. These results demonstrate the pivotal role of angiotensin II in the development of sodium retention and of the blunted renal response to ANF in CHF, and indicate why losartan is useful therapy for cardiac edema.
Assuntos
Antagonistas de Receptores de Angiotensina , Fator Natriurético Atrial/farmacologia , Compostos de Bifenilo/farmacologia , Insuficiência Cardíaca/fisiopatologia , Imidazóis/farmacologia , Natriurese/efeitos dos fármacos , Tetrazóis/farmacologia , Animais , Fator Natriurético Atrial/fisiologia , GMP Cíclico/metabolismo , Interações Medicamentosas , Insuficiência Cardíaca/metabolismo , Rim/efeitos dos fármacos , Losartan , Masculino , Ratos , Ratos Wistar , Sistema Renina-Angiotensina/efeitos dos fármacos , Sistema Renina-Angiotensina/fisiologia , Sódio/metabolismoRESUMO
Endothelin (ET) is a powerful vasoconstrictor peptide synthesized and secreted by the vascular endothelium. Significant amounts of ET are also produced by nonendothelial cells, mainly tubular-epithelial and mesangial cells. Large amounts of ET are found in the urine compared with the small amounts present in blood. Because most of the ET filtered from plasma is subject to degradation by neutral endopeptidase (EC 3.4.24.11) in the proximal tubule, urinary ET is probably of renal origin. The range of urinary ET excretion in healthy persons is 20 to 90 ng/day. The excretion of endothelin is modulated by several mechanical and chemical stimuli such as angiotensin II, phenylephrine, radiocontrast media, cyclosporine, and cis-platin. In addition, enhanced urinary ET excretion has been found in several forms of renal failure, both acute and chronic, and in diabetes mellitus. Thus, urinary ET has the potential of serving as a marker for renal disease.
Assuntos
Endotelinas/urina , Nefropatias/urina , Sequência de Aminoácidos , Animais , Biomarcadores/urina , Humanos , Dados de Sequência MolecularRESUMO
Renal synthesis of a peptide homologous to atrial natriuretic factor (ANF) has been demonstrated. The aim of the present study was to determine if transcription of the ANF gene occurs in the kidney. Rat renal RNA was extracted from whole kidneys, and, separately, from the cortex and outer and inner medulla of rat kidneys. Probing with rat ANF-cDNA did not reveal a detectable message in Northern blot analysis, even when large quantities of RNA were used at low stringency hybridization conditions. Therefore, reverse transcription (RT) followed by 35 cycles of polymerase chain reaction (PCR) was used to search for the renal message for ANF. Two 21-mer primers encompassing the 450 base pairs (bp) of the coding region of the gene were used. Each cycle consisted of annealing at 56 degrees C, extension at 72 degrees C, and denaturation at 94 degrees C. The PCR product was proven to be identical to the ANF gene by high stringency hybridization, which revealed the expected 450-bp hybrid band. Furthermore, the sequence of this product was identical to that of the coding region of the ANF gene. We used an RNA-specific PCR to obtain this band as a single reaction product. We conclude that the transcript of the ANF gene exists in the kidney, at extremely low levels. The low abundance of the RNA message raises major concerns about its physiologic relevance. Direct evidence for the translation of this transcript, and its quantification and localization, is still required to determine its significance.
Assuntos
Fator Natriurético Atrial/genética , Rim/metabolismo , Transcrição Gênica , Animais , Sequência de Bases , Northern Blotting , Feminino , Masculino , Dados de Sequência Molecular , Reação em Cadeia da Polimerase , Ratos , Ratos Sprague-DawleyRESUMO
1. Inhibitors of neutral endopeptidase EC.3.4.24.11 (NEP) have been shown to attenuate the hypertensive effect of big-endothelin-1 (BET-1) in rats. To determine whether NEP converts BET-1 to endothelin-1 (ET-1), the effect of a recombinant NEP (rNEP) on BET-1 and on ET-1 was assessed in vitro. 2. Incubation of [125I]-ET-1 with 1 microgram ml-1 of rNEP resulted in degradation of the peptide within minutes. Increase in the amount of rNEP to 10 micrograms ml-1 led to total cleavage of [125I]-ET-1 within seconds. 3. Phosphoramidon (10 microM) or SQ-28,603 (100 microM) totally suppressed the degradation of [125I]-ET-1 by rNEP. 4. The degradation of [125I]-BET-1 by either 1 or 10 micrograms ml-1 of rNEP was much slower than that of [125I]-ET-1. Again, both phosphoramidon and SQ 28,603 protected the peptide from degradation. 5. Intact [125I]-ET-1 was not observed when [125I]-BET-1 was incubated with rNEP. 6. These data show that neutral endopeptidase EC.3.4.24.11 is not an endothelin converting enzyme.
Assuntos
Alanina/análogos & derivados , Endotelinas/metabolismo , Neprilisina/metabolismo , Precursores de Proteínas/metabolismo , Cromatografia Líquida de Alta Pressão , DNA Complementar/metabolismo , Endotelina-1 , Glicopeptídeos/farmacologia , Humanos , Técnicas In Vitro , Radioisótopos do Iodo , Neprilisina/antagonistas & inibidores , Neprilisina/farmacologia , Proteínas Recombinantes/metabolismoRESUMO
Rats with aortocaval fistula, an experimental model of congestive heart failure (CHF), display two distinct patterns of sodium excretion: some rats develop marked sodium retention and worsening edema with urinary excretion of sodium (UNaV) < 200 microEq per 24 hours, i.e., uncompensated CHF, whereas in others sodium balance rapidly returns to normal (UNaV > 1,400 microEq per 24 hours), i.e., compensated CHF. Similar patterns of sodium excretion are found in patients with CHF. The mechanisms underlying these responses are not fully understood. The present study was designed to assess whether bradykinin plays a role in the compensatory response to CHF. Infusions of either 10 or 50 micrograms/kg per minute of synthetic atrial natriuretic factor (ANF)8-33 into sham-operated control animals produced significant increases in urine flow and fractional excretion of sodium (FENa). Infusions of ANF at the same doses into rats with compensated CHF increased FENa from 0.11 +/- 0.03% to a maximum of 6.10 +/- 1.30%, whereas the rise in FENa in animals with uncompensated CHF was significantly reduced (0.05 +/- 0.01% to 0.59 +/- 0.18%) compared with sham-operated controls (0.23 +/- 0.05% to 8.32 +/- 1.0%) or the group with compensated CHF. Treatment of the compensated rats with the bradykinin antagonist HOE-140 (D-Arg,[Hyp3, Thi5, D-Tic7, Oic8]-bradykinin) given at a rate of 100 nmol/kg per hour did not affect their renal response to the ANF. In addition, infusion of the bradykinin antagonist alone into compensated rats with aortocaval fistula had no significant effect on their basal urinary flow rate or sodium excretion during the infusion.(ABSTRACT TRUNCATED AT 250 WORDS)
Assuntos
Fator Natriurético Atrial/farmacologia , Bradicinina/fisiologia , Insuficiência Cardíaca/fisiopatologia , Natriurese/fisiologia , Adaptação Fisiológica , Animais , Aorta/cirurgia , Derivação Arteriovenosa Cirúrgica , Bradicinina/análogos & derivados , Bradicinina/antagonistas & inibidores , Bradicinina/farmacologia , Diurese/efeitos dos fármacos , Relação Dose-Resposta a Droga , Masculino , Potássio/urina , Ratos , Ratos Endogâmicos , Sódio/metabolismo , Veias Cavas/cirurgiaRESUMO
The regulation of the urinary excretion of endothelin (UETV) and its clinical significance has not yet been established. The present study was designed to examine the effect of angiotensin II (A-II), arginine vasopressin (AVP), and nifedipine on UETV. Anesthetized Munich-Wistar rats were infused with low (50 ng/kg/min) and high (500 ng/kg/min) doses of A-II for 30 min. Both doses significantly increased UETV, from nondetectable (ND) levels to 155 +/- 54 (P < .03) and 450 +/- 86 fg/min (P < .001), respectively. This effect was accompanied by a significant increase in urine flow (UV), from 6 +/- 1 to 67 +/- 12 and 89 +/- 10 microL/min, and in mean arterial pressure (MAP), from 139 +/- 4 to 187 +/- 5 and 217 +/- 3 mm Hg. Infusions of A-II with its nonspecific antagonist, saralasin, resulted in a further increase in UETV to 647 +/- 126 and 782 +/- 117 fg/min (P < .002), respectively. However, infusion of A-II with its specific antagonist, losartan, completely blocked its stimulatory effect on UETV. Infusion of AVP, 10 or 100 mU/kg/h, produced increases in MAP, from 134 +/- 3 to 165 +/- 7 and 203 +/- 4 mm Hg, and in UV from 6 +/- 1 to 37 +/- 6 and 97 +/- 17 microL/min, comparable to A-II, but AVP did not have a marked effect on UETV.(ABSTRACT TRUNCATED AT 250 WORDS)
Assuntos
Angiotensina II/farmacologia , Arginina Vasopressina/farmacologia , Endotelinas/urina , Nifedipino/farmacologia , Análise de Variância , Animais , Anti-Hipertensivos/farmacologia , Compostos de Bifenilo/farmacologia , Pressão Sanguínea/efeitos dos fármacos , Pressão Sanguínea/fisiologia , Captopril/farmacologia , Relação Dose-Resposta a Droga , Taxa de Filtração Glomerular/efeitos dos fármacos , Taxa de Filtração Glomerular/fisiologia , Imidazóis/farmacologia , Rim/metabolismo , Rim/fisiologia , Losartan , Masculino , Radioimunoensaio , Ratos , Ratos Wistar , Saralasina/farmacologia , Tetrazóis/farmacologiaRESUMO
Urodilatin is a new member of the family of natriuretic peptides. It is of renal origin. Previous reports indicate that urodilatin is natriuretic in lower doses than atrial natriuretic factor (ANF)-(99-126) and that it might be more effective than ANF in the treatment of cardiovascular edema. The present study was designed to compare the pharmacokinetics of the hydrolysis and clearance of 125I-labeled urodilatin and 125I-ANF. In control rats, the volume of distribution (Vss), metabolic clearance rate (MCR), and distribution half-life (distribution t1/2) of urodilatin in plasma were not significantly different from those of ANF. Infusion of clearance (c)ANF-(4-23), a specific ligand for receptors that clear ANF in excess amounts (i.e., a bolus injection of 100 micrograms/kg followed by a continuous infusion of 10 micrograms.kg-1 x min-1), increased the amount of intact peptide in the plasma to the same extent for both urodilatin and ANF. In addition, cANF-(4-23) decreased the Vss and the MCR and increased the distribution t1/2 of both peptides to about the same degree. Prior treatment of rats with SQ-28,603, a specific neutral endopeptidase (NEP; EN 3.4.24.11) inhibitor, was without significant effect on the metabolic clearance of urodilatin, whereas it decreased the clearance of ANF by 65%. Furthermore, an infusion of SQ-28,603 suppressed the appearance of the hydrolytic products of ANF in blood but not of urodilatin. Moreover, the inhibitor increased the total amount of ANF recovered in the kidneys to five times the control values, whereas it did not alter the renal uptake of urodilatin.(ABSTRACT TRUNCATED AT 250 WORDS)