RESUMO
Previous studies have provided suggestive evidence for an interaction between ras activation and signalling pathways involved in agonist-stimulated arachidonic acid release in a variety of cell systems. In order to clarify this interaction, we have measured epidermal growth factor (EGF)-stimulated arachidonic acid release in rat-1 fibroblasts transfected with the N-17 dominant negative mutation of ras. Cells transfected with the N-17 ras mutant, display a markedly attenuated arachidonic acid-release response to EGF, compared to sham-transfected and non-transfected cells. In contrast, the response to phorbol myristate acetate (PMA) was not attenuated in the N-17-mutant expressing cells. No differences were detected between sham-transfected and N-17 mutant expressing cells in levels of immunodetectable EGF receptor, cytosolic phospholipase A2 or mitogen-activated protein (MAP) kinase. Attenuation of EGF-stimulated arachidonic acid release in the N-17 mutant expressing cells, was accompanied by a marked diminution in EGF-stimulated tyrosine phosphorylation of MAP kinase. We conclude that the signalling pathway involved in epidermal growth factor-stimulated arachidonic acid release is similar to the signalling pathway for mitogenic responses to epidermal growth factor and requires ras activation, likely followed by a downstream cascade of kinases eventuating in MAP kinase activation.
Assuntos
Ácidos Araquidônicos/metabolismo , Fator de Crescimento Epidérmico/farmacologia , Fibroblastos/citologia , Fibroblastos/metabolismo , Genes ras/fisiologia , Animais , Proteínas Quinases Dependentes de Cálcio-Calmodulina/análise , Proteínas Quinases Dependentes de Cálcio-Calmodulina/metabolismo , Proteínas Quinases Dependentes de Cálcio-Calmodulina/fisiologia , Linhagem Celular , Receptores ErbB/análise , Fibroblastos/química , Regulação da Expressão Gênica/genética , Genes ras/genética , Mutação/genética , Fosfolipases A/análise , Fosfolipases A2 , Fosforilação , Testes de Precipitina , Ratos , Acetato de Tetradecanoilforbol/farmacologia , Transfecção , Trítio , Tirosina/metabolismoRESUMO
Resistance to the natriuretic action of atrial natriuretic factor (ANF) in cirrhosis with ascites has been correlated with rising levels of antinatriuretic factors, such as renin, angiotensin II (AII), and aldosterone, as well as increased sympathetic nerve activity. To determine whether AII can serve as a mediator rather than only as a marker of the antinatriuresis, a nonpressor dose of AII (5 ng/kg/min) was given during an ANF infusion in eight patients with cirrhosis and ascites who responded to ANF infusion with a natriuresis. Patients were maintained in metabolic balance and measurements of para-aminohippuric acid, inulin, and lithium clearance were taken before and during infusion of ANF with or without AII. Atrial natriuretic factor infusion was associated with a natriuretic response accompanied by an increase in glomerular filtration rate, filtration fraction, and lithium clearance compared with baseline. The addition of AII was associated with a return of the glomerular filtration rate to baseline, with no change in filtration fraction. This was reversible on withdrawal of AII infusion. Natriuresis induced by ANF occurred despite baseline elevations of the renin angiotensin aldosterone system and was associated with an increase in distal delivery of sodium and a decrease in fractional reabsorption of distally delivered sodium as estimated by lithium clearance parameters. Angiotensin II infusion exerted effects on both proximal and distal nephron sites to abrogate ANF-induced natriuresis. These results suggest that AII may serve as a mediator as well as a marker of resistance to the natriuretic effect of ANF in patients with cirrhosis and ascites.(ABSTRACT TRUNCATED AT 250 WORDS)
Assuntos
Angiotensina II/fisiologia , Fator Natriurético Atrial/fisiologia , Cirrose Hepática Alcoólica/fisiopatologia , Natriurese/fisiologia , Adulto , Idoso , Ascite/fisiopatologia , Humanos , Masculino , Pessoa de Meia-IdadeRESUMO
The nature of sodium retention in cirrhosis complicated by ascites has been studied for the last 30 years. Resistance to the natriuretic action of atrial natriuretic peptide (ANP) may play a potential role in this sodium retention. To further evaluate this possibility, we studied 12 patients with biopsy-proven cirrhosis and ascites on 2 consecutive days after a 7-day period off diuretics while receiving a 20 mmol/day sodium restricted diet. Following a crossover design, patients underwent head-out water immersion (HWI) for 3 h and were infused with a alpha-human ANP for 2 h on 2 consecutive days. Blood and urine samples were collected hourly. Five patients displayed a natriuretic response to HWI, sufficient to achieve negative sodium balance, and these patients were termed responders. Each of these five patients also displayed a natriuretic response to ANP infusion. In contrast, the other seven patients (nonresponders) consistently failed to develop a natriuretic response to either maneuver. The two groups had similar elevations in plasma ANP concentrations, but at baseline differed in terms of plasma sodium, plasma renin activity, and serum aldosterone. Despite higher serum aldosterone concentrations, nonresponders excreted less potassium than responders during the peak effect of the interventions, suggesting greater sodium delivery to the aldosterone-sensitive nephron segment in responders. We conclude that the inability to mount an adequate sodium excretory response to HWI in patients with cirrhosis may be conveyed through increased antinatriuretic factors that decrease the sodium delivery to the medullary collecting duct and inhibit the natriuretic effect of ANP at that site.
Assuntos
Fator Natriurético Atrial/farmacologia , Imersão , Cirrose Hepática/fisiopatologia , Cirrose Hepática/terapia , Natriurese/efeitos dos fármacos , Aldosterona/sangue , Fator Natriurético Atrial/sangue , Resistência a Medicamentos , Feminino , Humanos , Imersão/fisiopatologia , Infusões Intravenosas , Cirrose Hepática/urina , Masculino , Natriurese/fisiologia , Néfrons/fisiopatologia , Potássio/urina , Renina/sangue , Sódio/sangue , Sódio/urina , Água/metabolismoRESUMO
Our purpose was to quantitate the proportion of H+ removed by the bicarbonate buffer system (BBS) when a modest load of H+ was infused acutely. In addition, the quantitative impact of hyperventilation on the BBS in the extracellular fluid (ECF) and other compartments in this setting was assessed. Three groups of rats (399 +/- 3 g) were anesthetized and connected to a respirator to control their arterial PCO2 and to collect expired air. Metabolic acidosis (pH 7.26 +/- 0.01, bicarbonate 18 +/- 1 mM) was induced by infusion of HCl (0.15 M, 4 mmol/kg) over 60 min, and expired air was collected for two 20-min periods beginning 75 and 105 min after the start of the infusion of HCl in each group. Each rat served as its own control for the rate of production of CO2 from metabolism. The first two groups were time controls. Their arterial PCO2 was constant at either ambient (50 mmHg) or hyperventilation levels (30 mmHg) during both collections (n = 5 each). In the experimental group (n = 5), the PCO2 was decreased from 40 to 27 mmHg during the second collection. The rate of production of CO2 from metabolism did not rise in the second collection in the time control experiments (change = -13.4 +/- 1.7 and -1.4 +/- 2.5 mumol/min, respectively), whereas more CO2 was collected during the second period in the experimental group (change = 42 +/- 9 mumol/min, P = 0.02).(ABSTRACT TRUNCATED AT 250 WORDS)
Assuntos
Bicarbonatos/metabolismo , Ácido Clorídrico/farmacologia , Hiperventilação/metabolismo , Membranas Intracelulares/metabolismo , Equilíbrio Ácido-Base , Acidose/induzido quimicamente , Acidose/metabolismo , Animais , Soluções Tampão , Eletrólitos/sangue , Espaço Extracelular/metabolismo , Masculino , Ratos , Ratos EndogâmicosRESUMO
1. The effect of sodium intake on the natriuresis and hormonal changes induced by head-out water immersion was studied in seven normal subjects during head-out water immersion and on a control day while successively on 20 mmol of sodium/day and 100 mmol of sodium/day diets. The effects of head-out water immersion were compared with those seen on the control day for both diets. 2. The natriuresis on the 100 mmol of sodium/day diet was significantly greater than on the 20 mmol of sodium/day diet (natriuretic peak: 10.3 +/- 2.2 versus 3.9 +/- 1 mmol of sodium/h; P less than 0.01). The total sodium excretion during the 3 h of head-out water immersion was 26.2 +/- 2.0 mmol on the 100 mmol of sodium/day diet and 9.9 +/- 0.9 mmol on the 20 mmol of sodium/day diet (P less than 0.01). In contrast, the increase in the plasma atrial natriuretic factor level was similar on both diets (peak plasma atrial natriuretic factor level 23.1 +/- 1.9 versus 26.2 +/- 1 pg/ml; not significant). As expected, the baseline serum aldosterone level was higher on the 20 mmol of sodium/day diet and, despite a significant suppression, remained significantly higher at the end of the third hour of head-out water immersion (peak serum aldosterone level: 495 +/- 130 versus 197 +/- 26 pmol/l, P less than 0.06). Furthermore, there was an inverse relationship between the serum aldosterone level and the urinary sodium excretion at the time of peak natriuresis (r = -0.59, P less than 0.01). 3. We conclude that the effect of sodium intake on the natriuresis induced by head-out water immersion is more dependent upon anti-natriuretic agents, such as aldosterone, than on natriuretic factors, such as atrial natriuretic factor.
Assuntos
Aldosterona/sangue , Fator Natriurético Atrial/sangue , Imersão/fisiopatologia , Natriurese/fisiologia , Sódio na Dieta/administração & dosagem , Adulto , Humanos , MasculinoRESUMO
Despite intensive investigation, the pathogenesis of sodium retention in patients with chronic liver disease is not fully known. We have studied 19 chronic liver disease patients, 13 without (group 1) and six with (group 2) histories of clinical sodium retention (ascites or edema) by varying dietary sodium intake. The patients were placed on a 20 mmol/day constant diet for 1 wk, followed by a constant 100 mmol/day sodium diet for 1 wk under strict metabolic conditions. After 5 days of equilibration on each diet, blood and urine samples were collected for plasma atrial natriuretic factor levels and urinary sodium excretion. Group 1 patients (n = 6) achieved near sodium balance in 5 days on both a 20-mmol (urinary sodium output = 17 +/- 3 mmol/day) and a 100-mmol sodium diet (urinary sodium output = 80 +/- 5 mmol/day). Atrial natriuretic factor levels in these patients tended to be elevated, but the increase was not significantly greater than that in normal control subjects (10 +/- 4 pg/ml to 19 +/- 4 pg/ml) on the same diets. In contrast, group 2 patients (n = 5) were in significant positive sodium balance on both the 20 mmol/day sodium diet (mean urinary sodium output = 9.5 +/- 3.3 mol/day) and the 100 mmol/day sodium diet (urinary sodium output = 37 +/- 13 mmol/day). This occurred despite significantly elevated baseline atrial natriuretic factor levels and a significant increase in plasma atrial natriuretic factor levels after sodium challenge (62 +/- 9 pg/ml, p less than 0.05) on a 100 mmol/day sodium diet.(ABSTRACT TRUNCATED AT 250 WORDS)
Assuntos
Fator Natriurético Atrial/sangue , Cirrose Hepática/metabolismo , Sódio na Dieta/administração & dosagem , Sódio/urina , Ascite/etiologia , Ascite/metabolismo , Pressão Sanguínea , Doença Crônica , Edema/etiologia , Edema/metabolismo , Átrios do Coração/fisiopatologia , Humanos , Hipertensão Portal/complicações , Hipertensão Portal/fisiopatologia , Rim/metabolismo , Cirrose Hepática/complicações , Fatores de TempoRESUMO
PURPOSE: It is possible that abnormalities in atrial natriuretic peptide may be involved in the pathogenesis of sodium retention in edema states. We performed a study in a group of 12 sodium-retaining cirrhotic subjects to determine the role of this peptide in mediating differences in the natriuretic response to central volume expansion induced by head-out water immersion. PATIENTS AND METHODS: Each patient was maintained for seven days on a 20-mmol sodium intake, and then studied on both control and immersion days. On each day, measurements of the following were obtained: plasma atrial natriuretic peptide, hematocrit, electrolytes, creatinine, plasma renin activity, serum aldosterone, urinary cyclic guanosine monophosphate (cGMP), blood pressure, and pulse rate. RESULTS: In six subjects, immersion resulted in a marked natriuresis sufficient to induce negative sodium balance by the third hour, and these subjects were termed "responders." In these six patients, baseline pre-immersion levels of plasma renin activity and serum aldosterone were all below 3 ng/liter/second and 4 nmol/liter, respectively. In the other six subjects, the natriuretic response to immersion was markedly blunted and insufficient to induce negative sodium balance, and these subjects were termed "non-responders." In these subjects, baseline pre-immersion levels of plasma renin activity and aldosterone were all above 3.5 ng/liter/second and 5 nmol/liter, respectively, and were significantly elevated compared with the responders, and compared with the normal range for control subjects consuming the same sodium intake. In both groups of cirrhotic subjects, baseline levels of plasma atrial natriuretic peptide and cGMP excretion were significantly and comparably elevated compared with the normal range for control subjects ingesting the same sodium intake. Despite the marked difference in the natriuretic response to immersion in both responders and non-responders, there was a significant and comparable further elevation of plasma atrial natriuretic peptide and urinary cGMP excretion during immersion, compared with the control day. CONCLUSION: These results suggest that the relative resistance to the natriuretic action of atrial natriuretic peptide in the non-responders compared with the responders is mediated by anti-natriuretic factors acting at a level parallel with or beyond atrial natriuretic peptide release or coupling to its cGMP-linked receptors.