RESUMEN
Atrial natriuretic peptide (ANP) and endothelin (ET) are endogenous vasoactive factors that exert potent diuretic and natriuretic actions. We have previously shown that ANP and ET-3 act through an NO pathway to inhibit the sodium-glucose cotransporter (SGLT) in the intestine [Gonzalez Bosc LV, Elustondo PA, Ortiz MC, Vidal NA. Effect of atrial natriuretic peptide on sodium-glucose cotransport in the rat small intestine. Peptides 1997; 18: 1491-5; Gonzalez Bosc LV, Majowicz MP, Ortiz MC, Vidal NA. Effects of endothelin-3 on intestinal ion transport. Peptides 2001; 22: 2069-75.]. Here we address the role of ANP and ET-3 on SGLT activity in renal proximal tubules. In rat renal cortical brush border membranes (BBV), fluorescein isothiocianate (FITC) labeling revealed a specific 72-kD peptide that exhibits increased FITC labeling in the presence of Na+ and D-glucose. Using alpha-14C-methylglucose active uptake, rat BBV were shown to possess SGLT activity with an affinity constant (K(0.5) approximately 2.4 mM) that is consistent with the expression of the low-affinity, high-capacity SGLT2 isoform. SGLT2 activity in these preparations is dramatically inhibited by ANP and ET-3. This inhibition is independent of changes in membrane lipids and is mimicked by the cGMP analogue, 8-Br-cGMP, suggesting the involvement of cGMP/PKG pathways. These results are the first demonstration that both ANP and ET-3 inhibit rat cortical renal SGLT2 activity, and suggest a novel mechanism by which these vasoactive substances modulate hydro-saline balance at the proximal tubular nephron level.
Asunto(s)
Factor Natriurético Atrial/farmacología , Endotelina-3/farmacología , Túbulos Renales Proximales/metabolismo , Proteínas de Transporte de Monosacáridos/antagonistas & inhibidores , Animales , Transporte Biológico/efectos de los fármacos , Membrana Celular/efectos de los fármacos , GMP Cíclico/metabolismo , Túbulos Renales Proximales/efectos de los fármacos , Masculino , Microvellosidades/química , Microvellosidades/efectos de los fármacos , Microvellosidades/metabolismo , Peso Molecular , Proteínas de Transporte de Monosacáridos/metabolismo , Óxido Nítrico/metabolismo , Ratas , Ratas Wistar , Transportador 2 de Sodio-GlucosaRESUMEN
We investigated the effects of endothelin 3 (ET-3) on electrolyte transport in rat small intestine using a voltage clamp technique in Ussing's chamber. ET-3 diminished potential difference (PD) and short circuit current (Isc). ET-3 did not affect PD or Isc in low Na(+) and/or D-glucose-free medium. Phloridzine (an inhibitor of sodium-glucose cotransporter [SGLT1]) pretreatment abolished the effect of ET-3 on Isc. Methylene blue (a soluble guanylate cyclase inhibitor) or N-nitro-L-arginine methyl ester (a NOS inhibitor) pretreatment delayed the effect of ET-3 on PD and Isc. ET-3 enhanced NOS activity on enterocytes and systemic NO production. Then, ET-3 could inhibit SGLT1 with the participation of NO.
Asunto(s)
Endotelina-3/farmacología , Intestino Delgado/efectos de los fármacos , Animales , Intestino Delgado/enzimología , Intestino Delgado/metabolismo , Transporte Iónico , Masculino , NADPH Deshidrogenasa/metabolismo , Nitratos/orina , Óxido Nítrico/fisiología , Nitritos/orina , Ratas , Ratas WistarRESUMEN
The intestinal tract is a target organ for atrial natriuretic peptide (ANP), characterized by various biologic activities, immunoreactivity, as well as specific binding sites for ANP. A review of previous studies reveals that ANP is an important regulator of water and nutrient intake, which acts via multiple signaling pathways including activation of guanylyl cyclase to produce its biologic responses. As a regulator, the peptide locally controls hydrosaline balance and acute systemic effects. Therefore, ANP could also act as a local mediator or paracrine effector of intestinal function.
Asunto(s)
Factor Natriurético Atrial/farmacología , Sistema Digestivo/efectos de los fármacos , Intestinos/efectos de los fármacos , Transporte Biológico/efectos de los fármacos , Enterocitos/efectos de los fármacos , Motilidad Gastrointestinal/efectos de los fármacos , Modelos Biológicos , Transducción de SeñalRESUMEN
The aim of the present study was to determine the relationship between the hypotensive effect of the atrial natriuretic peptide (ANP) and the nitric oxide (NO) pathway. N(G)-nitro-L-arginine methyl ester bolus (L-NAME, 1 mg/kg) reverted the decrease in mean arterial pressure induced by ANP administration (5 microg/kg bolus and 0.2 microg x kg(-1) x min(-1) infusion), and the injection of L-NAME before peptide administration suppressed the ANP hypotensive response. To confirm these findings, a histochemical reaction was used to determine NADPH-diaphorase activity (a NO synthase marker) in the endothelium and smooth muscle of aorta and arterioles of the small and large intestine. ANP increased aorta and arteriole endothelium staining after both in vivo administration and in vitro tissue incubation. In both cases, L-NAME prevented the ANP effect on NADPH-diaphorase activity. Tissues incubated with 8-bromoguanosine 3',5'-cyclic monophosphate mimicked ANP action. In addition, ANP administration increased urinary excretion of NO(x) end products. These findings indicate that ANP increases NO synthesis capability and NO production and suggest that the cGMP pathway may be involved. In conclusion, the NO pathway could be an intercellular messenger in the ANP endothelium-dependent vasorelaxation mechanism.
Asunto(s)
Factor Natriurético Atrial/farmacología , Factor Natriurético Atrial/fisiología , Presión Sanguínea/efectos de los fármacos , Presión Sanguínea/fisiología , Óxido Nítrico/fisiología , Animales , Inhibidores Enzimáticos/farmacología , Masculino , NADPH Deshidrogenasa/metabolismo , NG-Nitroarginina Metil Éster/farmacología , Ratas , Ratas WistarRESUMEN
Histochemical reaction of NADPH-diaphorase (NOS-NADPH-d) was used to identify NO synthesis. A 30-min 0.1 microg microg/kg/min ANP infusion led to about a 10% and 35% increase in small and large intestine enterocytes stain respectively. This increase was abolished by a bolus of 1 mg/kg L-NAME before ANP infusion in small intestine, and partially abolished it in colon. Incubation of small and large intestine with 0.5 microM ANP increased stain at about 20%. In both tissues the preincubation with 0.1 mM L-NAME abolished the ANP effect. Incubation with 0.1 mM 8-Br-cGMP enhanced staining about 70% and 30% in small and large intestine respectively. Our results show that ANP enhances NOS-NADPH-d activity, suggesting that ANP stimulates NO synthase in enterocytes by L-arginine-NO pathway. 8-Br-cGMP mimicked the effect of ANP described above. Therefore, the guanylate cyclase-coupled natriuretic receptors, NPR-A and NPR-B, probably mediate this ANP effect.
Asunto(s)
Factor Natriurético Atrial/farmacología , Mucosa Intestinal/efectos de los fármacos , NADPH Deshidrogenasa/análisis , Óxido Nítrico/biosíntesis , Animales , Colon/citología , Colon/efectos de los fármacos , Colon/enzimología , GMP Cíclico/análogos & derivados , GMP Cíclico/farmacología , Guanilato Ciclasa/metabolismo , Histocitoquímica , Procesamiento de Imagen Asistido por Computador , Mucosa Intestinal/citología , Mucosa Intestinal/enzimología , Intestino Delgado/citología , Intestino Delgado/efectos de los fármacos , Intestino Delgado/enzimología , Isoenzimas , Masculino , NG-Nitroarginina Metil Éster/farmacología , Óxido Nítrico Sintasa , Fotoperiodo , Ratas , Ratas Wistar , Receptores del Factor Natriurético Atrial/metabolismoRESUMEN
The aim of the present study was to elucidate the role of an IV dose of endothelin-3 (ET-3) (5 ng Kg-1 min-1) on mean arterial pressure (MAP), on diuresis and natriuresis in control and in volume expanded anesthetized rats. A systemic infusion of ET-3 in normal rats (Group I) increased MAP and produced a trend of increasing diuresis, without changes in natriuresis. A 10% body weight expansion (Group II) increased diuresis and natriuresis without changes in MAP. The simultaneous infusion of ET-3 and expansion with saline (Group III) resulted in an increase in MAP, an enhanced diuretic response, and a natriuresis of similar magnitude to that observed in Group II. These results suggest that the diuresis produced by a low dose of exogenous ET-3 in control rats, is independent of sodium excretion. Furthermore, the enhanced diuresis caused by ET-3 during expansion is greater than the addition of ET-3 and expansion effects, suggesting that new mechanisms are triggered in order to maintain volume and salt homeostasis in this state.
Asunto(s)
Endotelina-3/fisiología , Espacio Extracelular/metabolismo , Sodio/metabolismo , Agua/metabolismo , Animales , Presión Sanguínea/fisiología , Tamaño de la Célula/fisiología , Diuresis/fisiología , Masculino , Natriuresis/fisiología , Ratas , Ratas WistarRESUMEN
Our purpose was to study the influence of the stimulation of the cerebroventricular system on some mechanisms related to hydrosaline equilibrium and blood pressure regulation. Renal function and blood pressure (group 1) as well as the baroreceptor reflex (group 2) were studied. In group 1, we measured diuresis, natriuresis, creatinine clearance, lithium clearance, and blood pressure in control rats and after stimulation of the cerebroventricular system with 1 M NaCl solution. In group 2, we evaluated the baroreceptor reflex, producing an increase of blood pressure with an injection of phenylephrine to obtain baroreceptor reflex curves--characterized by threshold, point of inflection, heart period range, gain, and systolic pressure corresponding to half the heart period range (SBP50)--in control and experimental rats injected with saline and 1 M NaCl solution, respectively. In group 1 experimental rats, we observed a significant increase in diuresis, natriuresis, blood pressure, and glomerular filtration rate. A substantial increase was also registered in sodium filtered load and reabsorbed sodium in the proximal convoluted tubule and distal nephron. No differences were observed either in fractional proximal tubule or in distal nephron sodium reabsorption. In group 2 experimental rats, mean arterial blood pressure, threshold, point of inflection, and SBP50 were significantly higher than in control rats. By contrast, a decrease in gain and heart period range was observed. No difference was obtained in heart rate. Our results demonstrate that the increase of the natriuresis is due, at least in part, to an increase in sodium filtered load.(ABSTRACT TRUNCATED AT 250 WORDS)