Regulation of renal Na-HCO3 cotransporter: III. Presence and modulation by glucocorticoids in primary cultures of the proximal tubule

Regulation of renal Na-HCO3 cotransporter: III. Presence and modulation by glucocorticoids in primary cultures of the proximal tubule. We looked for the presence of the Na-HCO3 cotransporter in primary cultures of the proximal tubule and examined the modulation of this cotransporter by glucocorticoid hormones. Primary cultures of the proximal tubule of the rabbit have Cl-independent, HCO3-dependent 22Na uptake which is DIDS-sensitive. In addition, in cells loaded with BCECF and perfused with Cl-free solution, removal of Na was associated with a decrease in intracellular pH which returned to normal with re-addition of Na. The pH recovery was not inhibited by EIPA but was sensitive to DIDS. These findings are compatible with existence of Na-HCO3 cotransporter in these cells. We examined the role of glucocorticoids on the activity of the Na-HCO3 cotransporter by culturing proximal tubule cells in the presence of hydrocortisone and when confluence was reached, hydrocortisone was deleted from the medium. In the absence of hydrocortisone, the activity of the cotransporter, measured either isotopically or fluorometrically, was significantly decreased, whereas re-addition of hydrocortisone 10-8 M, restored the activity of the cotransporter to normal levels. The effect of hydrocortisone could not be duplicated by aldosterone, suggesting a glucocorticoid-dependent effect. Dexamethasone, a glucocorticoid without mineralocorticoid activity, stimulated the activity of the cotransporter within physiologic concentrations and this effect was blocked by progesterone. The effect of dexamethasone was time-dependent and was prevented by cycloheximide, a protein synthesis inhibitor. These results demonstrate that primary cultures of the proximal tubule have Na-HCO3 cotransporter activity which is modulated by physiological concentrations of glucocorticoids through a protein synthesis-dependent mechanism

Role of urinary concentrating ability in the generation of toxic papillary necrosis

Role of urinary concentrating ability in the generation of toxic papillary necrosis. We studied the pathogenesis of chemically induced papillary necrosis in six groups of rats. Papillary necrosis was produced by a single injection of 2-bromoethylamine hydrobromide (BEA), 50 mg, i.v.; the animals were followed for 7 to 10 days after the administration of the compound. Following BEA, heterozygous Brattleboro rats developed all the functional and morphologic lesions of papillary necrosis that we previously described in Sprague-Dawley rats. They were unable to maintain sodium balance when dietary sodium was withdrawn. Homozygous Brattleboro rats, on the other hand, developed none of the manifestations of papillary necrosis (that is, animals with central diabetes insipidus were protected completely from the nephrotoxic effects of BEA). They adapted normally to a zero sodium diet. Chronic administration of vasopressin to homozygous Brattleboro rats fully restored the toxic effects of BEA. Lowering urinary concentrating ability by inducing a water diuresis in Sprague-Dawley rats completely protected against BEA-induced papillary necrosis. Decreasing papillary solute concentration by furosemide or increasing urine flow after abrupt withdrawal of vasopressin to homozygous Brattleboro rats did not protect against BEA-induced papillary necrosis. We conclude that the combination, but not either alone, of increased urine flow and decreased papillary solute concentration protects against the development of BEA-induced papillary necrosis.Rôle du pouvoir de concentration urinaire sur l'apparition d'une nécrose papillaire toxique. Nous avons étudié la physiopathologie d'une nécrose papillaire induite chimiquement chez six groupes de rats. La nécrose papillaire était produite par une injection unique de 2-bromoéthylamine hydrobromide (BEA), 50 mg, i.v.; les animaux étaient suivis pendant 7 à 10 jours après administration du produit. Après le BEA, des rats Brattleboro hétérozygotes ont développé toutes les lésions fonctionnelles et morphologiques d'une nécrose papillaire telles que nous les avions déjà décrites chez des rats Sprague-Dawley. Ils n'étaient pas capables d'assurer l'équilibre de la balance sodée lorsque le sodium alimentaire était supprimé. Par ailleurs, des rats Brattleboro homozygotes n'ont développé aucune des manifestations de nécrose papillaire (c'est-à-dire que les animaux ayant un diabète insipide central étaient complètement protégés des effets néphrotoxiques du BEA). Ils s'adaptaient normalement à un régime sans sodium. L'administration chronique de vasopressine à des rats Brattleboro homozygotes a totalement restauré l'effet toxique du BEA. Le fait d'abaisser la capacité de concentration des urines en induisant une diurèse aqueuse dans les rats Sprague-Dawley a complètement protégé contre la nécrose papillaire induite par le BEA. La diminution de la concentration de solutés papillaires par le furosémide ou l'augmentation du débit urinaire après arrêt brutal de la vasopressine chez des rats Brattleboro homozygotes ne protégeait pas contre la nécrose papillaire induite par le BEA. Nous concluons que la combinaison, mais non chaque facteur séparément, d'une augmentation du débit urinaire et d'une diminution de la concentration de solutés papillaires protègent contre le développement d'une nécrose papillaire induite par le BEA

Clinical and pathophysiologic spectrum of acquired distal renal tubular acidosis

Clinical and pathophysiologic spectrum of acquired distal renal tubular acidosis. Urinary acidification was studied in nine patients with hyper-chloremic metabolic acidosis. The aim of this study was to investigate the mechanism(s) of impaired distal acidification by the systematic administration of sodium sulfate and neutral phosphate. No impairment of proximal acidification was apparent because all patients had a fractional bicarbonate excretion below 5% at plasma bicarbonate concentrations above 22 mEq/liter. All patients except two were unable to lower urine pH below 5.5 despite systemic metabolic acidosis. The two patients who lowered urine pH normally were hyperkalemic and had selective aldosterone deficiency. Six patients failed to lower the urine pH below 5.5 with sodium sulfate (6.04 ± 0.16) and were unable to achieve a normal urine minus blood (U-B) Pco2 gradient with neutral phosphate (2.8 ± 3.5mm Hg). Control subjects, the two patients with selective aldosterone deficiency, and the remaining patient lowered the urine pH below 5.5 and increased the U-B Pco2 gradient above 25mm Hg in response to sodium sulfate and neutral phosphate infusion, respectively. The abnormal response to these agents exhibited by six patients strongly suggests that the mechanism of impaired distal acidification was that of secretory failure of the proton pump. The normal response of the remaining three patients indicates that the proton pump was able to secrete hydrogen ions normally under maximal stimulation. This pattern is totally predictable in patients with isolated selective aldosterone deficiency who are also capable of lowering the urine pH normally in the presence of systemic metabolic acidosis. The distinctive acidification pattern of the remaining patient who was also hyperkalemic can be explained on the basis of a voltage-dependent type of distal renal tubular acidosis. This type may be disclosed by the findings of impairment of both hydrogen ion and potassium secretion.Aspects clinique et physiopathologique de l'acidose tubulaire distale acquise. L'acidification urinaire a été étudiée chez neuf malades ayant une acidose métabolique hyperchlorémique. Le but de ce travail était d'étudier le mécanisme de l'altération de l'acidification distale par l'administration de sulfate de sodium et de phosphate neutre. Il n'est pas apparu d'altération de l'acidication proximale puisque tous les malades avaient une excrétion fractionnelle de bicarbonate inférieure à 5% à des concentrations de bicarbonate plasmatique supérieures à 22 mEq/litre. Tous les malades sauf deux étaient incapables d'abaisser leur pH urinaire au dessous de 5,5 malgré l'acidose métabolique. Les deux malades qui abaissaient le pH de l'urine à des valeurs normales étaient hyperkaliémiques et avaient un déficit sélectif d'aldostérone. Six malades n'ont pu abaisser leur pH urinaire en dessous de 5,5 avec le sulfate de sodium (6,04 ± 0,16) et ont été incapables de réaliser un gradient de Pco2 normal urine-sang sous phosphate neutre (2,8 ± 3,5mm Hg). Les sujets contrôles, les deux malades ayant un déficit d'aldostérone et le dernier malade ont abaissé le pH de l'urine au dessous de 5,5 et augmenté le gradient de Pco2 à plus de 25mm Hg en réponse aux administrations de sulfate de sodium et de phosphate neutre, respectivement. La résponse anormale des six malades suggère fortement que le mécanisme de l'altération de l'acidification distale est un défaut de fonctionnement de la pompe à protons. La réponse normale des trois derniers malades indique que la pompe était capable de sécréter des ions hydrogène dans des conditions de stimulation maximales. Cette modalité est prévisible chez les malades qui ont un déficit sélectif et isolé d'aldostérone et qui sont aussi capables d'abaisser le pH de leur urine en présence d'une acidose métabolique systémique. La modalité d'acidification particulière du dernier malade qui était en même temps hyperkaliémique peut être expliquée par un mécanisme dépendant de la différence de potentiel. Cette situation peut être reconnue par la constatation d'un désordre portant à la fois sur la sécrétion de ions hydrogène et celle de potassium

Hyperkalemic renal tubular acidosis: Effect of furosemide in humans and in rats

Hyperkalemic renal tubular acidosis: Effect of furosemide in humans and in rats. Furosemide increases urinary acidification in control subjects and in certain patients with normokalemic or hypokalemic distal renal tubular acidosis (RTA). We studied the effect of furosemide in 14 patients with hyperkalemic distal RTA. In a group of patients with pure selective aldosterone deficiency, furosemide increased net acid and K excretion in a fashion indistinguishable from controls. This effect of furosemide was observed both in the presence and in the absence of acute mineralocorticoid administration. In another group of patients with hyperkalemic distal RTA, furosemide failed to decrease urine pH and to increase net acid excretion despite acute mineralocorticoid administration. Plasma aldosterone was variable in this group in that some patients had appropriate levels of aldosterone for the degree of hyperkalemia, whereas in the other patients the levels were low. The failure of these patients to respond to furosemide, despite pharmacologic doses of mineralocorticoid, suggests that these patients had a defect in H+ secretion other than that attributable to aldosterone deficiency alone. To gain insight into the mechanism whereby furosemide increases urinary acidification, we studied control and amiloride-treated rats pretreated with mineralocorticoid. In response to furosemide, control rats had a significantly lower urine pH and higher net acid and K excretion than that observed in amiloride-treated rats. These data suggest that furosemide increases H+ and K excretion, at least in part, by creating a favorable electric gradient for secretion of these ions since these effects were blunted in presence of inhibition of distal Na transport by amiloride. In contrast, in patients and in rats with selective aldosterone deficiency, furosemide increased H+ and K excretion, suggesting that aldosterone is not necessary for the furosemide to increase excretion of these ions

Cholinergic inhibition of urinary acidification by the turtle bladder

Cholinergic inhibition of urinary acidification by the turtle bladder. The effect of carbachol on urinary acidification by the turtle bladder in vitro was studied. Carbachol inhibited urinary acidification in a dose-dependent fashion, with half maximal inhibition occurring at 4.5 × 10-5 M. The effect of carbachol on urinary acidification could be totally prevented by atropine, indicating that the inhibition is mediated through a muscarinic receptor. Carbachol inhibited hydrogen ion secretion by decreasing the active proton conductance and not by altering the proton motive force. Carbachol failed to increase passive loss of hydrogen ion from the mucosa. Carbachol increased calcium uptake by the turtle bladder; this increase in calcium uptake could be prevented by pretreatment with atropine, pentobarbital, or lanthanum. Pentobarbital or lanthanum blunted the inhibitory effect of carbachol on hydrogen ion secretion. In the presence of low extracellular calcium (0.2 mM), carbachol failed to increase calcium uptake but caused a significant inhibition of hydrogen ion secretion. In the presence of normal calcium concentration, carbachol caused a significant efflux of calcium. These data demonstrate that carbachol inhibits urinary acidification and suggest that the mechanism of this inhibition may be related, at least in part, to changes in cytosolic calcium.Inhibition cholinergique de l'acidification urinaire par la vessie de tortue. L'effet du carbachol sur l'acidification urinaire par la vessie de tortue in vitro a été étudié. Le carbachol inhibe l'acidification urinaire, l'inhibition est dose dépendante et la demi-inhibition maximale est obtenue à 4,5 × 10-5 M. L'effèt du carbachol sur l'acidification urinaire peut être complètement empêché par l'atropine, ce qui indique que l'inhibition a pour médiateur un récepteur muscarinique. Le carbachol inhibe la sécrétion d'ions de hydrogène en diminuant la conductance et non en modifiant la force cinétique. Le carbachol n'a pas augmenté la perte passive d'ions de hydrogène par la muqueuse. Le carbachol a augmenté la captation de calcium par la vessie de tortue. Cette augmentation peut être empêchée par un prétraitement par l'atropine, le pentobarbital ou le lanthane. Le pentobarbital ou le lanthane masquent l'effet inhibiteur du carbachol sur la sécrétion d'ions de hydrogène. En présence d'une concentration extracellulaire de calcium faible (0,2 mM), le carbachol n'a pas augmenté la captation de calcium, mais a déterminé une inhibition significative de la sécrétion d'ions de hydrogène. En présence d'une concentration normale de calcium le carbachol entraîne un efflux significatif de calcium. Ces résultats démontrent que le carbachol inhibe l'acidification urinaire et suggère que le mécanisme de cette inhibition peut être en rapport, au moins partiellement, avec des modifications du calcium cytosolique