RESUMO
Isosymmetric structural phase transition (IPT, type 0), in which there are no changes in the occupation of Wyckoff positions, the number of atoms in the unit cell, and the space group symmetry, is relatively uncommon. Chlorothiazide, a diuretic agent with a secondary function as an antihypertensive, has been proven to undergo pressure-induced IPT of Form I to Form II at 4.2 GPa. For that reason, it has been chosen as a model compound in this study to determine if IPT can be predicted in silico using periodic DFT calculations. The transformation of Form II into Form I, occurring under decompression, was observed in geometry optimization calculations. However, the reverse transition was not detected, although the calculated differences in the DFT energies and thermodynamic parameters indicated that Form II should be more stable at increased pressure. Finally, the IPT was successfully simulated using ab initio molecular dynamics calculations.
Assuntos
Clorotiazida/farmacologia , Teoria da Densidade Funcional , Algoritmos , Anti-Hipertensivos/farmacologia , Simulação por Computador , Diuréticos/farmacologia , Modelos Teóricos , Simulação de Dinâmica Molecular , Transição de Fase , Pressão , Teoria Quântica , TermodinâmicaRESUMO
Calbindin-D28k (CBD-28k) is a calcium binding protein located in the distal convoluted tubule (DCT) and plays an important role in active calcium transport in the kidney. Loop and thiazide diuretics affect renal Ca and Mg handling: both cause Mg wasting, but have opposite effects on Ca excretion as loop diuretics increase, but thiazides decrease, Ca excretion. To understand the role of CBD-28k in renal Ca and Mg handling in response to diuretics treatment, we investigated renal Ca and Mg excretion and gene expression of DCT Ca and Mg transport molecules in wild-type (WT) and CBD-28k knockout (KO) mice. Mice were treated with chlorothiazide (CTZ; 50 mg · kg(-1) · day(-1)) or furosemide (FSM; 30 mg · kg(-1) · day(-1)) for 3 days. To avoid volume depletion, salt was supplemented in the drinking water. Urine Ca excretion was reduced in WT, but not in KO mice, by CTZ. FSM induced similar hypercalciuria in both groups. DCT Ca transport molecules, including transient receptor potential vanilloid 5 (TRPV5), TRPV6, and CBD-9k, were upregulated by CTZ and FSM in WT, but not in KO mice. Urine Mg excretion was increased and transient receptor potential subfamily M, member 6 (TRPM6) was upregulated by both CTZ and FSM in WT and KO mice. In conclusion, CBD-28k plays an important role in gene expression of DCT Ca, but not Mg, transport molecules, which may be related to its being a Ca, but not a Mg, intracellular sensor. The lack of upregulation of DCT Ca transport molecules by thiazides in the KO mice indicates that the DCT Ca transport system is critical for Ca conservation by thiazides.
Assuntos
Calbindina 1/metabolismo , Cálcio/metabolismo , Clorotiazida/farmacologia , Furosemida/farmacologia , Túbulos Renais Distais/efeitos dos fármacos , Magnésio/metabolismo , Eliminação Renal/efeitos dos fármacos , Inibidores de Simportadores de Cloreto de Sódio/farmacologia , Inibidores de Simportadores de Cloreto de Sódio e Potássio/farmacologia , Animais , Western Blotting , Calbindina 1/deficiência , Calbindina 1/genética , Cálcio/urina , Canais de Cálcio/genética , Canais de Cálcio/metabolismo , Imunofluorescência , Regulação da Expressão Gênica , Genótipo , Túbulos Renais Distais/metabolismo , Magnésio/urina , Masculino , Camundongos Endogâmicos C57BL , Camundongos Knockout , Fenótipo , Reação em Cadeia da Polimerase em Tempo Real , Proteína G de Ligação ao Cálcio S100/genética , Proteína G de Ligação ao Cálcio S100/metabolismo , Canais de Cátion TRPV/genética , Canais de Cátion TRPV/metabolismoRESUMO
The main distal nephron segment sodium transporters are the distal tubule chlorothiazide-sensitive sodium chloride cotransporter (NCC) and the collecting duct amiloride-sensitive epithelial sodium channel (ENaC). The infusion of ghrelin into the renal interstitium stimulates distal nephron-dependent sodium reabsorption in normal rats, but the mechanism is unknown. Here we localize renal ghrelin receptors (GR) to the cortical collecting duct (CCD). Ghrelin significantly increased phosphorylated serum/glucocorticoid-regulated kinase-1 (pSGK1), a major upstream signaling intermediate regulating ENaC. To test whether increased apical membrane αENaC induced the antinatriuresis, ghrelin was infused in the presence of acute and chronic amiloride, a selective inhibitor of ENaC. In the presence of amiloride, renal interstitial ghrelin failed to reduce urine sodium excretion, suggesting that ghrelin-induced sodium reabsorption is dependent on intact ENaC activity. While the main sodium transporter of the CCD is ENaC, NCC is also present. In response to renal interstitial ghrelin infusion, neither total nor phosphorylated NCC levels are altered. Ghrelin-induced sodium reabsorption persisted in the presence of chlorothiazide (selective inhibitor of NCC), indicating that intact NCC activity is not necessary for ghrelin-induced antinatriuresis. Finally, renal interstitial ghrelin infusion significantly increased interstitial cAMP levels and adenylyl cyclase blockade abolished ghrelin-induced antinatriuresis. Thus, GRs expressed in the CCD regulate sodium reabsorption by cAMP-induced trafficking of ENaC to the apical membrane.
Assuntos
AMP Cíclico/fisiologia , Canais Epiteliais de Sódio/fisiologia , Túbulos Renais Coletores/fisiologia , Receptores de Grelina/fisiologia , Transdução de Sinais/fisiologia , Sódio/metabolismo , Amilorida/farmacologia , Animais , Pressão Sanguínea/efeitos dos fármacos , Pressão Sanguínea/fisiologia , Clorotiazida/farmacologia , Feminino , Grelina/farmacologia , Proteínas Imediatamente Precoces/fisiologia , Modelos Animais , Proteínas Serina-Treonina Quinases/fisiologia , Ratos , Ratos Sprague-Dawley , Transdução de Sinais/efeitos dos fármacosRESUMO
We report that the loop diuretic drugs bumetanide and furosemide used in the treatment of hypertension are GPR35 agonists. We utilized calcium flux, inositol phosphate accumulation, and dynamic redistribution assays to examine the pharmacology of these compounds on the human, mouse and rat GPR35. While potent on human GPR35, neither bumetanide nor furosemide were active against mouse or rat GPR35. Furthermore, the Na(+)-Cl(-) cotransporter inhibi- tors chlorothiazide and hydrochlorothiazide were inactive against GPR35 in all three species. We also demonstrate that GPR35 is expressed in human skin where it has been shown that loop diuretics inhibit histamine-induced flare and itch response. These findings suggest that GPR35 may play an important role in skin cell biology and be a potential target for the treatment of a variety of immune disorders.
Assuntos
Bumetanida/farmacologia , Diuréticos/farmacologia , Furosemida/farmacologia , Receptores Acoplados a Proteínas G/agonistas , Animais , Células CHO , Cálcio/metabolismo , Clorotiazida/farmacologia , Cricetinae , Células HEK293 , Células HT29 , Humanos , Hidroclorotiazida/farmacologia , Fosfatos de Inositol/metabolismo , Camundongos , Ratos , Receptores Acoplados a Proteínas G/biossíntese , Pele/metabolismo , Transfecção/métodosRESUMO
Ethacrynic acid, a potent inhibitor of sodium reabsorption in the ascending limb of Henle's loop, produces a sharp rise in renal venous renin activity within 5 min after intravenous administration in anesthetized dogs. This response persists when volume depletion is prevented by returning urinary outflow to the femoral vein. Comparable studies with chlorothiazide, a diuretic with little or no effect on the medullary portion of the ascending limb of the loop of Henle, failed to produce a significant increase in renal venous renin activity.When administered during ureteral occlusion, ethacrynic acid produced no change in renal venous renin activity until ureteral occlusion was released and flow restored. Following release of the ureters, a prompt rise in renal venous renin was again observed within 5 min of release. Control studies of ureteral occlusion yielded a fall in renal venous renin activity following release of the ureter without administration of ethacrynic acid. These studies identify a prompt stimulatory effect of ethacrynic acid on renin release that is unrelated to volume depletion but dependent upon the presence of tubular urine flow. Although further definition of the site and characteristics of the distal tubular mechanism for stimulation of renin release requires more direct study, the data presented here indicate that changes in sodium concentration in distal tubular fluid serve as a stimulus for renin release.
Assuntos
Ácido Etacrínico/farmacologia , Túbulos Renais/metabolismo , Renina/metabolismo , Sódio/metabolismo , Obstrução Ureteral/metabolismo , Angiotensina II/metabolismo , Animais , Transporte Biológico Ativo , Clorotiazida/farmacologia , Cães , Feminino , Sistema Justaglomerular/enzimologia , Natriurese/efeitos dos fármacos , Veias Renais , Renina/sangue , Cateterismo UrinárioRESUMO
We performed studies on dogs under hydrated conditions, utilizing the rate of free water formation (C(H2O)) as an index of the rate of distal tubular sodium transport. Since C(H2O) could be progressively increased with no evidence of a maximal rate during loading with hypotonic (2.5%) mannitol, it was concluded that there is no limit on distal tubular sodium transport during mannitol loading. In contrast, during hypotonic (0.45%) saline loading C(H2O) rose initially, but as urine flow (V) exceeded 25% of the filtered load C(H2O) attained maximal levels (up to 20% of the filtered load) and remained stable as V increased to 50% of the filtered load. It was concluded that saline loading progressively inhibits proximal sodium reabsorption. Initially, the distal tubule absorbes a large fraction of the proximal rejectate and sodium excretion rises slightly. Eventually, an alteration in distal sodium transport appears which culminates in a maximal rate or transport limit. This distal transport limit provoked by saline loading could not be characterized by a classical Tm as seen with glucose and does not seem to be consequent to high rates of flow through the distal tubule. Regardless of the precise nature of this limit, the major increment in sodium excretion develops during saline loading only after saline alters the capacity of the distal tubule to transport sodium.
Assuntos
Túbulos Renais/efeitos dos fármacos , Manitol/farmacologia , Cloreto de Sódio/farmacologia , Sódio/metabolismo , Equilíbrio Hidroeletrolítico/efeitos dos fármacos , Acetazolamida/farmacologia , Animais , Clorotiazida/farmacologia , Cães , Taxa de Filtração GlomerularRESUMO
The acute effects of chlorothiazide (CTZ) on total (TSCA) and ionized (SCA-plus 2) serum calcium concentrations were studied in three groups of people: (a) eight subjects with normal parathyroid function; (b) six patients with hypoparathyroidism; and (c) two patients with hyperparathyroidism. Most subjects were studied on four occasions; at least 3 days intervened between studies on an individual subject. During each experiment the subject received an i.v. influsion of 5% dextrose in water at 1 ml/min from 8 a.m. to 4 p.m. Additions to the infusions were (a) none; (b) CTZ to deliver 3.33 mg/kg/h; (c) parathyroid extract to deliver 1 U/kg/h; or (d) both CTZ and parathyroid extract at the rates previously indicated. CTZ, when used, was added to the infusion at 10 a.m., parathyroid extract at 8 a.m. When CTZ was infused, the diuretic-induced losses of Na and water were replaced by i.v. infusion. In normal subjects 2 h after the start of CTZ infusion, there was a transient increase in SCA-plus 2 which coincided in time of day with a transient decrease in SCA-plus 2 in control experiments. At that time of day SCA-plus 2 was 4.18 plus or minus 0.12 mg/100 ml in control experiments and 4.56 plus or minus 0.08 in experiments with CTZ, P smaller than 0.025. The corresponding values for (TSCA) were 9.32 plus or minus 0.15 and 9.80 plus or minus 0.30, P smaller than 0.01. Such differences were not observed in the group with hypoparathyroidism. In the two patients with hyperparathyroidism, CTZ produced sustained increases in TSCA and SCA-plus 2. In normal subjects and those with hypoparathyroidism, CTZ plus parathyroid extract infusion resulted in sustained increases in both SCA-plus 2 and TSCA throughout the periods of observation when compared to experiments in which only parathyroid extract was infused, P smaller than 0.01 in all instances. The results suggest that the acute hypercalcemic action of CTZ requires the presence of circulating parathyroid hormone.
Assuntos
Cálcio/sangue , Clorotiazida/farmacologia , Doenças das Paratireoides/metabolismo , Hormônio Paratireóideo/farmacologia , Adulto , Cálcio/urina , Feminino , Humanos , Hiperparatireoidismo/sangue , Hiperparatireoidismo/urina , Hipoparatireoidismo/sangue , Hipoparatireoidismo/urina , Infusões Parenterais , Magnésio/urina , Masculino , Pessoa de Meia-Idade , Doenças das Paratireoides/sangue , Doenças das Paratireoides/urina , Fósforo/urina , Sódio/administração & dosagem , Sódio/urina , Água/administração & dosagemRESUMO
Recently it has been observed that Ca excretion in laboratory rats does not follow a Gaussian distribution, with approximately 10% of them excreting Ca at a rate of 2 SD above the group mean. This phenomenon has been described as spontaneous hypercalciuria (SH). Our studies were designed to define its mechanism. 48 Wistar rats were subjected to metabolic studies to identify SH, prospectively defined as Ca excretion 2 SD above the group mean during 7 d of dietary Ca deprivation (=0.03% by analysis), in the absence of hypercalcemia, PO(4) depletion, or exaggerated natriuresis. Progenies from SH rats were found to have significantly higher urine Ca/creatinine (micrograms per milligram) (male = 38 vs. 23, P < 0.05; female = 79 vs. 60, P < 0.005) with 7/20 males and 9/26 females having values 2 SD above the means of normal. After a 12-h fast and during 10% volume expansion with saline, clearance and micropuncture studies were performed on three groups of acutely parathyroidectomized female rats; (a) normocalciuric (N) progenies from the normal, (b) normocalciuric (NC) progenies from SH, and (c) hypercalciuric (HC) progenies from SH rats. Among these groups, there was no significant difference in body weights, glomerular filtration rate, plasma ultrafiltrable Ca (4.5, 4.6 vs. 4.7 mg/100 g), PO(4), and the fractional excretion (FE) of Na or FE(PO4). FE Ca was significantly higher in HC rats (13.9%) than N (10.1%) and NC (10.7%). Segmental reabsorption of fluid and Na was comparable among the three groups. Fractional delivery (FD) of Ca was, however, significantly increased in the late proximal tubule of HC rats (62 vs. 49 and 46%, P < 0.05). The increased FDCa was no longer apparent in early or late distal tubule (6.9 vs. 6.9 and 7.6%, P = NS). Although FECa exceeded late distal FDCa in all three groups, the increment was significantly greater in HC rats (7.02%) than both N (3.4, P < 0.05) and NC rats (3.05, P < 0.02). The effects of chlorothiazide (27.5 mg/kg/d, i.p. x 7 d) were evaluated in the female offsprings of the SH rats. Before chlorothiazide, average urine Ca/creatinine (253 vs. 77.2) and cyclic AMP (26.6 vs. 13.4 mumol/mg creatinine, P < 0.001) on days 7 and 8 of the Ca-deprived diet were higher than the normal. On days 6 and 7 of chlorothiazide, average cyclic AMP (cAMP) excretion fell to normal range (11.7 vs. 12.7 mumol/mg creatinine) as Ca excretion was reduced to normal (62 vs. 59.4 mug Ca/mg creatinine). WE CONCLUDE: (a) SH, as defined in this study, is an inheritable biochemical marker and renal in origin. (b) The hypercalciuria is independent of parathyroid hormone, changes in plasma Ca and tubular handling of Na. (c) As studied in the PTX and volume expanded conditions of our experiments, decreased Ca reabsorption in superficial proximal convoluted tubule is demonstrable, but the hypercalciuria is probably mediated by diminished Ca transport by the deep nephron. The unlikely possibility of increased secretion by the terminal nephron, however, remains to be excluded. (d) In normal rats, there is internephron heterogeneity in regard to Ca transport during saline loading.
Assuntos
Cálcio/urina , Nefropatias/urina , Túbulos Renais/metabolismo , Animais , Clorotiazida/farmacologia , Creatinina/urina , AMP Cíclico/urina , Feminino , Nefropatias/genética , Nefropatias/metabolismo , Masculino , Néfrons/metabolismo , Hormônio Paratireóideo/sangue , Fósforo/urina , Ratos , Ratos Endogâmicos , Sódio/metabolismoRESUMO
The present studies were designed to further investigate the possibility of heterogeneity of nephron function during Ringer loading in the rat, and to determine the specific nephron segment responsible for this finding. As in previous studies from this laboratory with smaller rats (50-125 g), net addition of sodium between late distal tubule and papillary base (6.9 vs. 10.4% of the filtered load, respectively, P <0.005) was found in more mature rats (170-230 g). In contrast, there was net reabsorption of sodium between these two segments in nonvolume-expanded animals, 1.70 vs. 0.45% of the filtered sodium load, P <0.005. Because nephron heterogeneity of sodium transport during extracellular volume expansion is the most likely explanation for these findings, further studies were performed to determine the specific juxtamedullary nephron segment responsible for the net addition pattern between late distal tubule and papillary base in Ringer-loaded animals. First, a comparison was made of sodium delivery to the late proximal tubule of superficial nephrons vs. the delivery rate to the bend of Henle's loop of juxtamedullary nephrons in both hydropenia and Ringer loading. Fractional sodium delivery was quite comparable between the superficial and juxtamedullary nephrons in both hydropenia and Ringer loading although the absolute level was much greater in both groups of nephrons in the Ringer studies. Chlorothiazide (15 mg/kg loading and 15 mg/kg per h) given during Ringer loading markedly increased late distal sodium delivery, 19% of the filtered load, but did not prevent net addition of sodium at the papillary base. In contrast, furosemide (5 mg/kg loading and 5/mg/kg per h) given during Ringer loading completely reversed the segmental pattern, 35.5 and 28.8% at late distal tubule and papillary base, respectively, P <0.005. These studies demonstrate that the net addition of sodium between late distal tubule and papillary base during Ringer loading is not limited to immature rats and that the segmental pattern does not occur in non-volume-expanded animals. Further, the reversal of the net addition pattern with furosemide, but not chlorothiazide, and the comparable proximal nephron delivery rates in Ringer loading suggest that the loop of Henle of juxtamedullary nephrons reabsorbs less sodium than the same portion of superficial nephrons in this setting. A model is proposed to explain this finding.
Assuntos
Espaço Extracelular/fisiologia , Rim/metabolismo , Sódio/metabolismo , Animais , Transporte Biológico/efeitos dos fármacos , Clorotiazida/farmacologia , Feminino , Furosemida/farmacologia , Rim/efeitos dos fármacos , Masculino , Néfrons/efeitos dos fármacos , Néfrons/metabolismo , Concentração Osmolar , RatosRESUMO
The acute effects of intravenous (i.v.) probenecid and chlorothiazide on renal urate handling were investigated in paired studies in normal men. Uricosuric responses to these agents were compared in the same subjects, both without and with pyrazinamide (PZA) pretreatment. Assuming that PZA selectively inhibits the tubular secretion of urate and that uricosuric agents act by increasing the excretion of filtered urate, then the uricosuric responses (the increment in urate excretion or clearance) should have been unaffected by PZA. Defined in this manner, however, uricosuric responses to probenecid and chlorothiazide were significantly decreased after PZA pretreatment. In order to determine whether PZA diminished other renal actions of chlorothiazide, changes in sodium and inorganic phosphorus excretion were examined. Chlorothiazide produced equivalent natriuretic and phosphaturic responses after PZA pretreatment, indicating that PZA does not interfere with at least some of the renal actions of chlorothiazide. In separate studies, PZA depressed urate excretion by at least 68% during the maintenance of chlorothiazide-induced natriuresis and phosphaturia, suggesting that chlorothiazide does not diminish the anti-secretory action of PZA. The results suggest that probenecid and chlorothiazide may derive their uricosuric properties by facilitating the excretion of both filtered and secreted urate. Possibly, increased excretion of secreted urate might occur through modulation of urate reabsorption at a site distal to tubular secretion, rather than by the direct acceleration of secretory transport. However, PZA-induced interference with the actions of probenecid and chlorothiazide on renal urate transport mechanisms cannot be excluded as a possible explanation for the present results.
Assuntos
Clorotiazida/farmacologia , Rim/efeitos dos fármacos , Probenecid/farmacologia , Pirazinamida/farmacologia , Ácido Úrico/urina , Adulto , Ácidos Aminoipúricos , Clorotiazida/antagonistas & inibidores , Feminino , Taxa de Filtração Glomerular , Humanos , Inulina , Rim/metabolismo , Masculino , Pessoa de Meia-Idade , Natriurese/efeitos dos fármacos , Fosfatos/urina , Fósforo/sangue , Probenecid/antagonistas & inibidores , Sódio/sangue , Ácido Úrico/sangue , MicçãoRESUMO
Anesthetized dogs receiving an infusion of chlorothiazide and ethacrynic acid were given 600-ml infusions of distilled water or dilute dextrose solutions. The absolute rate of tubular sodium reabsorption was depressed, and the glomerular filtration rate was increased during the water loading, despite the associated decreases in plasma sodium concentration and decreases in the filtered load of sodium. The extent to which fractional sodium reabsorption decreased and the excretion of sodium increased was inversely related to the degree to which the filtered load of sodium was depressed as a result of the decreased plasma sodium concentration. We conclude that, in the presence of the diuretic blockade of distal tubular sodium reabsorption, infusion of water depresses proximal tubular reabsorption of sodium and that these changes are qualitatively similar to those previously observed during infusions of saline. Similar depression of tubular reabsorption of sodium and increased excretion of sodium occurred during water loading in the absence of diuretics in dogs undergoing saline diuresis, which presumably provided a high rate of distal sodium reabsorption before water loading. We suggest that volume expansion with water depresses proximal tubular reabsorption of sodium in a manner qualitatively similar to infusions of saline and that the extent to which sodium excretion is increased during water loading is dependent upon 1) the absolute extent to which proximal reabsorption is depressed, 2) the extent to which the filtered load of sodium is maintained in the presence of a falling concentration of sodium in plasma, and 3) the extent to which increased distal reabsorption compensates for the depressed proximal reabsorption of sodium. Mechanisms are suggested whereby the previously reported inverse relationship between plasma concentration of sodium and over-all tubular reabsorption of sodium may be only apparent, and could be the result of physiologic "glomerulotubular balance" during the specific experimental maneuvers.
Assuntos
Clorotiazida/farmacologia , Desoxicorticosterona/farmacologia , Ácido Etacrínico/farmacologia , Glucose/farmacologia , Túbulos Renais/efeitos dos fármacos , Túbulos Renais/fisiologia , Sódio/metabolismo , Vasopressinas/farmacologia , Animais , Proteínas Sanguíneas , Cães , Feminino , Taxa de Filtração Glomerular , Masculino , Potássio/sangue , Potássio/urina , Sódio/sangue , Sódio/urinaRESUMO
Clearance experiments were performed in female mongrel dogs, either intact or thyro-parathy-roidectomized (T-PTX), under pentobarbital anesthesia, to examine the unusual hypocalciuric property of thiazide diuretics. The relationship between calcium clearance (C(Ca)) and sodium clearance (C(Na)) was determined in normal dogs, C(Ca) = 0.79 C(Na); constant infusion of chlorothiazide (CTZ) to provide drug concentrations in plasma of approximately 40 mug/ml modified this relationship; C(Ca) = 0.30 C(Na) (P < 0.001). The magnitude of the dissociating effect of CTZ on the urinary Ca/Na relationship was found to be most highly correlated with urinary drug concentration. Infusion of CTZ (1 mg/min) into one renal artery caused a unilateral decrease (25%) in C(Ca)/GFR while producing a unilateral increase (80%) in C(Na)/GFR. The same dose of CTZ in T-PTX dogs produced an increase in C(Na)/GFR without causing a change in C(Ca)/GFR. The defective response in T-PTX dogs could be ascribed to poor tubular secretion of the drug; when urinary drug concentrations were elevated in T-PTX dogs to the levels found in intact dogs (by infusing more drug), C(Ca)/GFR fell to an equivalent extent. T-PTX dogs showed substantially lower renal extraction of CTZ (42%) than intact dogs (57%); PTH administration to T-PTX dogs increased extraction toward normal (49%). The defective secretion of CTZ could not be attributed to either a decreased tubular maximum or a decreased renal blood flow.
Assuntos
Distúrbios do Metabolismo do Cálcio/induzido quimicamente , Clorotiazida/farmacologia , Rim/efeitos dos fármacos , Animais , Cálcio/sangue , Cálcio/urina , Clorotiazida/administração & dosagem , Cães , Relação Dose-Resposta a Droga , Feminino , Furosemida/farmacologia , Taxa de Filtração Glomerular , Injeções Intra-Arteriais , Rim/irrigação sanguínea , Capacidade de Concentração Renal/efeitos dos fármacos , Glândulas Paratireoides/fisiologia , Fosfatos/sangue , Fluxo Sanguíneo Regional , Artéria Renal , Sódio/sangue , Sódio/urina , Glândula Tireoide/fisiologiaRESUMO
The saluretic effect of the thiazide diuretics has been attributed to inhibition of sodium reabsorption in the distal nephron of the kidney. Recent micropuncture studies have shown, however, that chlorothiazide administration can also inhibit sodium reabsorption in the proximal convolution. To clarify the site of the saluretic effect of chlorothiazide, these micropuncture studies examined the effect of chlorothiazide on chloride transport in the nephron. The effect of chlorothiazide on chloride transport was studied because chlorothiazide's effectiveness as a saluretic is largely due to its ability to enhance sodium chloride excretion; if only changes in sodium transport are examined, it would be then difficult to determine if sodium as bicarbonate or as chloride is affected, since chlorothiazide can inhibit carbonic anhydrase. One group of rats was studied before and after 15 mg/kg per h chlorothiazide. For comparison, another group of rats was studied before and after 2 mg/kg per h benzolamide, a carbonic anhydrase inhibitor. Fractional chloride delivery from the proximal tubule was similarly increased in both groups from 59.4 to 71.0% by chlorothiazide administration, Pless than 0.0001, and from 54.3 to 68.2% by benzolamide administration, P less than 0.001. The increased delivery very of chloride from the proximal tubule was largely reabsorbed before the early distal tubule as fractional chloride delivery to this site increased only from 5.08 to 7.40% after chlorothiazide administration, P less than 0.001, and from 4.50 to 6.29% after benzolamide administration, P less than 0.01. Benzolamide had no effect on chloride reabsorption in the distal convoluted tubule. However, chlorothiazide administration resulted in a marked decrease in distal tubular chloride reabsorption, the fraction of filtered chloride present at the late distal tubule incresing from 1.24 to 6.25%, P less than 0.001. Fractional chloride excretion in the urine increased from 0.29 to 3.44%, P less than 0.001, after chlorothiazide, but did not change after benzolamide. The influence of chlorothiazide on proximal chloride transport presumably is related to its ability to inhibit renal carbonic anhydrase. However, it is not the effect of chlorothiazide in the proximal convolution but rather its effect in the distal convoluted tubule which is primarily responsible for its ability to be an effective saliuretic.
Assuntos
Cloretos/metabolismo , Clorotiazida/farmacologia , Rim/efeitos dos fármacos , Néfrons/efeitos dos fármacos , Animais , Derivados de Benzeno/farmacologia , Sítios de Ligação , Inibidores da Anidrase Carbônica , Cloretos/sangue , Taxa de Filtração Glomerular , Inulina/sangue , Túbulos Renais Distais/metabolismo , Túbulos Renais Proximais/metabolismo , Masculino , Microinjeções , Ratos , Sódio/metabolismoRESUMO
The separate effects of volume expansion and of increased delivery of sodium on sodium reabsorption in the diluting segment of the distal nephron were studied in man. In six normal subjects during a sustained water diuresis, sodium delivery to the distal nephron was increased without volume expansion by the administration of acetazolamide. In these subjects, free water clearance rose linearly as a function of urine flow. In five patients with complete, central diabetes insipidus, distal sodium delivery was increased by the infusion of hypertonic saline during a sustained water diuresis. In four of these five patients, changes in free water clearance were also observed during hypertonic saline diuresis in the presence of distal blockade of sodium reabsorption with chlorothiazide. At high rates of distal delivery the following observations were made: (a) free water clearance was lower and fractional sodium excretion higher during saline diuresis compared to acetazolamide diuresis; (b) although free water clearance was moderately reduced by chlorothiazide at low rates of urine flow, there was no difference in free water clearance between saline loading alone and saline plus chlorothiazide at high rates of urine flow; and (c) during saline loading free water clearance rose without evidence of a limit when increased distal delivery was accompanied by spontaneous increases in glomerular filtration rate, but tended toward a limit when glomerular filtration rate remained constant. The data indicate that during acute volume expansion with saline, there is a decrease in the fraction of delivered sodium reabsorbed in the distal nephron when compared to the response of the distal nephron to comparable increases in distal sodium delivery in the absence of volume expansion.
Assuntos
Diabetes Insípido/metabolismo , Túbulos Renais/metabolismo , Sódio/metabolismo , Equilíbrio Hidroeletrolítico , Acetazolamida/farmacologia , Clorotiazida/farmacologia , Diurese/efeitos dos fármacos , Taxa de Filtração Glomerular , Humanos , Soluções Hipertônicas/farmacologia , Natriurese/efeitos dos fármacos , Cloreto de Sódio/farmacologiaRESUMO
These experiments were designed to define the renal disposition of pyrazinoic acid in a nonhuman primate that is phylogenetically close to man and to relate this to the effects of pyrazinoate on urate excretion. The renal clearance of pyrazinoate was almost always greater than the simultaneous glomerular filtration rate at plasma concentrations ranging from 1.9 to 960 mug/ml. Some inhibitors of tubular secretion, probenecid, MK-282 (an experimental, potent uricosuric drug), p-aminohippurate, iodopyracet, sulfinpyrazone, and mersalyl, reduced clearances of pyrazinoate to values far below filtration rate. Chlorothiazide, allopurinol, and salicylate did not. The clearance of pyrazinoate was not influenced by changes in urine flow. It is concluded that pyrazinoate is actively secreted and actively reabsorbed. Pyrazinoate had a dual effect on urate excretion. At concentrations in plasma less than 10 mug/ml there was a concentration related fall in urate/inulin clearance ratio, reaching values of 10-20% of control. Over the range of 10-100 mug/ml in plasma, the clearance of urate remained maximally depressed. At higher concentrations of pyrazinoate there was a concentration related increase in urate/inulin clearance ratio such that at pyrazinoate levels above 600 mug/ml a definite uricosuric response was obtained. Prior administration of pyrazinoate to give plasma levels of 20-140 mug/ml completely or almost completely prevented uricosuric responses to probenecid, PAH, chlorothiazide, and sulfinpyrazone. Iodopyracet, mersalyl, salicylate and N-acetyl-4-dibutylsulfamoyl-3-trifluoromethylbenzenesulfonamide (MK-282) retained significant uricosuric action, but the activities were probably less than normal. The results are consistent with a model of urate transport involving high rates of bidirectional transtubular flux.
Assuntos
Pan troglodytes/metabolismo , Pirazinas/urina , Ácido Úrico/urina , Uricosúricos/farmacologia , Alopurinol/farmacologia , Ácidos Aminoipúricos/farmacologia , Animais , Transporte Biológico , Ácidos Carboxílicos/urina , Clorotiazida/farmacologia , Depressão Química , Taxa de Filtração Glomerular , Insulina/urina , Iodoperaceto/farmacologia , Túbulos Renais/efeitos dos fármacos , Masculino , Compostos Organomercúricos/farmacologia , Probenecid/farmacologia , Ligação Proteica , Pirazinas/sangue , Salicilatos/farmacologia , Estimulação Química , Sulfimpirazona/farmacologia , Ácido Úrico/antagonistas & inibidoresRESUMO
Thiazide diuretics inhibit Na+ and stimulate Ca2+ absorption in renal distal convoluted tubules. Experiments were performed on immortalized mouse distal convoluted tubule (MDCT) cells to determine the mechanism underlying the dissociation of sodium from calcium transport and the stimulation of calcium absorption induced by thiazide diuretics. Control rates of 22Na+ uptake averaged 272 +/- 35 nmol min-1 mg protein-1 and were inhibited 40% by chlorothiazide (CTZ, 10(-4) M). Control rates of 36Cl- uptake averaged 340 +/- 50 nmol min-1 mg protein-1 and were inhibited 50% by CTZ. CTZ stimulated 45Ca2+ uptake by 45% from resting levels of 2.86 +/- 0.26 nmol min-1 mg protein-1. Bumetanide (10(-4) M) had no effect on 22Na+, 36Cl-, or 45Ca2+ uptake. Control levels of intracellular calcium activity ([Ca2+]i) averaged 91 +/- 12 nM. CTZ elicited concentration-dependent increases of [Ca2+]i to a maximum of 654 +/- 31 nM at 10(-4) M. CTZ reduced intracellular chloride activity ([Cl-]i), as determined with the chloride-sensitive fluorescent dye 6-methoxy-N-(3-sulfopropyl)quinolinium. The chloride channel blocker 5-nitro-2-(3-phenylpropylamino)benzoic acid (NPPB, 10(-5) M) abolished the effect of CTZ on [Cl-]i. NPPB also blocked CTZ-induced increases of 45Ca2+. Resting membrane voltage, measured in cells loaded with the potential-sensitive dye 3,3'-dihexyloxacarbocyanine iodide [DiOC6(3)], averaged -72 +/- 2 mV. CTZ hyperpolarized cells in a concentration-dependent and reversible manner. At 10(-4) M, CTZ hyperpolarized MDCT cells by 20.4 +/- 7.2 mV. Reduction of extracellular Cl- or addition of NPPB abolished CTZ-induced hyperpolarization. Direct membrane hyperpolarization increased 45Ca2+ uptake whereas depolarization inhibited 45Ca2+ uptake. CTZ-stimulated 45Ca2+ uptake was inhibited by the Ca2+ channel blocker nifedipine (10(-5) M). We conclude that thiazide diuretics block cellular chloride entry mediated by apical membrane NaCl cotransport. Intracellular chloride, which under control conditions is above its equilibrium value, exits the cell through NPPB-sensitive chloride channels. This decrease of intracellular chloride hyperpolarizes MDCT cells and stimulates Ca2+ entry by apical membrane, dihydropyridine-sensitive Ca2+ channels.
Assuntos
Cálcio/metabolismo , Clorotiazida/farmacologia , Túbulos Renais Distais/efeitos dos fármacos , Animais , Transporte Biológico/efeitos dos fármacos , Bumetanida/farmacologia , Células Cultivadas , Canais de Cloreto , Cloretos/metabolismo , Técnicas In Vitro , Potenciais da Membrana/efeitos dos fármacos , Proteínas de Membrana/efeitos dos fármacos , Camundongos , Nifedipino/farmacologia , Nitrobenzoatos/farmacologia , Sódio/metabolismoRESUMO
In order to clarify the mechanisms of thiazide diuretic-induced hypocalciuria, the effect of a thiazide was studied for 7 days in seven patients with hypoparathyroidism on Vitamin D and one on calcium infusion, and seven euparathyroid patients with hypercalciuria. In the control group, calcium excretion (mg/24 hr) fell by 44% from 415 to 232 within 4 days and remained at this level. Plasma total calcium corrected for total protein did not change. In the hypoparathyroid group, calcium excretion fell by 11% from 351 to 311 and then returned to the base line level. Plasma total calcium (mg/100 ml) increased from 10.09 to 10.88, 11.29 and 10.77 at the end of the 2nd, 4th, and 7th day of thiazide administration. In the patient having i.v. calcium and no Vitamin D, neither plasma nor urinary calcium changed significantly. In both groups sodium excretion increased on the first 2 days and fell to or below base line level thereafter. Urinary phosphate, magnesium, and potassium increased, plasma phosphate rose, and magnesium and potassium fell. It is concluded that: (a) The hypocalciuric effect of thiazides requires the presence of parathyroid hormone and is not solely a result of sodium depletion. (b) The hypercalcemic effect of thiazides in hypoparathyroidism is due to increased release of calcium from bone and requires the presence of a pharmacologic dose of Vitamin D. (c) Thiazides enhane the action of parathyroid hormone on bone and kidney; Vitamin D can replace parathyroid hormone in this interaction in bone but not in kidney.
Assuntos
Clorotiazida/farmacologia , Interações Medicamentosas , Hipoparatireoidismo/tratamento farmacológico , Meticlotiazida/farmacologia , Vitamina D/uso terapêutico , Adulto , Idoso , Osso e Ossos/efeitos dos fármacos , Cálcio/sangue , Cálcio/metabolismo , Cálcio/urina , Colecalciferol/uso terapêutico , Ensaios Clínicos como Assunto , Di-Hidrotaquisterol/uso terapêutico , Ergocalciferóis/uso terapêutico , Feminino , Humanos , Túbulos Renais/efeitos dos fármacos , Magnésio/urina , Masculino , Pessoa de Meia-Idade , Natriurese , Hormônio Paratireóideo/fisiologia , Fosfatos/sangue , Fosfatos/urina , Potássio/urina , Cálculos Urinários/tratamento farmacológicoRESUMO
The effect of various diuretics on H+ secretion was studied in the isolated short-circuited urinary bladder of the turtle. Mucosal (urinary) chlorothiazide stimulated H+ secretion promptly, from 1.33 +/- 0.24 to 3.03 +/- 0.25 mueq/h (P less than 0.001). The effect was rapidly reversible upon washout of the drug, H+ returning to control levels, 1.37 +/- 0.26 mueq/h (P less than 0.001). Similar effects were observed with mucosal hydrochlorothiazide and mucosal ethacrynic acid/cysteine. Stimulation of H+ secretion occurred in the presence or the absence of exogenous CO2, in the presence or absence of mucosal Na+ and during inhibition of Na+ transport by ouabain. There was no stimulation of H+ secretion by uncomplexed ethacrynic acid or by mucosal furosemide. The nondiuretic sulfonamide, sulfasoxizole, and the nonsulfonamide buffer, borate, had no effect on H+ SECRETION. These observations indicate that the stimulatory effect of diuretics on H+ secretion is not related to active sodium transport, transepithelial electrical potential, or the buffering capacity of the drugs. Since the transepithelial pH gradient at which active H+ secretion was abolished was identical for chlorothiazide-treated tissues (2.68 pH U) as for control tissues (2.65 pH U, NS), the data suggest that the protonmotive force of the H+ pump was unaffected by the diuretic. This observation, plus the rapid onset and reversibility of the drugs, is consistent with an effect on the mucosal membrane to increase H+ conductance (K). The findings raise the possibility that direct enhancement of renal H+ secretion may play a role in the metabolic alkalosis induced by some diuretics.
Assuntos
Diuréticos/farmacologia , Hidrogênio/urina , Tartarugas/urina , Bexiga Urinária/efeitos dos fármacos , Animais , Boratos/farmacologia , Clorotiazida/farmacologia , Cisteína/farmacologia , Condutividade Elétrica , Ácido Etacrínico/farmacologia , Furosemida/farmacologia , Hidroclorotiazida/farmacologia , Hidrogênio/metabolismo , Concentração de Íons de Hidrogênio , Técnicas In Vitro , Mucosa/efeitos dos fármacos , Sódio/fisiologia , Sulfisoxazol/farmacologia , Fatores de Tempo , Bexiga Urinária/metabolismoRESUMO
A close relationship has been observed between the clearance rates of sodium and calcium under a variety of diuretic conditions. The thiazide diuretics act differently in dissociating the renal tubular reabsorption of sodium and calcium. This phenomenon has been further investigated using recollection micropuncture and clearance techniques in a group of 14 dogs subjected to three consecutive experimental phases: expansion to 3% of body weight (BWt) with Ringer's solution, chlorothiazide infusion at 20 mg/kg/h, and furosemide in a prime of 10 mg/kg/ and a 10 mg/kg/h infusion. Diuretic losses were balanced with infusion of equal volumes of Ringer's solution throughout the experiment. Chlorothiazide increased the fractional excretion (FE) of sodium almost threefold while FE(Ca) was not significantly altered. Furosemide increased FE(Na) and FE(Ca) to an approximately equal, and more marked, degree. This dissociation of sodium and calcium reabsorption after chlorothiazide was also evident in the superficial distal tubule, where (tubule fluid/plasma sodium) (TF/P(Na)) increased from 0.32 to 0.49 (P < 0.01) and TF/(ultrafiltrate)UF(Ca) was unchanged (0.35-0.31). Furosemide markedly reduced the transtubular concentration gradient for both sodium (0.86) and calcium (0.94). TF/P(Inul in) decreased progressively from 3.79 to 2.78 to 2.33 in three phases. In the late proximal tubule, chlorothiazide induced a fall of TF/P(Inul in) from 1.57 to 1.44 (P < 0.01), but the ratio TF/UF(Ca): TF/P(Na) was unchanged. Furosemide had no significant proximal effect. It is concluded that acute administration of chlorothiazide reduces sodium reabsorption in the distal hephron, presumably the cortical diluting segment, without affecting calcium reabsorption.
Assuntos
Cálcio/metabolismo , Clorotiazida/farmacologia , Furosemida/farmacologia , Rim/efeitos dos fármacos , Sódio/metabolismo , Animais , Artérias , Pressão Sanguínea/efeitos dos fármacos , Proteínas Sanguíneas/análise , Cálcio/urina , Diurese/efeitos dos fármacos , Cães , Taxa de Filtração Glomerular , Hematócrito , Túbulos Renais/metabolismo , Túbulos Renais Distais/metabolismo , Túbulos Renais Proximais/metabolismo , Taxa de Depuração Metabólica , Néfrons/efeitos dos fármacos , Punções , Sódio/urina , Cloreto de Sódio/farmacologiaRESUMO
Short (2 weeks) and long (12 weeks) term effects of furosemide and chlorothiazide on blood pressure, plasma renin activity, and uric acid concentration were studied in 69 hypertensive patients. Both treatments caused significant reductions in blood pressure and increases in plasma renin activity and uric acid at 2 and 12 weeks in 6) normal renin patients; there was no difference between the effects of furosemide and that of chlorothiazide. Reduction in blood pressure in eight low renin patients who showed smaller changes in plasma renin activity and uric acid was not significant at 2 weeks but significant after 12 weeks of treatment.