Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: DrugBank:EXPT00514 (Amiloride)
 1,513 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

The extracellular pH (pHe) in many solid tumors is often lower than in normal tissues. Cells may survive conditions of acid pHe because antiports in their membrane exchange Na+ for H+, or HCO3- for Cl-, and thus regulate the intracellular pH (pHi). We have therefore assessed the effects of drugs which interfere with regulation of pHi on survival of Chinese hamster ovary and human bladder cancer MGH-U1 cells in tissue culture. Nigericin, an ionophore which acidifies the cytoplasm when cells are placed in medium at low pHe, was not toxic at pHe 6.5 or above but became very toxic as pHe was reduced below this value. Amiloride and 4,4'-diisothiocyanostilbene 2,2-disulfonic acid, inhibitors of the Na+/H+ and HCO3-/Cl- exchangers, respectively, decreased pHi in the presence of nigericin at low pHe. These drugs showed little or no toxicity in the pHe range of 6.0-7.0 but added greatly to the toxicity of nigericin. A combination of all three drugs led to toxicity in the pHe range of 6.5-6.8, well within the measured range of tumor pH, but not at pHe 7.0 or above. A combination of low pH and hypoxia, two conditions likely to be found in regions distant from tumor blood vessels, caused cell mortality in the absence of drugs, and this effect was increased by nigericin used alone or in combination with amiloride and 4,4'-diisothiocyanostilbene 2,2-disulfonic acid. These drugs may be regarded as prototypes for potential new anticancer agents that might achieve selective killing of tumor cells by interfering with the regulation of intracellular pH.
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PMID:Cytotoxicity of compounds that interfere with the regulation of intracellular pH: a potential new class of anticancer drugs. 381 51

In these studies, we have tested the hypothesis that bile acid-dependent bile formation is attributable, in part, to the stimulation of active bicarbonate secretion and have further explored the cellular mechanism(s) possibly involved in this process using the isolated perfused rat liver. Under control conditions, ursodeoxycholic acid (UDCA) infusion (3 mumol/min X 20 min) produced a 3.7-fold increase in bile flow and a 7.4-fold increase in HCO3- output. Amiloride (an inhibitor of Na+-H+ exchange) decreased UDCA-stimulated bile flow by 20.6% and decreased biliary HCO3- output by 24.9% but increased biliary UDCA output by 42.9%. Thus amiloride decreased UDCA choleretic efficiency (microliter UDCA-stimulated bile/mumol UDCA output) by 45% and UDCA-stimulated increase in HCO3- output per unit UDCA secreted by 48%. Substitution of Li+ for Na+ in perfusate virtually abolished (greater than 95% decrease) both the UDCA choleresis and increase in biliary HCO3- output but modestly decreased (39.6%) biliary bile acid output. Li+ substitution thus decreased UDCA choleretic efficiency by 98% and the UDCA-stimulated increase in HCO3- output by 96%. Amiloride had no effect and Li+ substitution produced a modest decrease in basal bile flow (26.0%) and HCO-3 output (33.5%). Neither amiloride nor Li+ substitution significantly affected UDCA uptake by cultured hepatocytes or by perfused liver. Amiloride (1 mM) also decreased taurocholate (TC)-stimulated choleresis by 48.5%, biliary TC output by 7.2%, and the choleretic efficiency of TC by 45%.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:Effects of Na+ replacement and amiloride on ursodeoxycholic acid-stimulated choleresis and biliary bicarbonate secretion. 382 45

Isolated submandibular glands of adult rats were perfused through the arterial system with oxygenated, HCO3-containing or HCO3-free physiological salt solutions. Secretion of saliva was then induced with acetylcholine (10(-6) M) in the absence or presence of the ion-transport inhibitors 4,4'-diisothiocyano-2,2'-stilbene disulphonic acid (DIDS), furosemide or amiloride. In HCO3-containing perfusates, 10(-4) M DIDS enhanced the initial secretory response (maximum rate of flow increased 18 per cent), but reduced the overall volume of saliva secreted in a 60-min period by 47 per cent. Furosemide (10(-3) M) alone reduced the volume of saliva by 73 per cent and, in combination with 10(-4) M DIDS, by 82 per cent. Amiloride (10(-3) M) caused a 68 per cent reduction in salivary volumes. Replacement of perfusate HCO3 with HEPES did not affect acetylcholine-induced secretion but enhanced the effects of the transport inhibitors, so that total volume of secretion was reduced 94 per cent by furosemide, 55 per cent by DIDS and 80 per cent by amiloride. In HCO3-containing perfusates, DIDS caused a 30-50 per cent increase in salivary Na+ and residual anion (Na + K - Cl) concentrations but amiloride induced a marked increase in salivary Na+ and Cl- concentrations and a decrease in salivary K+ concentrations. Furosemide caused a marked decrease in salivary Cl- concentrations and a marked increase in residual anions. These effects were similar but of smaller magnitude in HCO3-free, HEPES-containing perfusates.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:Effects of 4,4'-diisothiocyano-2,2'-stilbene disulphonic acid and amiloride on salivary secretion by isolated, perfused rat submandibular glands. 386 69

Secretion by the parotid gland of Na-replete and -depleted sheep was investigated by examining the effects of modifiers of ionic transfer on salivary composition and flow rate. These agents were infused into the arterial blood supply of the vascularly isolated gland in anesthetized sheep. Ouabain inhibited Na+-K+ exchange in the ducts caused by Na depletion and restored the [Na+], [K+], and osmolality to close to those of Na-replete saliva. Ouabain also inhibited Cl- -HCO3- exchange in the ducts in Na repletion and depletion. Amiloride partially inhibited Na+-K+ exchange in Na depletion without affecting Cl- -HCO3- exchange. Monensin potentiated Na+-K+ exchange in Na repletion and depletion. Amiloride and monensin gained access to the saliva, but furosemide and ethacrynic acid were almost totally excluded, and, up to 10(-3) M in blood, they did not affect salivary composition or flow rate. Methazolamide gained free access to saliva but was without effect. 4-Acetamido-4'-isothiocyanostilbene-2,2'-disulfonic acid at 10(-3) M slightly increased salivary [Na+] and [HPO4(2-)]. The results indicate potent effects of ouabain on basolateral Na+-K+ pumps and of amiloride and monensin on transcellular delivery of Na+ to these pumps, but ouabain had no effect on salivary flow rate until O2 consumption approached zero and secretion failed. The findings do not support a proposal that the salivary secretion depends on a Cl- -dependent furosemide-sensitive system energized by Na+-K+-ATPase-dependent Na pumps.
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PMID:Effects of ouabain, amiloride, monensin, and other agents on ovine parotid secretion. 395 28

The purpose of this study was to establish the existence of Na/H exchange in cardiac muscle and to evaluate the contribution of Na/H exchange to pHi regulation. The kinetics of pHi changes in cultured chick heart cells were monitored microfluorometrically with 6-carboxyfluorescein and correlated with Nai content changes analyzed by atomic absorption spectrophotometry; transmembrane H+ movements were evaluated under pH stat conditions. After induction of an intracellular acid load by pretreatment with NH4Cl, a regulatory cytoplasmic alkalinization occurred with a t1/2 of 2.9 min. pHi regulation required external Na+ and was concomitant with transmembrane H+ extrusion as well as a rapid rise in Nai content in an Na/H ratio of 1:1. Microelectrode recordings of membrane potential demonstrated directly the electroneutral character of pHi regulation. Acid-induced net Na+ uptake could be either stimulated by further decreasing pHi or inhibited by decreasing pHo; Na+ uptake was unaffected by tetrodotoxin (10 micrograms/ml), quinidine (10(-3) M), DIDS (10(-4) M), Clo-free solution, or HCO3-free solution. Amiloride (10(-3) M) maximally inhibited both pHi regulation and Na+ uptake; the ID50 for amiloride inhibition of Na+ uptake was 3 microM. Nao-dependent H+ extrusion showed half-maximal activation at 15 mM Nao; Li+, but not K+ or choline+, could substitute for Na+ to support H+ extrusion. Cao-free solution also stimulated acid-induced Na+ uptake. We conclude that pHi regulation following an acid load in cardiac muscle cells is by an amiloride-sensitive, electroneutral Na/H exchange. Stimulation of Na/H exchange up to 54 pmol/cm2 X s indicates the rapidity of this exchange across cardiac cell membranes. Na/H exchange may also participate in steady state maintenance of pHi.
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PMID:Na/H exchange in cultured chick heart cells. pHi regulation. 396 33

Necturus gallbladder epithelium transports sodium and chloride by a process that first involves the cellular entry of each ion across the apical membrane in an electrically silent process. In this paper we present results from cell volume and fluid flux measurements in the presence of different inhibitors and at normal and reduced sodium concentrations, which bear on the process by which ionic entry is effected. We find that reduction of mucosal sodium to a concentration of 10 mM has no effect on either cell volume or on the rate of transepithelial fluid transport, whereas the complete removal of sodium causes a significant decrease in cell volume in addition to its known inhibitory effect on fluid transport. Amiloride had no effect on cell volume at normal sodium concentrations but markedly reduced it when the sodium concentration was reduced to 10 mM. Amiloride, bumetanide, and dipyridamole markedly and reversibly inhibited fluid transport. Finally, the addition of ouabain to the serosal medium induced cell swelling, which was prevented by the removal of potassium from the mucosal medium. These results indicate that the process of sodium entry at the apical membrane is complicated and likely includes both cotransport (NaCl or Na-K-2Cl) and parallel exchange (Na-H and Cl-HCO3) transport mechanisms, and that the proportion of NaCl transported by the different mechanisms varies with the conditions.
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PMID:Effects of mucosal sodium removal on cell volume in Necturus gallbladder epithelium. 403 72

Micropuncture of bovine lens epithelial cells cultured on plastic culture dishes gave values for the plasma membrane voltage (V) which remained stable for periods of up to several hours. The value of V was mainly in the range -30 to -45 mV, mean value -36.9 +/- 0.5 mV (S.E.M., n = 188). Raising extracellular [K+] from 5 to 40 mM depolarized V by 10 +/- 3 mV. The extent of this depolarization increased with increasing steady-state V. Barium (2 mM) caused a rapid, reversible depolarization of 7.9 +/- 1.2 mV. In the presence of Ba2+, the response to 40 mM K was reduced to 3.6 +/- 1.1 mV. Ouabain (10(-5) M) caused a fast depolarization by 5.3 +/- 1.2 mV. Exposure to calcium-free EGTA-Ringer's depolarized V reversibly by 19.5 +/- 5.0 mV. In Ca-free medium, the depolarization induced by 40 mM K was reduced to 3.2 +/- 2.4 mV. Whereas in control Ringer's sodium conductance (as measured by exposure to a 10 mM [Na]-Ringer's) is small as compared to potassium conductance, it increased markedly in Ca-depleted medium. Amiloride (10(-4) and 10(-3) M) had no effect on this Na conductance. An increase in the relative conductance for sodium was also elicited by Ba2+ (2 mM). Extracellular acidification led to a depolarization, alkalinization to a hyperpolarization. The extent of this effect was virtually equal in the absence or presence of HCO3-, excluding a significant pathway for bicarbonate. No evidence for a significant chloride conductance could be obtained.
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PMID:Response of the intracellular potentials of cultured bovine lens cells to ions and inhibitors. 406 42

We have tested the hypothesis that the rapid stimulation of Na+/H+ exchange by epidermal growth factor (EGF) is a requirement for induction of mitogenesis. BALB/c 3T3 cells exposed for 4 hr at 37 degrees C to both EGF at 1 ng/ml and either 0.2-1 mM amiloride (an inhibitor of Na+/H+ exchange) or 10 microM MK-685 (an amiloride analog and more potent inhibitor of Na+/H+ exchange) incorporated no less [methyl-3H]thymidine during a 1-hr pulse 20 hr later than did cells exposed for 4 hr to EGF alone. Control experiments utilizing low external pH (to dissociate EGF from its receptor) and anti-EGF antibodies indicated that the failure of amiloride to inhibit mitogenesis when copresent with EGF during the first 4 hr was not due to incomplete removal of EGF and complete removal of amiloride at t4. Cells incubated with 200 microM amiloride for 24 hr showed nearly complete inhibition of stimulation by EGF. In comparison, cells incubated with 10 microM MK-685 for 24 hr showed only a slight inhibition of stimulation by EGF. Incubations with amiloride or MK-685 for shorter periods of time indicated that only amiloride inhibited mitogenesis and that this inhibition happened between 4 (t4) and 10(t10) hr after EGF addition, during which time increases in RNA and protein synthesis (required for mitogenesis) occurred. Amiloride inhibited both RNA and protein syntheses in intact cells during this prereplicative period, while MK-685 was without effect. We conclude that (i) inhibition of EGF-induced mitogenesis by amiloride is due not to inhibition of EGF-stimulated Na+/H+ exchange but rather to inhibition of necessary events occurring during the hours immediately prior to the onset of DNA synthesis, these events probably being RNA and protein synthesis and (ii) in cell culture medium buffered with CO2/HCO3-, complete inhibition of EGF-stimulated Na+/H+ exchange does not inhibit EGF-induced mitogenesis and, thus, stimulation of Na+/H+ exchange is not necessary for induction of mitogenesis by EGF.
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PMID:Inhibition of epidermal growth factor-induced mitogenesis by amiloride and an analog: evidence against a requirement for Na+/H+ exchange. 620 56

Changes in pH alter the oscillatory pattern of glucose-induced electrical activity of mouse islet B cells, thereby supporting the hypothesis that changes in intracellular pH (pHi) resulting from glucose metabolism serve as a coupling factor between metabolic and cationic events. A decrease in pHi in the present of 11.1 mM glucose induces an increase in the duration of the active phase similar to that evoked by higher concentrations of glucose. Regulation of pHi appears to occur by Na:H and HCO3:Cl exchange in the plasma membrane, because inhibition by 0.1 mM amiloride and 0.5 mM 4,4'-diisothiocyano-2,2'-stilbene disulfonic acid (DIDS), respectively, induces constant spike activity in the presence of 11.1 mM glucose. If the pH coupling hypothesis is correct, then inhibition of the putative pH regulatory mechanisms and the subsequent decrease in pHi should elicit electrical activity in the presence of subthreshold glucose (less than 7.0 mM). Amiloride induced electrical activity (threshold) at 4.4 +/- 0.3 mM (mean +/- SEM) glucose. The threshold for DIDS was 5.4 +/- 0.2 glucose, whereas with glucose alone the threshold was achieved at 7.0 +/- 0.4 mM. Thus the generation of H+ by glucose may trigger changes in ionic conductances that induce the typical electrical response. Amiloride was found to elicit a secretory response at subthreshold glucose (4.2-7.0 mM) in perifused rat islets. This indicates that pH-induced changes in the ionic events in the B cells also play an important role in information transfer to the secretory complex.
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PMID:Role of pH as a transduction device in triggering electrical and secretory responses in islet B cells. 632 97

Regulation of intracellular pH is an essential function and may be especially significant in the B-cell in which the influence of glucose on electrical activity is modulated by alterations in pH. Two possible regulatory processes have been examined: Na/H and HCO3/Cl exchange, by using inhibitors, an ionophore, and changes of ionic concentrations. In the presence of 11.1 mM glucose we found that DIDS, an inhibitor of anion exchange, elicited a dose-response increase in the relative duration of the active phase with an ED50 of 99 microM. Probenecid (0.5 mM), an inhibitor of anion fluxes, also augmented the electrical activity (EA) due to glucose. Withdrawal of HCO-3 elicited constant spike activity followed by a resumption of burst activity with a greater duration of the active phase compared to control. These data are consistent with predicted cellular acidification. However, reduction of Cl-o by isethionate substitution produced no marked effect on EA. In contrast, SO-4- substitution for Cl- resulted in variable effects characterized by constant spike activity or a decrease in the duration of the active and silent phases along with silent hyperpolarization. Tributyltin, a Cl/OH, ionophore enhanced EA at 0.25 microM with 120 mM Cl-o, but reduced EA with 10 mM Cl- as would be predicted with either cellular acidification or alkalinization, respectively. Amiloride at 100 microM elicited constant spike activity perhaps due to inhibition of Na/H exchange. Reduction of Na+o from 142.8 to 40.8 mM had a similar effect and enhanced the influence of amiloride. It appears therefore that interference with putative pH regulatory mechanisms in the B-cell are consistent with the hypothesis that cell pH is involved in regulation of EA.
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PMID:pH modulation of glucose-induced electrical activity in B-cells: involvement of Na/H and HCO3/Cl antiporters. 634 84


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