• YAKHAK HOEJI
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• v.30 no.3
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• pp.149-156
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• 1986

Many experiments have showed that the sodium and potassium ion transporting system and the Na, $^+K^+$-ATPase activity of membrane fragments are inhibited by digitalis glycosides and that the pump may be associated with the pharmacological receptor for the drugs. The aim of our investigation is to elucidate the ouabain binding sites occupation in heart following infusion of ouabain to intact animals by the $^3H$-ouabain binding assay. Lethal dose and 26 percent of lethal dose of ouabain were infused to intact rabbit through ear vein. Microsomal fraction was fractionated from ouabain treated rabbit heart. $^3H$-ouabain binding to these fraction in vitro was studied by the Schwartz's method. $^3H$-ouabain binding to heart microsomal fraction was also studied following infusion of ginseng ethanol extract and caffeine to rabbits respectively. 1) The infusion of lethal dose ouabain (113$\mu\textrm{g}$/kg) inhibited the specific $^3H$-ouabain binding to rabbit heart microsomal fraction to the level of 60% (p<0.01) of control group and the infusion of 26% of lethal dose of ouabain led to the level of 79% (p<0.01) of the control group. 2) Time course of binding of 0.4$\mu{M}$ $^3H$-ouabain to microsomal fraction from rabbit heart following infusion of lethal and 26% of lethal dose of ouabain showed dose dependence at various incubation time. 3) Compared with control, only slight change of $K_d$ and $B_{max}$ was detected in in vitro $^3H$-ouabain binding after infusion of ginseng ethanol extract (300mg/kg) to rabbit. 4) In caffeine infusion group, $^3H$-ouabain binding yielded nearly the same results as control group.

• YAKHAK HOEJI
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• v.28 no.3
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• pp.129-138
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• 1984

Specific [$^{3}H$] ouabain binding and $Ca^{2+}$ -uptake were measured to elucidate the role of high affinity [$^{3}H$] ouabain binding site in rat cardiac myocytes which contain 65% of rod cells. High affinity [$^{3}$H] ouabain binding site, which is about 3% of total pump sites, with apparent dissociation constant ($K_{D}$) of $1.1{\times}10^{-7}M$ and maximum binding site concentration (Bmax) of 1.2 pmol/mg protein ($1.754{\times}10^{5}cells$) were identified. At the concentration of $10^{-7}M$ to $10^{-4}M$, ouabain produced concentration dependent increase in $Ca^{2+}$-uptake of myocytes. The effect of ouabain on $Ca^{2+}$-uptake was not effected by membrane depolarization (elevated K+ in incubation medium) or verapamil. These results suggest that in rat ventricular myocytes the ouabain receptor complex to high affinity site may increase Na+ - $Ca^{2+}$ exchange across the sarcolemmal membrane by inhibition of Na+, K+ - ATPase.

• The Korean Journal of Pharmacology
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• v.23 no.1
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• pp.9-14
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• 1987

The effects of acebutolol and carbamazepine on ouabain-induced arrhythmias were investigated in rabbits. Ouabain produced ventricular arrhythmias which persisted for 7-8 min at the mean dose of $69{\pm}1.3\;{\mu}g/kg$. Ouabain arrhythmias were converted to normal sinus rhythm by administration of acebutolol or carbamazepine singly but lower dosages increased the recovery time. And then, ouabain arrhythmias were effectively converted to normal sinus rhythm and prevented by combined administration of carbamazepine and acebutolol. Each of the combined doses was ineffective when given singly. From the above results, it may be concluded that carbamazepine and acebutolol inhibited the ouabain-induced arrhythmias depending on the level of dosage and showed synergistic interaction.

• The Korean Journal of Physiology
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• v.18 no.2
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• pp.139-150
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• 1984

Vanadate is a potent inhibitor of Na-K-ATPase. Ouabain, the another specific inhibitor of Na-K-ATPase, induces the contraction in cardiac muscle and smooth muscle. But, some investigators observed the discrepancies between vanadate and ouabain-induced contraction in cardiac muscle. The difference of vanadate and ouabain-induced contraction was investigated in the cat ileal smooth muscle. The following results were obtained. 1) Ouabain-induced contraction was biphasic, but vanadate-induced contraction had one peak. 2) Atropine inhibited ouabain·induced contraction, but did not inhibit vanadate-induced contraction. 3) Changes in external $Ca^{++}$concentration or $Ca^{++}$ antagonists had a greater influence on the contraction induced hy vanadate than by ouabain. 4) Removal of $Na^+$ from incubation medium and high $K^+$ abolished ouabain-induced contraction, but had no effect on vanadate-induced contraction. 5) Vanadate-induced contraction was potentiated in the presence of ouabain. 6) After 3 hrs incubation with vanadate, there was no change in intracellular $Na^+$ concentrations in contrast with ouabain. These results suggest that vanadate contracts ileal smooth muscle through the mechanism different from ouabain, and this is independent of the inhibition of Na-K-ATPase activity.

• Biomolecules & Therapeutics
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• v.2 no.2
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• pp.114-119
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• 1994

[$^3$H]Ouabain binding parameters ( $K_{D}$ and $B_{max}$) to control rat ventricular strips and Langendorff preparations which were not previously exposed to ouabain were compared with those to both preparations that had been first exposed to a complete ouabain dose range of dose-response curve (10$^{-8}$ to 10$^{4}$M). In rat ventricular strips and Langendorff perfused heart preparations, cumulative dose-response curves of ouabain revealed biphasic positive inotropic effects, a "low-dose" effect and a "high-dose" effect with E $d_{50}$ values of 0.5 $\mu$M and 35 $\mu$M ouabain, respectively. The "low-dose" effect in ventricular strip disappeared or was diminished significantly when the ouabain dose-response curve was repeated after the washout of the effects of the first dose-response curve, whereas there were no significant differences in the maximal "high-dose"effect in both exposures to oubain. However, both of the control and ouabain-preexposed Langendorff perfused hearts revealed the same low-dose effects. The $K_{D}$ value for [$^3$H] ouabain binding and the ouabain binding site concentration ( $B_{max}$) estimated by [$^3$H]ouabain displacement assay in control preparations were 230 nM and 2 pmol/mg protein, respectively. [$^3$H]Ouabain binding parameters were not changed by repeated exposure to high concentrations of ouabain. These results suggest that digitalis receptor desensitization in the rat ventricular strip may due to the change of post-receptor events induced by ouabain binding to a high affinity site ($\alpha$$_2$isoform).).).).).

• The Korean Journal of Pharmacology
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• v.6 no.1
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• pp.1-7
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• 1970

The effects of ouabain and diphenylhydantoin on ATPase activity in rat erythrocyte membranes were studied and also influence of K on ATPase activity was studied. The ATPase activity of rat erythrocyte membrane has been shown to consist of two components. The first component requires the Mg but occurs in the absence of Na or K (Mg-ATPase) and is not inhibited by ouabain and stimulated by diphenylhydantoin. The second component requires the presence of Mg and also Na or K (Na-K-Mg-ATPase). It is inhibited by ouabain and is stimulated by diphenylhydantoin in low Na concentration and inhibited in high Na concentration. K inhibit Na-K-Mg-ATPase which is inhibited by ouabain. Ouabain and diphenylhydantoin show reversed effect to Na-K-Mg-ATPase activity. It suggest that the therapeutic effect of diphenylhydantoin on digitalis induced cardiac arrhythmia may be resulted from their effect on ion transport mechanism of cell membrance. And the relevance of these findings to the action of ouabain and diphenylhydantoin on membrane transport mechanism is discussed.

• The Korean Journal of Pharmacology
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• v.12 no.1
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• pp.31-44
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• 1976

It has been reported that many of the effects of digitalis glycosides could be mediated partly through the central nervous system. In this study the effects of ouabain given directly into the lateral ventricle of the brain on the renal function of the rabbit were investigated. Intraventricular ouabain elicited antidiuresis in doses ranging from 0.1 to $3\;{\mu}g$, exhibiting a rough dose-response relationship, and decreased the renal plasma flow, glomerular filtration rate and urinary excretion of sodium and potassium, concomitant with the decrease of urine flow. These decreases in urine flow, excretory rate of electrolytes significantly correlated with the decrease in renal plasma flow or glomerular filtration rate, suggesting that the antidiuresis might have been induced by the hemodynamic changes. Intravenous ouabain in a dose of $1\;{\mu}g$ did not affect the renal function. Systemic blood pressure as well as cardiac activity was not affected by the intraventricular ouabain. Effects of the intraventricular ouabain on renal function were abolished by the intravenous phentolamine-pretreatment but not affected by intraventricular phentolamine-pretreatment. Neither vasopressin infusion nor hydration did affect the renal effects of intraventricular ouabain. From these observations, it is suggested that the antidiuresis of intraventricular ouabain is induced by the increased sympathetic influence to the kidney.

• The Korean Journal of Physiology
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• v.19 no.2
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• pp.203-213
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• 1985

This study was carried out to investigate whether the K-pNPPase activity in renal cortical slices can be used as an index for measuring the activity of $Na^+-K^+$ exchange pump. The K-pNPPase activity, Ouabain-sensitive oxygen consumption add intracellular electrolytes content in slices, and Na-K-ATPase activity in microsome were measured and the effects of ouabain and vanadate on these were observed. The results are as follows : 1) p-NPPase activity in slices increased linearly with incubation time during 60 minutes, and $K^+$-dependent, ouabain-sensitive fraction was about 55% of total p-NPPase activity. This value was almost the same through out the incubation time. 2) The concentrations of ouabain and vanadate for 50f inhibition of K-pNPPase activity were$7.0{\times}10^{-6}M$ and $1.3{\times}10^{-5}M$, respectively. 3) The ouabain-sensitive oxygen consumption in slices was reduced to 50% of control value by $6.3{\times}10^{-6}M$ ouabain or $2.5{\times}10^{-5}M$ vanadate. These concentrations were similar to those for 50% inhibition of K-pNPPase activity. 4) The trends of intracellular electrolytes change by ouabain and vanadate were similar to those of the change in K-pNPPase activity. 5) The Na-K-ATPase activity in microsome prepared from renal cortex was completely inhibited by $10^{-3}M$ ouabain or $10^{-3}M$ vanadate and the concentration for 50% inhibition was $1.2{\times}10^{-6}M$ in ouabain and $1.6{\times}10^{-6}M$ in vanadate, which were much lower than those for K-pNPPase activity or ouabain-sensitive oxygen consumption in slices. These results indicate that K-pNPPase activity measured in renal cortical slices is a better index for evaluating $Na^+-K^+$ exchange pump activity than Na-K-ATPase activity measured in microsome.

• Journal of Aquaculture
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• v.11 no.2
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• pp.279-282
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• 1998

Effects of high concentration of intracellular calcium on estradiol-induced vitellogenin(VTG) induction were examined using ouabain in Primary hepatocyte culture in the rainbow trout Oncorhynchus mykiss. Ouabain increases cytosolic free calcium as a result of inhibition of $Na^+ - Ca^{2+}$ exchanger. Ouabain markedly reduced VTG production to the control level, despite of calcium concentrations in the incubatin medium. Therefore, ouabain would reduce VTG production not by increasing intracellular calcium bt directly by inhibiting $Na^+ - K^+$ ATPase.

• Proceedings of the Korean Society of Applied Pharmacology
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• 1994.04a
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• pp.301-301
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• 1994

〔$^3$H〕Ouabain binding parameters(K$\_$D/ and B$\_$max/,) in homogenates prepared fpom control rat ventricular strip and Langendorff preparations which were not previously exposed to ouabain were compared to those in homogenates from ventricular strip and Langendorff preparations that had been first exposed to a complete ouabain dose-response curve(10$\^$-7/M to 10$\^$-4/ M). In rat ventricular strips and Langendorff perfused rat heart preparations, cumulative dose-response cruves of ouabain revealed biphasic positive inotropic effects, a "low-dose" and a "high-dose" effect with ED$\_$50/ values of 0.5${\mu}$M and 35${\mu}$M ouabain, respectively- The "low-dose" effect in rat ventricular strips disappeared or was diminished significantly when the ouabain dose-response curve wag repeated after the washout of the effects of the first curve, whereas the maximal "high-dose" effect was identical in both exposures to oubain. However, there was no change in the "low-dose" effects in both sets of the Langendorff perfused hearts. The contractile activity of the pre-exposed strips did not indicate the presence of residual ouabain since their basal contractile force was decreased 10% compared to initial control. 〔$^3$H〕Ouabain binding parameters, K$\_$D/ and B$\_$max/, were not changed comparing homogenate of control ventricular strips with that of strips pre-exposed to ouabain. These results suggest that digitalis receptor desensitization in the rat ventricular strip may due to the change of post-receptor events induced by ouabain binding to a high affinity site(${\alpha}$$_2$ isoform).