The inotropic effect of cardioactive glycosides in ventricular myocytes requires Na+-Ca2+ exchanger function.

Glycoside-induced cardiac inotropy has traditionally been attributed to direct Na(+)-K(+)-ATPase inhibition, causing increased intracellular [Na(+)] and consequent Ca(2+) gain via the Na(+)-Ca(2+) exchanger (NCX). However, recent studies suggested alternative mechanisms of glycoside-induced inotropy: (1) direct activation of sarcoplasmic reticulum Ca(2+) release channels (ryanodine receptors; RyRs); (2) increased Ca(2+) selectivity of Na(+) channels (slip-mode conductance); and (3) other signal transduction pathways. None of these proposed mechanisms requires NCX or an altered [Na(+)] gradient. Here we tested the ability of ouabain (OUA, 3 microm), digoxin (DIG, 20 microm) or acetylstrophanthidin (ACS, 4 microm) to alter Ca(2+) transients in completely Na(+)-free conditions in intact ferret and cat ventricular myocytes. We also tested whether OUA directly activates RyRs in permeabilized cat myocytes (measuring Ca(2+) sparks by confocal microscopy). In intact ferret myocytes (stimulated at 0.2 Hz), DIG and ACS enhanced Ca(2+) transients and cell shortening during twitches, as expected. However, prior depletion of [Na(+)](i) (in Na(+)-free, Ca(2+)-free solution) and in Na(+)-free solution (replaced by Li(+)) the inotropic effects of DIG and ACS were completely prevented. In voltage-clamped cat myocytes, OUA increased Ca(2+) transients by 48 +/- 4% but OUA had no effect in Na(+)-depleted cells (replaced by N-methyl-d-glucamine). In permeabilized cat myocytes, OUA did not change Ca(2+) spark frequency, amplitude or spatial spread (although spark duration was slightly prolonged). We conclude that the acute inotropic effects of DIG, ACS and OUA (and the effects on RyRs) depend on the presence of Na(+) and a functional NCX in ferret and cat myocytes (rather than alternate Na(+)-independent mechanisms).

Biphasic effect of cardiac glycosides on action potential duration in isolated Purkinje fibers.

Despite the historical use of cardiac glycosides, the data describing the electrophysiological characteristics of this class of drug are not fully clear. The present study reported the biphasic effect of cardiac glycosides, digoxin (1.25 microM) and acetylstrophanthidin (0.15 microM), on action potential duration in isolated Purkinje fibers by the conventional glass microelectrode technique. At the cycle lengths of 990, 690 and 490 msec., action potential duration lengthened within 10 min. and shortened after 10 min. of digoxin and acetylstrophanthidin administration. The biphasic effect was observed at a concentration of 4.0 mM [K(+)]o. However, at a higher [K(+)]o concentration of 5.4 mM, only the shortening effect on action potential duration was recorded. These results suggest that the biphasic effect of cardiac glycosides on action potential duration is related to the concentration of extracellular potassium and is not related to the stimulating cycle lengths.

Regulation of vocal fold transepithelial water fluxes.

Vocal fold hydration is critical to phonation. We hypothesized that the vocal fold generates bidirectional water fluxes, which are regulated by activity of the Na(+)-K(+)- ATPase. Western blots and immunohistochemistry demonstrated the presence of the alpha-subunit Na(+)-K(+)-ATPase in the canine vocal fold (n = 11). Luminal cells, basal and adjacent one to two layers of suprabasal cells within stratified squamous epithelium, were immunopositive, as well as basolateral membranes of submucosal seromucous glands underlying transitional epithelia. Canine (n = 6) and ovine (n = 14) vocal fold mucosae exhibited transepithelial potential differences of 8.1 +/- 2.8 and 9.3 +/- 1.3 mV (lumen negative), respectively. The potential difference and short-circuit current (ovine = 31 +/- 4 microA/cm(2); canine = 41 +/- 10 microA/cm(2)) were substantially reduced by luminal administration of 75 microM acetylstrophanthidin (P < 0.05). Ovine (n = 7) transepithelial water fluxes decreased from 5.1 +/- 0.3 to 4.3 +/- 0.3 microl x min(-1) x cm(-2) from the basal to luminal chamber and from 5.2 +/- 0.2 to 3.9 +/- 0.3 microl x min(-1) x cm(-2) from the luminal to basal chamber by luminal acetylstrophanthidin (P < 0.05). The presence of the Na(+)-K(+)-ATPase in the vocal fold epithelium and the electrolyte transport derived from its activity provide the intrinsic mechanisms to regulate cell volume as well as vocal fold hydration.

Ion transport and regulation of respiratory tract fluid output in dogs.

To investigate the regulation of respiratory tract fluid output (RTFO), we collected the RTFO in an anesthetized canine model after a series of pharmacological interventions (inhibition of Na(+)-K(+)-ATPase or Na(+)-K(+)-2Cl(-) cotransporter, 250 microl) and physiological challenges (ionic and/or osmotic perturbation in airway lumen, 250 microl). Whereas 250 microl of aerosolized 0.9% saline caused a transient increase in RTFO, a 250-microl bumetanide-induced increase in RTFO was evident for 18 min and a 250-microl acetylstrophanthidin-induced increase in RTFO persisted for at least 30 min. Dry air ventilation decreased the responses of RTFO to the saline (sham) and acetylstrophanthidin intervention but not the bumetanide intervention. Delivery of 250 mosmol/kgH(2)O ion-free mannitol (250 microl) caused marked increases in RTFO that were little affected by the administration of acetylstrophanthidin or bumetanide 30 min before these challenges. A 250-microl 550 mosmol/kgH(2)O ion-free mannitol challenge caused a more marked and prolonged increase in RTFO. Thus aerosol delivery of a low dose of a cardiac glycoside or a near-isosmotic, ion-free, impermeant osmolyte solution may be therapeutically useful by increasing the clearance of secretions from the tracheobronchial airways.

Effects of cardiac glycosides on atrial contractile dysfunction after short-term atrial fibrillation.

BACKGROUND: Despite a long history of use in the treatment of paroxysmal atrial fibrillation (AF), the efficacy of cardiac glycosides has not been established. If such drugs are beneficial in this condition, the general view is that the benefit must be related to their inotropic actions. METHODS AND RESULTS: To assess the effects of the rapid-acting cardiac glycoside, acetylstrophanthidin (AS), on AF and AF-induced right atrial (RA) "stunning," RA wall motion (with ultrasonic crystals), RA pressure, and peak first derivative of pressure (dp/dt) (with microtip transducers) were measured before and after 5 min of high-intensity rapid atrial stimulation (10 Hz; 10 mA; 1 ms) and after the cessation of poststimulation AF. Measurements were made in neurally intact and autonomically blockaded dogs both before and after the administration of AS (0.01 mg/kg IV bolus and 0.015 mg/kg/h IV infusion). AS prevented the post-AF reduction in RA peak dp/dt under neurally intact and autonomically blockaded conditions, and it prevented the post-AF increase in the RA end-systolic dimension and the decrease in the percentage of RA systolic shortening with autonomic blockade. AS was beneficial whether or not baseline inotropy was enhanced by AS. The duration of AF following atrial stimulation was the same before and after AS, but when compared to controls, AS treatment appeared to prolong AF. CONCLUSIONS: Cardiac glycosides exert a favorable effect on AF-induced RA stunning, but this action is unrelated to its effects on the duration of AF.

Extracellular detection of delayed afterdepolarization of cardiac fibers using signal averaging technique.

AIM: To detect delayed after depolarizations (DAD) in extracellular electrograms using signal averaging technique. METHODS: DAD were induced by acetylstrophanthidin (0.25 mumol.L-1, n = 9) and resibufogenin (0.52 mumol.L-1, n = 5) in sheep cardiac Purkinje fibers. Intracellular voltage was recorded with a conventional microelectrode, and simultaneous extracellular electrograms were recorded differentially from widely spaced electrodes placed in the tissue bath. Noise of electrograms was reduced using signal averaging technique. RESULTS: Acetylstrophanthidin and resibufogenin both induced DAD in the intracellular recording and extracellular DAD (DAD-E) in the extracellular electrogram in sheep heart Purkinje fibers. Acetylstrophanthidin and resibufogenin induced typical changes in the action potential including decrease in action potential amplitude, resting potential, maximum diastolic potential, and action potential duration. Similar shortening occurred in the "Q-T interval" recorded by the extracellular electrogram. With either acetylstrophanthidin or resibufogenin, shortening of stimulation cycle length from 990 ms to 690 ms reduced the coupling interval between action potential upstroke and peak voltage of the DAD (P < 0.01), and the coupling interval between the "QRS" and DAD-E recorded extracellularly (P < 0.01). CONCLUSION: DAD can be detected using the extracellular electrograms combined with the high resolution, signal averaging technique.

Effect of acetylstrophanthidin on action potential duration and relation with extracellular potassium in sheep isolated Purkinje fibers.

AIM: To study the relation between the effect of acetylstrophanthidin on action potential duration (APD) and the extracellular potassium concentration. METHODS: Effect of acetylstrophanthidin (AS 0.15 mmol.L-1) on APD at different extracellular potassium concentrations was studied at the stimulation cycle lengths of 990 and 690 ms in sheep isolated cardiac Purkinje fibers using the standard microelectrode technique. RESULTS: At [K+]o 4.0 mmol.L-1, the biphasic effect of AS on APD appeared obviously. Both APD50 and APD90 were lengthened within the first 10 min of drug exposure. After 10 min, they were shortened at all pacing cycle lengths. On the other hand, at [K+]o 5.4 mmol.L-1, AS only shortened APD markedly without lengthening effect on it. The biphasic and monophasic effects of AS on APD were found at [K+]o 4.0 mmol.L-1 and 5.4 mmol.L-1, respectively. CONCLUSION: The effect of AS on APD was related to the concentration of [K+]o.

Decreased inotropic but relatively preserved relaxation response to cyclic adenosine monophosphate-dependent agents in myopathic human myocardium.

BACKGROUND: Increased intracellular concentrations of cyclic adenosine monophosphate (AMP) stimulate a positive inotropic and lusitropic response in healthy myocardial tissue. Heart failure results in an attenuated inotropic response to cyclic AMP-dependent agents. This study compares the inotropic versus relaxation response to cyclic AMP-dependent and cyclic AMP-independent agents in myocardium from patients with end-stage heart failure and control patients without heart failure. METHODS AND RESULTS: Fifty-four control and 59 myopathic human ventricular trabeculae, 1 mm or less in diameter were placed in physiologic saline, 2.5 mM Ca2+, and stretched to the length at which maximal isometric force developed at 30 degrees C, 0.33 Hz. Dose-response curves plotted as percentage change from baseline versus concentration of drug were determined for acetylstrophanthidin, isoproterenol, isobutylmethylxanthine, and milrinone. Acetylstrophanthidin, a cyclic AMP-independent agent, showed similar increases in peak tension relative to baseline over the entire dose range tested for both control and myopathic heart muscle; its effect on relaxation of control and failing cardiac muscle was equivalent over the dose range tested. In contrast, the inotropic actions of the cyclic AMP-dependent agents, isoproterenol and the phosphodiesterase inhibitors, were significantly decreased in myopathic muscle compared with control muscle, but effects on relaxation in the control and myopathic groups remained relatively preserved. CONCLUSIONS: A relatively preserved relaxant effect is associated with the cyclic AMP-dependent agents, despite significant diminution of their inotropic effects. Thus, in advanced heart failure, patients may continue to benefit from the lusitropic effects of the cyclic AMP-dependent agents, even when the inotropic effects of these agents are severely attenuated.

Ultra-specific immunoassays for small molecules: roles of wash steps and multiple binding formats.

New immunometric forms of immunoassay are much more flexible to use than competitive-format immunoassays for small molecular analytes. An example of the utility of this flexibility is the ability to wash the capture antibody after it has been exposed to analyte but before addition of the labeled reagent. This simple maneuver has a large impact on the specificity obtained from already highly specific assays. We also show that specificity can be further increased by means of our multiple binding assay approach, in which the final reading reflects analyte binding to two different primary capture monoclonal antibodies.

Action potential duration and force-frequency relationship in isolated rabbit, guinea pig and rat cardiac muscle.

The effect of action potential duration and elevated cytosolic sodium concentration on the force-frequency relationship in isolated rabbit, guinea pig and rat papillary muscle preparations was studied. Shortening of action potential duration in guinea pig and rabbit from 150-200 ms to values characteristic of rat (20-40 ms), using the K(ATP) channel activator levkromakalim (15 mumol.l-1), markedly reduced the force of contraction and converted the positive force-frequency relationship into negative one at longer pacing cycle lengths. This conversion was greatly enhanced in the presence of acetylstrophanthidin (0.2-1 mumol.l-1), an inhibitor of the Na-K pump. Acetylstrophanthidin (1 mumol.l-1) alone, however, had no effect on the force-frequency relationship. Prolongation of action potential duration in rat with inhibitors of cardiac K channels (4-aminopyridine [10 mmol.l-1] plus tetraethylammonium [2 mmol.l-1) increased the force of contraction and abolished the negative force-frequency relationship observed in rat at longer pacing-cycle lengths. It is concluded that both action potential duration and cytosolic sodium concentration are major determinants of the force-frequency relationship in mammalian myocardium.

Evidence for a sodium-dependent potassium conductance in frog myelinated axon.

After blockade of the voltage-dependent potassium conductances by intracellular application of 4-aminopyridine and tetraethylammonium in frog myelinated axons, a set of brief (0.1 ms) intracellular depolarizing pulses or a long (200 ms) depolarizing pulse evoked a train of action potentials. Under both experimental conditions a hyperpolarizing afterpotential appeared (duration 367 ms +/- 34, mean +/- S.E., n = 15). The purpose of this study was to investigate the properties of this hyperpolarizing afterpotential. It was found that the hyperpolarizing afterpotential increases in amplitude with: (1) the number of sodium-dependent action potentials; (2) action potential broadening (following potassium channels blockade); and (3) the level of depolarization during a current step. Application of tetrodotoxin prevented the activation of the hyperpolarizing afterpotential by any of the above stimuli. The hyperpolarizing afterpotential was unaffected by: (1) 8-acetyl-strophanthidin, an agent that poisons the electrogenic pumping in the axon; (2) blocking calcium influx with extracellular 10 mM magnesium or 2 mM manganese; and (3) buffering of the intracellular calcium, using EGTA in the recording microelectrode. Extracellular application of tetraethylammonium, but not 4-aminopyridine, reduced the hyperpolarizing afterpotential. The hyperpolarizing afterpotential reversed at >> -92 mV. Increasing the external potassium concentration from 2 to 10 mM shifted the reversal potential +14.5 mV, indicating that the hyperpolarizing afterpotential is a potassium mediated conductance.(ABSTRACT TRUNCATED AT 250 WORDS)

[Excitatory effect of alpha-adrenoceptor on the transient inward current in sheep myocardium].

Acetyl strophanthidin (AS) 0.05 mumol/L was used to induce a stable model of transient inward current (Iti) of sheep Purkinje fibers. The effect of the alpha-adrenoceptor agonist phenylephrine (PE) 0.3, 1.0 mumol/L on the amplitude and duration of Iti was observed in the presence of propranolol 0.5 mumol/L to block the beta-adrenoceptor. When the preparation was perfused with PE 1.0 mumol/L for 20, 50 min, the amplitude of Iti decreased from control value 12.8 +/- 1.9 nA to respectively 10.7 +/- 1.2 nA (n = 5, P < 0.05) and 9.6 +/- 1.9 nA (n = 5, P < 0.01). The D50 of Iti prolonged correspondingly from control value 145 +/- 24.4 ms to 183.3 +/- 28.1 ms (n = 5, P < 0.05) and 207.5 +/- 34.2 ms (n = 5, P < 0.01). The inhibitory effect of PE on Iti showed a dose- and time-dependent manner. Both the time for arrival of the peak value and for return to baseline of Iti prolonged. This suggests that the kinetics of Iti channel changed in the presence of PE. When the Iti was enhanced by beta- adreceptor agonist isoprenaline (ISO) 1.0 mumol/L, the addition of PE 1.0 mumol/L to the perfusate for 10 min, the inhibitory effect become more pronounced, the amplitude of Iti decreased from 15.6 +/- 3.2 nA to 10.3 +/- 2.2 nA and the D50 of Iti prolonged from 92.5 +/- 14.3 ms to 132.5 +/- 36.0 ms (n = 5, P < 0.01).

[Effects of alpha-adrenoreceptor excitation on the delayed after-depolarization of sheep cardiac Purkinje fibers].

By using acetyl strophanthidin (AS) 0.2 mumol/L, the delayed after-depolarization (DAD) was induced in sheep cardiac Purkinje fibers and recorded with intra-cellular microelectrode. When beta-adrenoceptor was blocked by propranolol 1.0 mumol/L, phenylephrine 1.0 mumol/L increased the amplitude of DAD from 8.1 +/- 2.2 mV to 9.5 +/- 2.8 mV, prolonged the duration of DAD from 240 +/- 47 ms to 273 +/- 47 ms (n = 13, P < 0.01) and increased the up rising velocity of DAD from 0.039 +/- 0.023 V/s to 0.051 +/- 0.026 V/s (n = 13, P < 0.05). The DAD occurred earlier for 30 +/- 47 ms to preceding action potential (n = 13, P < 0.05). When triggered action potentials were induced by norepinephrine 1.0 mumol/L on the basis of DAD, propranolol 1.0 mumol/L could suppress the triggered beats while phentolamine 1.8 mumol/L showed little effect. The above results indicate that excitation of alpha-receptor had only slight augmentation effect on DAD. However, for the triggered activity induced by DAD, the inhibitory effect of beta-blockers are stronger than that of alpha-blockers.

Induction of delayed afterdepolarizations and triggered arrhythmias in isolated Purkinje fibers: comparison of resibufogenin and acetylstrophanthidin.

The purpose of this study was to compare the electrotoxicological effects of resibufogenin (RBG) (n = 14) with acetylstrophanthidin (AS) (n = 14) to induce delayed afterdepolarization (DAD) and triggered activity (TA), and their alteration of the electrophysiological properties in sheep cardiac Purkinje fibers using the extracellular electrograms, signal averaging, and standard microelectrode techniques simultaneously. The results indicated: 1) Lower toxic dose of RBG (0.52 mumol.L-1) and AS (0.25 mumol.L-1) induced intracellular and extracellular DAD (DAD-I and DAD-E) at pacing cycle length of 990 and 690 ms. 2) Higher toxic dose of RBG (2.6 mumol.L-1) and AS (5.0 mumol.L-1) induced DAD and TA, nonsustained or sustained premature action potential and oscillatory potentials; 3) At the beginning period of superfusing the drugs, both RBG and AS caused changes of the electrophysiological characteristics. This study demonstrates that the electro-toxicological characteristics and electrophysiological properties of RBG are similar to that of AS and suggests that RBG belongs to the family of digitalis-like drugs.

Reductions of myocardial Na-K-ATPase activity and ouabain binding sites in heart failure: prevention by nadolol.

To study the changes in myocardial digitalis binding sites in heart failure, we measured myocardial ouabain binding sites, Na-K-adenosinetriphosphatase (ATPase) activity, and ventricular muscle mechanical responses to acetylstrophanthidin in dogs with right-heart failure (RHF) produced by tricuspid avulsion and pulmonary artery constriction. Sham-operated dogs were studied as the control. RHF produced a significant decrease in ouabain binding sites in the right and left ventricular myocardium, which was accompanied by a proportional decrease in Na-K-ATPase activity. However, RHF and sham-operated dogs did not differ in systemic hemodynamic or right ventricular trabeculate muscle isometric contractile responses to acetylstrophanthidin. To determine whether chronic beta-adrenergic stimulation contributed to the development of Na-K-ATPase downregulation, we administered nadolol (40 mg/day) to a separate group of dogs during an early stage of RHF development. Nadolol effectively prevented the reduction of myocardial ouabain binding sites that occurred in RHF. Thus we conclude that myocardial ouabain binding sites and Na-K-ATPase activity are reduced in dogs with experimental heart failure and that these changes probably occur as a result of the attendant heightened sympathetic activity.

Effects of acetylstrophanthidin on baroreflex sensitivity in patients with acute myocardial infarction.

We evaluated the effects of acetylstrophanthidin on baroreflex sensitivity in patients soon after an acute myocardial infarction. Baroreflex control of heart rate is frequently depressed after acute myocardial infarction and few data are available as to the effects of pharmacological intervention on this parameter. The reflex chronotropic response to arterial baroreceptor stimulation was assessed in 29 patients with uncomplicated acute myocardial infarction in control conditions (72-96 h after symptom onset) and 30 min after acetylstrophanthidin administration. To check for spontaneous baroreflex sensitivity variations, 24 patients with the same characteristics were evaluated at the same time intervals before and after a 10-cc bolus of saline placebo. Baroreflex sensitivity was assessed by calculating the regression line relating phenylephrine-induced increases in systolic blood pressure to the attendant changes in RR intervals. Mean baseline baroreflex sensitivity value for the whole study population was 7.4 +/- 4.5 ms/mmHg and was unchanged, 7.0 +/- 4.5 ms/mmHg, after acetylstrophanthidin (P = NS). Mean baroreflex sensitivity values were also comparable dividing patients according to the site of infarction both before and after acetylstrophanthidin. Despite the lack of difference in mean baroreflex sensitivity values between the two studies, at a post hoc analysis an inverse relation was found in the total study population between baseline baroreflex sensitivity values and their changes after acetylstrophanthidin (r = -0.62; P < 0.005). The inverse relation was also evident separately in anterior (r = -0.57; P < 0.05) and in inferior (r = -0.70; P < 0.005) myocardial infarction patients.(ABSTRACT TRUNCATED AT 250 WORDS)

Drug-induced afterdepolarizations and triggered activity occur in a discrete subpopulation of ventricular muscle cells (M cells) in the canine heart: quinidine and digitalis.

INTRODUCTION: Oscillations of membrane potential that attend or follow the cardiac action potential and depend on preceding transmembrane activity for their manifestation are known as afterdepolarizations. Early afterdepolarizations (EADs) interrupt or retard repolarization of the cardiac action potential, whereas delayed afterdepolarizations (DADs) arise after full repolarization. EADs and DADs can give rise to spontaneous action potentials or triggered activity believed to be responsible for a variety of cardiac arrhythmias. Recent studies from our laboratory have highlighted differences in the electrophysiology and pharmacology of three functionally distinct myocardial cell types found in the canine ventricle. Epicardial, M region, and endocardial tissues and cells show distinct, sometimes opposite, responses to a variety of drugs, including those capable of inducing EADs and DADs. METHODS AND RESULTS: In the present study, we used standard microelectrode techniques to examine the pharmacologic response of these cellular subtypes to therapeutic levels of quinidine and toxic levels of digitalis. Quinidine readily produced prominent EADs and EAD-induced triggered activity in tissue preparations from the M region (deep subepicardium), but not in those from epicardium, endocardium, or deep subendocardium of the canine ventricle. Acetylstrophanthidin produced prominent DADs in M cell preparations and subendocardial Purkinje fibers but only minute DADs, if any, in epicardium, endocardium, or deep subendocardium. DAD-induced triggered activity was observed to arise only in Purkinje and M cells and never in myocardial tissues from the epicardial, endocardial, or deep subendocardial regions of the ventricular wall. CONCLUSION: We conclude that EADs, DADs, and triggered activity caused by therapeutic levels of quinidine and toxic levels of digitalis are limited to or much more readily induced in a select population of cells in the deep subepicardial (M cell) region of the canine ventricle in addition to the Purkinje system of the heart.

Afterdepolarizations and triggered activity develop in a select population of cells (M cells) in canine ventricular myocardium: the effects of acetylstrophanthidin and Bay K 8644.

Early afterdepolarizations (EADs) are membrane oscillations that interrupt or retard the repolarization phase of the cardiac action potential, whereas delayed afterdepolarizations (DADs) are oscillations that arise after full repolarization. When EADs and DADs are sufficiently large to depolarize the cell membrane to its voltage threshold, they give rise to triggered action potentials, which are believed to underlie some forms of extrasystolic activity and tachyarrhythmias. EAD- and DAD-induced triggered activity have been described and well characterized in isolated Purkinje fibers exposed to a wide variety of drugs, but are rarely seen in syncytial preparations of ventricular myocardium. These results are inconsistent with those of in vivo studies or experiments involving enzymatically dissociated myocytes. In the present study, we used the cardiotonic agent acetylstrophanthidin (AcS) and the calcium channel agonist Bay K 8644 to provide evidence in support of the hypothesis that induction of prominent EADs, DADs, and triggered activity occurs in a select population of cells in ventricular myocardium. The data indicate that EADs, DADs, and triggered activity produced by digitalis and Bay K 8644 are limited to or more readily induced in the deep subepicardial cell layers of the canine ventricle (M cells). Afterdepolarization-induced triggered activity was never observed in the epicardial or endocardial layers.

Effects of different cardiac steroids on intracellular sodium, inotropy and toxicity in sheep Purkinje fibers.

The purpose of the present study was to examine the differences between cardiac steroids that might underlie the variations in toxic/therapeutic ratios that have been reported to occur in vitro as well as in vivo. We used Na(+)-sensitive microelectrodes to measure changes in intracellular Na+ activity (aiNa) associated with positive inotropic and toxic effects of acetylstrophanthidin (AS) and a semisynthetic agent, actodigin. Measurements of aiNa, twitch tension and transmembrane potential were made in sheep Purkinje fibers stimulated at 0.03, 1 and 2 Hz. Ca(+)+i overload toxicity was indicated by the presence of transient depolarizations (TD). The following results were obtained: 1) at a stimulation frequency of 1 Hz, aiNa was significantly higher at peak tension with AS (13.6 +/- 1.1 mM) than with actodigin (11.0 +/- 0.4 mM, P less than .01), yet TD occurred at the same aiNa (10.9 +/- 0.7 vs. 11.9 +/- 0.7 mM, respectively, N.S.); 2) at frequencies of 1 to 2 Hz, aiNa was lower when TD occurred (10.4 +/- 0.9 mM at 2 Hz) than at peak tension (12.1 +/- 0.8 mM, P less than .05) during exposure to AS, whereas aiNa was the same at peak tension (10.6 +/- 1.1 mM) and when TD occurred (10.5 +/- 1.1 mM, N.S.) during exposure to actodigin; 3) the degree of positive inotropy at a high stimulation frequency (2 Hz) was significantly greater with actodigin (about 12-fold increase in force compared to control) than with AS (about 6-fold increase in force).(ABSTRACT TRUNCATED AT 250 WORDS)