Recovery Of Developed Tension Research Articles

Cardioprotection after acute exposure to simulated high altitude in rats. Role of nitric oxide

AimIn previous studies, upregulation of NOS during acclimatization of rats to sustained hypobaric hypoxia was associated to cardioprotection, evaluated as an increased tolerance of myocardium to hypoxia/reoxygenation. The objective of the present work was to investigate the effect of acute hypobaric hypoxia and the role of endogenous NO concerning cardiac tolerance to hypoxia/reoxygenation under β-adrenergic stimulation. MethodsRats were submitted to 58.7 kPa in a hypopressure chamber for 48 h whereas their normoxic controls remained at 101.3 kPa. By adding NOS substrate L-arg, or blocker L-NNA, isometric mechanical activity of papillary muscles isolated from left ventricle was evaluated at maximal or minimal production of NO, respectively, under β-adrenergic stimulation by isoproterenol, followed by 60/30 min of hypoxia/reoxygenation. Activities of NOS and cytochrome oxidase were evaluated by spectrophotometric methods and expression of HIF1-α and NOS isoforms by western blot. Eosin and hematoxiline staining were used for histological studies. ResultsCytosolic expression of HIF1-α, nNOS and eNOS, and NO production were higher in left ventricle of hypoxic rats. Mitochondrial cytochrome oxidase activity was decreased by hypobaric hypoxia and this effect was reversed by L-NNA. After H/R, recovery of developed tension in papillary muscles from normoxic rats was 51–60% (regardless NO modulation) while in hypobaric hypoxia was 70% ± 3 (L-arg) and 54% ± 1 (L-NNA). Other mechanical parameters showed similar results. Preserved histological architecture was observed only in L-arg papillary muscles of hypoxic rats. ConclusionExposure of rats to hypobaric hypoxia for only 2 days increased NO synthesis leading to cardioprotection.

Bepridil, an Antiarrhythmic Drug, Opens Mitochondrial KATPChannels, Blocks Sarcolemmal KATPChannels, and Confers Cardioprotection

Bepridil, which is clinically useful in the treatment of arrhythmias, has been reported to inhibit sarcolemmal ATP-sensitive K(+) (sarcK(ATP)) channels. However, the effect of bepridil on mitochondrial ATP-sensitive K(+) (mitoK(ATP)) channels remains unclear. The objective of the present study was to determine whether bepridil activates mitoK(ATP) channels and confers cardioprotection. SarcK(ATP) channels composed of Kir6.2+SUR2A in human embryonic kidney (HEK) 293 cells were examined using the patch-clamp technique. Flavoprotein fluorescence in guinea pig ventricular cells and matrix volume in isolated rat heart mitochondria were measured to assay mitoK(ATP) channel activity. Mitochondrial Ca(2+) concentration ([Ca(2+)](m)) was measured by loading cells with rhod-2 fluorescence. Coronary-perfused guinea pig ventricular muscles were subjected to 35-min no-flow ischemia followed by 60-min reperfusion. Bepridil (10 microM) completely inhibited the pinacidil-induced Kir6.2+SUR2A channel current expressed in HEK 293 cells. Bepridil reversibly oxidized the flavoprotein and increased mitochondrial matrix volume in a concentration-dependent manner. Furthermore, bepridil significantly attenuated the ouabain-induced increase of [Ca(2+)](m). Pretreatment with bepridil for 5 min before ischemia improved the recovery of developed tension measured after 60 min of reperfusion. These effects of bepridil were abolished by the mitoK(ATP) channel blocker 5-hydroxydecanoate (500 microM) and by the nonselective K(ATP) channel blocker glisoxepide (10 microM). Our results indicate that bepridil is an opener of mitoK(ATP) channels but an inhibitor of sarcK(ATP) channels and exerts a direct cardioprotective effect on native cardiac myocytes. This is the first report of a unique modulator of K(ATP) channels; bepridil would be expected to mitigate ischemic injury while blunting arrhythmias.

Effect of sustained hypobaric hypoxia during maturation and aging on rat myocardium. I. Mechanical activity

Long-lasting cardioprotection may be attained by chronic hypoxia. The basal parameters of contractile function and their response to hypoxia/reoxygenation were measured under isometric conditions, in papillary muscles isolated from left ventricle of rats that were submitted to 53.8 kPa in a hypobaric chamber from 7 wk of age and for their lifetime and of their siblings kept at 101.3 kPa. During acclimatization, hematocrit increased, body weight gain decreased, and heart weight increased with right ventricle hypertrophy. Papillary muscle cross-sectional area was similar in both control and hypoxic groups up to 45 wk of exposure. Developed tension (DT) was 34-64% higher in rats exposed to hypoxia for 10, 26, and 45 wk than in their age-matched controls, whereas resting tension was unchanged. Maximal rates of contraction and relaxation showed a similar pattern of changes as DT. Recovery of DT and maximal rates of contraction and relaxation after 60-min hypoxia and 30-min reoxygenation was also improved in adult hypoxic rats to values similar to those of young rats. Heart acclimatization was lost after 74 wk of exposure. Results are consistent with the development of cardioprotection during high-altitude acclimatization and provide an experimental model to study the mechanisms involved, which are addressed in the accompanying paper.

Protective effects of cyclopiazonic acid on ischemia-reperfusion injury in rabbit hearts.

The cardioprotective effects on myocardial ischemia of the sarcoplasmic reticulum (SR) Ca2+-ATPase (SERCA) inhibitor, cyclopiazonic acid (CPA), were studied. We used the isolated arterially perfused interventricular septum of the rabbit heart submitted to 30-min global ischemia/30-min reperfusion. Mechanical [maximal increase in resting tension (MIRT), and the recovery of developed tension (RDT)], and biochemical parameters [creatine phosphokinase activity (CPK) in the effluent] were analyzed. CPA, 1 microM, perfused 30 min before the ischemia intervention significantly increased RDT by 54% and lessened MIRT by 66%. CPA also decreased CPK in the perfusate by 67.7 and 71.4% at 0-2 and 5-7 min of reperfusion, respectively. No additional benefits were shown either when the drug was perfused, both during ischemia and reperfusion, or with higher CPA concentrations (10-30 microM). The CPA cardioprotection was lost when the drug was present only during the reperfusion period. CPA exhibits functional and biochemical cardioprotective effects on myocardial ischemia. We postulated a decreased SR calcium contribution to the initial cytoplasmic calcium overload as the most probable mechanism involved.

Class I antiarrhythmic drugs alter the severity of myocardial stunning by modulating ATP-sensitive K+ channels in guinea pig ventricular muscles.

The effects of various class I antiarrhythmic drugs and glibenclamide were examined on the recovery of contraction during reperfusion, in relation to the action potential duration (APD) seen during ischemia. Action potential and contractile tension were recorded from isolated guinea pig right ventricular muscles perfused with oxygenated Tyrode solution via the coronary artery. Ten minutes of no-flow ischemia shortened the APD at 90% of repolarization level (APD90) to 58% of control (pre-ischemic values). The APD90 was completely restored after 60 min of reperfusion. The developed tension was abolished during ischemia and recovered to 87% of control after 60 min of reperfusion. In the presence of Vaughan Williams class Ia drug cibenzoline (5 microM) or an ATP-sensitive potassium (K(ATP)) channel blocker glibenclamide (10 microM), the shortening of the APD90 during ischemia was significantly attenuated. However, the recovery of developed tension was significantly inhibited. Class Ic drug pilsicainide (10 microM) did not affect the ischemia-induced shortening of the APD90 or the recovery of developed tension after reperfusion. In the presence of class Ib drug mexiletine (10 microM), the shortening of the APD90 during ischemia was significantly facilitated. The recovery of developed tension in the presence of mexiletine tended to be improved, although the difference was not statistically significant. The developed tension measured after the 60 min reperfusion period following 20 min of no-flow ischemia was markedly depressed, indicating the presence of myocardial stunning. Mexiletine and pilsicainide significantly improved the recovery of developed tension and diminished the stunning. We conclude that cibenzoline and glibenclamide, which block cardiac K(ATP) channels inhibit contractile recovery after reperfusion by attenuating the shortening of APD during ischemia. In contrast, mexiletine, which activates K(ATP) channels (in addition to blockade of Na+ channels) improves contractile recovery by facilitating the shortening of APD during ischemia.

Postischemic cardiac performance in the insulin-resistant JCR:LA-cp rat

Hearts from hyperinsulinemic, insulin-resistant JCR:LA-cp rats do not properly regulate intracellular Ca2+ concentration. We hypothesized, therefore, that these hearts may be unusually sensitive to ischemic insults in which Ca2+ overload would be expected. We investigated the response to global ischemia of hearts from JCR: LA-cp animals at three different ages. At 3 mo of age, isolated hearts from insulin-resistant cp rats were mildly resistant to both mild and severe ischemic insults in comparison to the lean control rat hearts. However, at 6 and 9 mo of age, the cp rats demonstrated a poorer recovery of developed tension after ischemia and/or a higher level of resting tension during reperfusion than the lean controls. Postischemic glycogen and ATP contents were significantly lower and lactate content was higher in hearts from 6-mo-old cp rats compared with controls. The results demonstrate that an insulin-resistant animal model exhibits an increased sensitivity to ischemic myocardial injury that develops with advancing age. The mechanism responsible for the enhanced sensitivity may involve augmented glycolytic metabolism. The data also emphasize the importance of the type of diabetes when cardiac dysfunction is examined.

Ischemic preconditioning in human and rat ventricle.

The signal transduction of ischemic preconditioning involves activation of endogenous receptor-based systems, including alpha 1-adrenoceptors and adenosine receptors. Whereas preconditioning protects against ischemia-reperfusion injury, it is unknown whether this protective strategy might be useful clinically. Furthermore, human atrium has been successfully preconditioned, but it is unknown whether human ventricle can be functionally protected against hypoxia-reoxygenation. To study these questions, isolated rat ventricle and human ventricular trabeculae were suspended in an organ bath and subjected to 30 min of hypoxia and 60 min of reoxygenation. In the rat ventricle, preconditioning was induced by 5 min of rapid pacing at 3 Hz in hypoxic buffer without glucose (simulated ischemia), alpha 1-adrenoceptor stimulation (phenylephrine), or adenosine receptor stimulation (adenosine). In the human trabeculae the effects of preceding simulated ischemia and alpha 1-adrenoceptor and adenosine receptor stimulation were examined against hypoxia-reoxygenation. In the rat, pretreatment with simulated ischemia and alpha 1-adrenoceptor and adenosine receptor stimulation improved recovery of developed tension (56 +/- 3, 56 +/- 4, and 58 +/- 2%, respectively) compared with control trabeculae (25 +/- 2%) after hypoxia-reoxygenation (P < 0.05). In human trabeculae, simulated ischemic preconditioning and alpha 1-adrenoceptor and adenosine receptor stimulation augmented recovery of developed tension (65 +/- 5, 59 +/- 6, and 60 +/- 3%, respectively) compared with control (29 +/- 2%) after hypoxia-reoxygenation (P < 0.05). We conclude that functional cardioadaptation (preconditioning) against hypoxia-reoxygenation injury in rat and human myocardium exists and that alpha 1-adrenergic and adenosine receptor signaling participate in conferring this protection.

Electrical heterogeneity and conduction block in reoxygenated guinea pig papillary muscles.

To investigate the changes in electrical activity after reoxygenation, guinea pig papillary muscles were reoxygenated with 5 mM glucose solution after various durations (30-120 min) of substrate-free hypoxia. Action potential duration recovered to the control level on reoxygenation following 30-120 min hypoxia. The recovery of developed tension or resting tension was incomplete after reoxygenation following longer periods of hypoxia. There was a high incidence of arrhythmias on reoxygenation after 60 min hypoxia, which have been shown to be triggered activities due to delayed afterdepolarizations. In some muscles reoxygenated after prolonged hypoxia (90-120 min), there was electrical heterogeneity which was shown by variable action potential durations and configurations. There was also conduction block around the microelectrode site in muscle. It was suggested that the electrical heterogeneity and conduction block could predispose to reentry, and that triggered activities and reentry could be involved in the genesis of arrhythmias on reperfusion.

Preconditioning in isolated superfused human muscle

Studies in isolated superfused rabbit papillary muscles indicate that preconditioning (PC) is not confined to arterially perfused myocardium. In the present study PC of isolated human right atrial trabeculae was investigated avoiding the problems of invasive experimentation in patients. Atrial trabeculae were suspended in an organ bath, superfused with Tyrode's solution and field stimulated at 1 Hz. After stabilization, muscles were randomly allocated to five groups (n = 8 per group). Control (C) muscles had no additional treatment. PC was induced by 3 min rapid pacing at 3 Hz with hypoxic substrate-free buffer, followed by reoxygenation with substrate for 12 min. In two additional groups 8-p-sulfophenyltheophylline (SPT) was added to the superfusate either during stabilization in controls (C+SPT) or during preconditioning (PC+SPT). In the final group, R-phenyl-isopropyl adenosine (R-PIA) was added to the superfusate for 5 min to see whether or not this could substitute for preconditioning. All muscles were then exposed to 90 min hypoxia with no substrate and pacing at 3 Hz, followed by 120 min reoxygenation at 1 Hz. Recovery of developed tension was significantly improved by PC 46.5 +/- 2.4% v 24.6 +/- 2.3% in controls) and this protective effect was blocked by the addition of SPT without adversely affecting controls (recovery in PC+SPT, 25.8 +/- 4.1% and C+SPT, 22.7 +/- 2.9%). R-PIA protected the muscles to a similar extent as PC (43.8 +/- 1.9%). These data provide evidence for the involvement of adenosine in preconditioning in human myocardium.

Lack of platelet-activating factor release on acute myocardial ischemia in the isolated interventricular septum of rabbit heart

The effects of platelet-activating factor (PAF) on myocardial injury after 1 h global ischemia-30 min reperfusion were investigated in isolated arterially perfused interventricular septum of rabbit heart. PAF did not significantly affect developed tension, ±d T/d t max, resting tension and the times of active state in non-ischemic septa. The recovery of developed tension was significantly reduced by PAF (100 nM), after an ischemia-reperfusion challenge, from the control value of 20.9 ± 3.5% to 10.5 ± 1.8%, without a change in the resting tension (15.7 ± 2.8 vs. 15.6 ± 1.3 g). BN 52021 (20 μM), alone did not modify either parameter of ischemic damage, but antagonized the aggravating effect of PAF. Evidence of PAF activity was not found in any of the samples of the effluent perfusate obtained from ischemic control experiments. On the basis of the present results, we suggest a direct role for PAF in aggravating the myocardial damage induced by ischemia, and discard heart cells as the source of PAF in this state.

Lack of platelet-activating factor release on acute myocardial ischemia in the isolated interventricular septum of rabbit heart

The effects of platelet-activating factor (PAF) on myocardial injury after 1 h global ischemia-30 min reperfusion were investigated in isolated arterially perfused interventricular septum of rabbit heart. PAF did not significantly affect developed tension, ±d T /d t max , resting tension and the times of active state in non-ischemic septa. The recovery of developed tension was significantly reduced by PAF (100 nM), after an ischemia-reperfusion challenge, from the control value of 20.9 ± 3.5% to 10.5 ± 1.8%, without a change in the resting tension (15.7 ± 2.8 vs. 15.6 ± 1.3 g). BN 52021 (20 μM), alone did not modify either parameter of ischemic damage, but antagonized the aggravating effect of PAF. Evidence of PAF activity was not found in any of the samples of the effluent perfusate obtained from ischemic control experiments. On the basis of the present results, we suggest a direct role for PAF in aggravating the myocardial damage induced by ischemia, and discard heart cells as the source of PAF in this state.

Pretreatment with the adenosine triphosphate-sensitive potassium channel opener nicorandil and improved myocardial protection during high-potassium ca rdioplegic hypoxia

We hypothesized that pretreatment with the potassium channel opener nicorandil might enhance myocardial protection achieved by cold (20 degrees C) high-potassium (16 mmol/L) cardioplegia (5 ml/min) during long-duration (120 minutes) myocardial hypoxia (average oxygen content 5.4 ml/dl). We tested a 15-minute infusion of nicorandil (1 mmol/L) given only before (group A, n = 8) or before and during cardioplegia (group B, n = 8) in guinea pig papillary muscle preparations contracting isometrically while stimulated (4 mA, 2 msec) at 1600 msec cycle length. Nicorandil was significantly negative inotropic before cardioplegia and shortened significantly action potential duration. During cardioplegia, time to arrest of contraction was shortened from 145 +/- 28 seconds (mean +/- standard error) in the vehicle group (dimethyl sulfoxide 1:100; n = 8) to 56 +/- 10 seconds (p < 0.02) and 68 +/- 5 seconds (p < 0.05) in groups A and B, respectively. Recovery of developed tension at 60 minutes of normothermic reoxygenation (expressed as percent of prehypoxia basal value) was ameliorated from 54% +/- 6% (vehicle group) to 92% +/- 4% (group A, p < 0.01) and to 119% +/- 19% (group B, p < 0.01). The specific potassium channel blocker glibenclamide (glib: 1 mumol/L, n = 8) prolonged action potential duration and was without effect on time to arrest. On reoxygenation, the glib group had prolonged time to half relaxation (versus group A, p < 0.02) and the worst percent developed tension at 60 minutes (40% +/- 4%). In the overall study, time to arrest and percent developed tension at 60 minutes were inversely correlated (r = -0.45, p < 0.01). Arrhythmias were never observed. Multivariate analysis showed that pretreatment with nicorandil (with or without drug adjunct to cardioplegic solution) was a significant factor (r2 = 0.65, p = 0.0001) to influence reoxygenation-mediated recovery of mechanical function. Neither the negative inotropic effect of nicorandil before cardioplegia nor its abbreviating action on time to arrest during cardioplegia was contributory to explain recovery of function on reoxygenation. In subgroup analysis, negative inotropism and the shortening of action potential duration were contributory factors. These data suggest that nicorandil pretreatment activates potassium channels and enhances the myocardial protection provided by cold cardioplegia an effect, which is evident after a long hypoxic period, late on reoxygenation.

Antioxidant changes in heart hypertrophy: significance during hypoxia-reoxygenation injury.

Because hypertrophied rat hearts display an increase in antioxidant enzyme activities and because hypoxia-reoxygenation injury is known to involve free radicals, we tested the hypothesis that the hypertrophied heart may be more resistant to this type of injury. Hypertrophied rat hearts after 10 weeks of chronic pressure overload showed elevated superoxide dismutase (SOD) and glutathione peroxidase (GSHPx) activities and a decrease in lipid peroxidation as indicated by malondialdehyde (MDA) content. Glucose-free hypoxia for 15 min resulted in a complete failure of developed tension and about 200% increase in resting tension in both hypertrophied and sham control groups (p < 0.05). Upon reoxygenation for up to 30 min, hypertrophied hearts recovered developed tension to 60% and resting tension was higher by only 80% of prehypoxic values. In contrast, sham hearts showed only a 25% recovery of developed tension, whereas resting tension remained 130% higher than prehypoxic control values. During hypoxia, the SOD activity was significantly reduced in both sham and hypertrophied groups, whereas GSHPx was reduced only in the sham group. Upon reoxygenation there was no further change in these enzyme activities. Both the SOD and GSHPx activities in the hypertrophied group remained significantly higher than the corresponding reoxygenated sham hearts. During hypoxia, there was no apparent change in MDA content in either the sham or hypertrophied hearts. However, reoxygenation resulted in a significant increase in MDA content in both sham and hypertrophied hearts, but the MDA content was significantly less in the hypertrophied group (p < 0.05). It is suggested that maintenance of an adequate endogenous antioxidant reserve during hypoxia may be important in recovery upon reoxygenation.

Protection by benzamil against dysfunction and damage in rat myocardium after calcium depletion and repletion

Perfusion of the rat right ventricular wall muscle for 4 min with a Ca2(+)-free medium followed by perfusion with a Ca2(+)-containing solution resulted in a 42% recovery of developed tension, contracture, and a massive release of creatine phosphokinase (CPK) and lactate dehydrogenase (LDH) from the muscle. High concentrations (1-5 mM) of amiloride partially protected the ventricular wall from Ca2+ paradox-induced dysfunction. The inclusion of benzamil, an amiloride analogue, 2 min before and during the Ca2(+)-free perfusion period prevented contracture development, restored force development, and almost totally eliminated the release of CPK and LDH from the muscle. Contractile function was best protected by 10-50 microM benzamil. The results demonstrate the efficacy of benzamil as a protective agent against Ca2+ paradox-induced myocardial dysfunction and damage. In view of the known capacity of benzamil to block transsarcolemmal Na(+)-Ca2+ exchange, this study supports the involvement of elevated intracellular Na+ and a stimulation of Na(+)-Ca2+ exchange in this model of cardiac pathology.

The role of the sarcoplasmic reticulum in the response of ferret and rat heart muscle to acidosis.

1. The photoprotein aequorin was micro-injected into papillary muscles from the right ventricle of ferrets and rats. Tension and aequorin light (a function of intracellular [Ca2+]) were monitored. 2. In stimulated ferret papillary muscles, increasing the [CO2] of the bicarbonate-buffered superfusate from 5% (pH 7.35) to 20% (pH 6.8) led to a rapid decrease of developed tension, with no significant change in the size of the intracellular Ca2+ transient which accompanies contraction. There was then a small brief recovery of tension which was accompanied by a large brief increase in the size of the Ca2+ transient. Tension then declined again before recovering more slowly, with no significant change in the size of the Ca2+ transient. 3. The time course of the Ca2+ transient was prolonged on exposure to the acid solution, but shortened on continued exposure to the acid solution. Relaxation of twitch tension became faster on exposure to the acid solution, but slowed again on continued exposure to the acid solution. 4. In the presence of 10 mM-caffeine the size of the Ca2+ transient increased during the initial decline of developed tension, the short-lived recovery of tension was abolished, and the Ca2+ transient became smaller during the slower recovery of developed tension. 2 microM-ryanodine had similar effects on developed tension. 5. Addition of 10 mM-lactic acid to the superfusate produced changes similar to those described in 2 and 3 above. 6. An intracellular acidosis, produced by the addition and subsequent withdrawal of 20 mM-NH4Cl from the superfusate also caused changes similar to those described above. In the presence of caffeine, withdrawal of NH4Cl produced changes similar to those described in 4 above. 7. In unstimulated ferret papillary muscles, increasing superfusate [CO2] produced an increase of aequorin light when the bathing [Ca2+] was increased or in the presence of ouabain (10 microM). This increase was not inhibited by verapamil (5 microM), carbonyl cyanide p-trifluoromethoxyphenylhydrazone (1 microM) and oligomycin (2.5 microM), but was reduced by ryanodine (2 microM). 8. Rat papillary muscles showed responses which were quantitatively different from those observed in ferret papillary muscles: the initial recovery of tension developed more slowly, and sarcoplasmic reticulum (s.r.) inhibitors had a greater inhibitory effect on the recovery of tension. 9. It is concluded that the early decline of developed tension observed during acidosis is due to a decrease in Ca2+ release by the s.r. and a decrease in Ca2+ binding by the myofilaments.(ABSTRACT TRUNCATED AT 400 WORDS)

Intracellular pH in sheep Purkinje fibres and ferret papillary muscles during hypoxia and recovery.

1. The changes of intracellular pH (pHi) of papillary muscles from ferret and Purkinje fibres from sheep heart during hypoxia and recovery from hypoxia were recorded with pH-sensitive micro-electrodes filled with neutral H+ carrier. 2. Hypoxia was produced by replacement of O2 with N2 in the superfusing solutions. When oxidative phosphorylation was prevented, developed tension fell within 20 min to about 16 and 21% of its control value for papillary muscle and Purkinje fibres respectively. On restoration of O2, recovery of developed tension in ferret papillary muscle is preceded by a transient additional decrease. 3. In ferret papillary muscle, the pHi first increased by a mean value of 0.11 pH units after 3 min hypoxia, then decreased by about 0.24 pH units after 20 min. In sheep Purkinje fibres, the initial alkalosis was small or absent, and after 5-9 min, the pHi started to fall reaching 0.17 pH units after 20 min of hypoxia. On return to oxygenated solution, a transient additional intracellular acidification occurred. This acidification reached its peak of 0.31 pH units in papillary muscle and of 0.13 pH units in Purkinje fibres. In both preparations hypoxia was accompanied by a depolarization of a few millivolts. 4. The presence of cyanide (1-2 mM) or fluorodinitrobenzene (20-40 microM) prevented the additional intracellular acidification occurring on return to oxygenated solution. Removal of cyanide itself produced a transient but smaller and slower acidification. 5. On both preparations, exposure to a Tyrode solution containing 10 mM-L-lactate produced a transient intracellular acidification. After recovery from this acidification the acidification produced by hypoxia was increased without affecting the extra acidification on reintroduction of O2. 6. After reduction of the rate of glycolysis by removal of glucose and application of 2-deoxyglucose, the transient intracellular acidification, occurring on return to oxygenated solution after hypoxia, was inhibited in both preparations. In ferret papillary muscle, insulin (100 mU/ml) potentiated the changes of pHi occurring during hypoxia. 7. Using Na+-sensitive glass micro-electrodes it was found that the intracellular Na+ activity rose slightly during the later stages of hypoxia and rose transiently on readdition of O2. These results are consistent with a Na+-H+ exchange being stimulated by acidosis. 8. The origins of the pH changes during and after hypoxia are discussed as are the differences between the responses of sheep Purkinje fibres and ferret papillary muscle.

The effects of verapamil, quiescence, and cardioplegia on calcium exchange and mechanical function in ischemic rabbit myocardium.

The effects of verapamil (1 mg/liter, 2 x 10(-6) mol/liter), quiescence, and cardioplegia (K+ 16 mmol/liter, Mg2+ 16 mmol/liter) on calcium exchange and mechanical function during ischemia and reperfusion have been investigated in the rabbit interventricular septum at 32 degrees C. Calcium influx and efflux were recorded continuously with 47Ca2+ and 45Ca2+. After 60 minutes of total ischemia and reperfusion for 30 minutes, there was a net calcium gain of 4.9 mmol/kg dry tissue. Verapamil given before total ischemia reduced net calcium gain to 1.5 mmol/kg dry tissue (n = 5, P less than 0.03). When given only on reperfusion after total ischemia, or 10 minutes before reperfusion during low flow ischemia, verapamil did not affect calcium exchange. Cardioplegia begun 10 minutes before total ischemia reduced net calcium gain to 1.0 +/- 0.26 mmol/kg dry tissue (n = 6, P less than 0.001). Cardioplegia during the first 10 minutes of reperfusion, or lack of electrical stimulation during reperfusion, did not reduce calcium gain. Net calcium gain correlated with the maximum rise in resting tension and with the recovery of developed tension. In control experiments neither verapamil nor cardioplegia altered influx or efflux of slowly exchanging calcium. The cardioprotective effects of cardioplegia and the calcium channel blocker verapamil appear to be due to a reduction of myocardial work rather than to any specific direct action on calcium fluxes across the myocardial cell membrane.