Negative inotropic effect of pinacidil on the rat left atria

Abstract— Cromakalim at 50 μm and pinacidil at 0·1–10 μm had no effect, but pinacidil at 0·1 Mm had a negative inotropic effect on the rat electrically-driven left atria without altering the positive inotropic responses to isoprenaline or phenylephrine alone. Glibenclamide had no effect but 4-aminopyridine, procaine (30 μm ) and tetraethylammonium (3 Mm ) augmented the cardiac stimulation response. The ability of pinacidil to attenuate the cardiac stimulation response was not altered by glibenclamide, 4-aminopyridine or procaine but was prevented by pretreatment with tetraethylammonium. Thus, on the rat left atria, pinacidil has a negative inotropic effect which is unrelated to the opening of ATP-sensitive potassium channels, but may be due to opening the inward rectifying potassium channels.     

The influence of oxidative stress on various inotropic responses in isolated rat left atria

The effects of free radicals, generated by electrolysis of a physiological salt solution, on various inotropic responses to drugs in isolated rat left atria were studied. Evidence for the generation of hydroxyl radicals was obtained from an appropriate fluorimetric assay. The amount of free radicals produced by electrolysis of the medium proved current-dependent. Exposure of isolated rat left atria to the medium which had been subjected to electrolysis caused a current-dependent decrease in contractile force. Oxidative stress, as a result of the electrolysis of the medium, caused altered inotropic responses to extra cellular Ca²⁺ (pD₂ control group: 2.62 ± 0.06 vs. 2.44 ± 0.07 electrolysis group), sodium withdrawal (rise in contractile force control group: 1.73 ± 0.19 mN vs. 0.48 ± 0.21 mN electrolysis group) and lowering of stimulation frequency. The response to isoprenaline was diminished in atria subjected to oxidative stress and led to a rightward shift of the concentration response curves (pD₂ control group: 7.56 ± 0.10 vs. 6.77 ± 0.11 electrolysis group). In addition, the inotropic responses to forskolin (pD₂ control group: 6.17 ± 0.12 vs. < 4.5 electrolysis group) and dibutyryl cAMP (rise in contractile force caused by 1 × 10^–5 M db-cAMP in control group: 2.15 ± 0.01 mN vs. 1.21 ± 0.10 mN electrolysis group) proved blunted as well. Measurement of the adenylyl cyclase activity revealed that free radicals attenuated the basal (by 11.1%) and forskolin stimulated (155.0 ± 5.1 vs. 48.0 ± 1.8 pmol cAMP/mg prot./min for control and electrolysis group respectively) activity of the adenylyl cyclase. DMSO, a well known hydroxyl radical scavenger, was able to abolish the free radical-induced decrease in the response to isoprenaline. Surprisingly, addition of α-adrenoceptor agonists to atria subjected to electrolysis-generated free radicals led to a rapid decrease in contractile force. DMSO was unable to counteract the negative intropic effect of methoxamine in atria subjected to oxidative stress. This negative inotropic response to α-adrenoceptor agonists in atria subjected to electrolysed medium is unlikely to be the direct result of phospholipase C or protein kinase C activation. Angiotensin II (which stimulates PLC as well) did not reduce contractile force and chelerythrine (a PKC inhibitor) was unable to counteract the negative inotropic effect of the adrenoceptor agonists. In addition, the negative inotropic effect of methoxamine proved insensitive to 10^–6 M phentolamine and 10^–5 M doxazosin, which indicates an α-adrenoceptor independent mechanism. From this study we conclude that free radicals alter responses to various inotropic stimuli. These alterations may be the result of injured contractile elements, transporter molecules and molecules involved in signal transduction. Addition of α-adrenoceptor agonists after oxidative stress leads to a α-adrenoceptor, PLC and PKC independent decrease in contractile force. Received: 17 June 1996 / Accepted: 6 November 1996     

Inotropic effect of prostacyclin (PGI2) on isolated rat atria at different contraction frequencies

Summary The effects of a wide range of Prostacyclin (PGI₂) concentrations on the Isometric Developed Tension (IDT) of isolated left auricles driven at different frequencies (0.8, 1.6 or 3.3 Hz) were explored. A comparison of the positive inotropism of PGI₂, norepinephrine (NE), tyramine and phenylephrine (Phenyl) indicated that PGI₂, NE and tyramine enhanced peak tension significantly less at slow (0.8 and 1.6 Hz) than at higher rates (3.3 Hz); whereas Phenyl augmented equally the IDT at all of these three driving frequencies. The positive inotropism evoked by PGI₂ was inhibited by (–)-propranolol and also by a treatment with 6-OHDA. Cocaine, normetanephrine and U-0521, augmented the positive inotropic influence of PGI₂ on atria paced at a slow rate (0.8 Hz) but not at a faster one (3.3 Hz). These results suggest that the action of PGI₂ on atrial contractions is apparently indirect and mediated by the release of tissue catecholamines. In addition the effect of PGI₂ and NE at slow and fast rates appears associated with a different relative relevance of the processes which terminate the action of added sympathomimetic agonists, namely, neuronal or extraneuronal uptake as well as metabolization via COMT. These mechanisms seen to be more prominent at slower driving frequencies and could explain the diminished effect of PGI₂ and NE on slowly paced atria.This work was supported by Grant 6638 from the Consejo Nacional de Investigaciones Científicas y Técnicas de la República Argentina (CONICET)     

Naloxone modifies the inotropic decrease induced by halothane on isolated left atria

1. The present study evaluates the interaction between naloxone and halothane on the left atria. 2. Halothane produced significant decrease in auricular inotropism at concentrations ranging from 0.3 to 2.5 v/v%. 3. Naloxone modified the concentration response curve to halothane. The antagonism between both drugs is consistent with a competitive antagonism since the curve obtained had a slope which did not differ significantly from -1. 4. These results suggest that the negative inotropic effects induced by halothane could be mediated by opioid receptors.     

Influence of hypothyroid status on dopamine-induced positive chronotropic and inotropic effects on isolated rat atria

The involvement of alpha- and beta-adrenoceptors in dopamine (DA)-induced positive chronotropic and inotropic effects was investigated in isolated atria from euthyroid and hypothyroid rats. Propranolol at 3 x 10(-7) M remarkably inhibited the positive chronotropic effect of DA in both euthyroid and hypothyroid rats, and 1 x 10(-6) M phentolamine inhibited the effects of DA in the presence of propranolol in hypothyroid rats. Propranolol remarkably inhibited the positive inotropic effect of DA in both euthyroid and hypothyroid rats, while phentolamine was effective only in hypothyroid rats. In atria from reserpinized rats, the pD2-values for DA in both chronotropic and inotropic effects were reduced, but effectiveness of propranolol or phentolamine on DA-induced positive chronotropic and inotropic effects in euthyroid and hypothyroid rats was similar to that in non-reserpinized rats. In hypothyroid rats, DA increased the maximal rate of tension development, which was inhibited by both phentolamine and propranolol. DA shortened the duration of contraction. DA in the presence of phentolamine significantly shortened the duration of contraction but did not in the presence of propranolol. In conclusion the DA-induced positive chronotropic effect is mainly produced by beta-adrenoceptor stimulation in both euthyroid and hypothyroid rats, and also by a alpha-adrenoceptors to some extent in hypothyroid rats. The DA-induced positive inotropic effect is produced by alpha- as well as beta-adrenoceptor stimulation in both groups. However, alpha-adrenoceptors were involved to a greater extent in hypothyroid rats than in euthyroid rats. The stimulation of alpha-adrenoceptors by DA causes an increase in the maximal rate of tension development without a significant change in the duration of contraction, and the stimulation of beta-adrenoceptors by DA causes an increase in the maximal rate of tension development and shortening of the duration of contraction.     

Influence of lidocaine on ouabain-induced inotropic response in rat atria

In this paper we demonstrated that lidocaine broadens the therapeutic range of ouabain action having a protective effect on ouabain-induced toxicity on rat atria. The lidocaine effect on therapeutic ouabain action was associated with the increase in the sensitivity of Na(+)-K(+)-ATPase related to a decreased in the equilibrium dissociation constant (K(d)) of high affinity binding sites. Lidocaine suppressed the ouabain-induced tonotropic effect and arrhythmias, decreasing the number of low affinity binding sites (B(max)) without changes in K(d). Blockade of Na(+)-Ca(2+) exchange with KB-R7943 or dual Na(+)-Ca(2+) channel with flunarizine, mimicked lidocaine effect increasing ouabain therapeutic action, extending its concentration range tolerated, delaying the onset of contracture. Lidocaine itself triggered negative inotropic response at high concentration. This effect was increased in the presence of flunarizine and verapamil but not by the inhibition of calcium/calmodulin with W-7. The mechanism underlying the lidocaine-induced negative inotropic response, appears to be different that underlying the positive inotropic effect on ouabain action. This study provides evidence that lidocaine can interact with the same or similar binding sites for ouabain in rat atrial tissue, providing a protective effect on ouabain-induced changes in contractility. The contribution of Na(+)-Ca(2+) exchange and/or Ca(2+) overload on lidocaine effect is discussed.     

Age-dependent differences in the positive inotropic effect of phenylephrine on rat isolated atria

The age-dependent differences in the involvement of alpha-adrenoceptors in the positive inotropic effect of phenylephrine (Phe) were examined in isolated atria of male Wistar rats 6 weeks (6W), 10 weeks and 7 months (7M) of age. The maximal increase in tension development induced by Phe increased with age, whereas the EC50 values for the positive inotropic effect of Phe did not change with age. The inhibitory effect of phentolamine on the response to Phe increased with age. Propranolol caused only slight inhibition of the effect of Phe in both 6W and 7M rats, and the EC50 values for Phe in the presence of propranolol did not change significantly with age. The EC50 values for isoprenaline and 5-hydroxytryptamine in 7M rats were higher than those in 6W rats. In 7M rats, the duration of the tension development was only slightly affected by Phe in the presence or absence of propranolol, but it was markedly decreased by Phe in the presence of phentolamine. The dose-response curve for Phe was markedly shifted to the left by papaverine in 6W rats, but slightly in 7M rats. The dose-response curve for isoprenaline was markedly shifted to the left by papaverine in both groups. These results are consistent with effects of Phe being mediated by both alpha- and beta-adrenoceptors in both 6W and 7M rats, but there is a shift in the balance from rather more beta-receptors in the young animals to more alpha-receptors in the adults.     

Modification of the inotropic effect of digoxin by diazepam in rat left atria

There is a pharmacokinetic interaction between digoxin and diazepam that increases the elimination half-life of digoxin. It may be due to a reduction of digoxin tissue concentrations and to an enhanced effect of diazepam on digoxin binding to plasma albumin. Diazepam (10(-5) M) also induces a positive inotropic effect in guinea-pig isolated atria. In a study of a possible pharmacodynamic interaction between both drugs, the inotropic response to digoxin has been examined in rat isolated atria in the presence of diazepam. The atria were kept in Tyrode at 37 degrees C, bubbled with 95% O2 and 5% CO2 and electrically stimulated at twice the threshold voltage. The results indicate that diazepam induces an inotropic effect at 10(-5) M (P less than 0.05) and reduces (P less than 0.05) at 10(-9), 10(-7) and 10(-5) M the inotropic response to digoxin (10(-5) M).     

Safety of dietary supplements: chronotropic and inotropic effects on isolated rat atria

We investigated the effects of dietary supplements on atria isolated from male Wistar rats. The examined supplements, which are increasingly used in Japan, those were Ginkgo biloba extract (GBE), catechins, isoflavones, sodium iron chlorophyllin and sodium copper chlorophyllin. GBE at 100-1000 microg/ml significantly increased the beat rate and the contractile force. Catechins at 1-100 microg/ml significantly potentiated the contractile force but did not effect the beat rates. However, isoflavones, sodium iron and sodium copper chlorophyllins did not change the contractile force or the beat rates. To identify the active ingredient of GBE, ginkgolide B, quercetin and amentoflavone on the atria were tested. Ginkgolide B weakened the contractile force. Quercetin potentiated the contractile force at only 30 microg/ml. Amentoflavone significantly increased the beat rate. From these findings, amentoflavone and quercetin were considered to be the principal ingredients of GBE producing the positive chronotropic and inotropic actions, respectively. In the case of catechins, (-)-epigallocatechin gallate (EGCg), one of the principal ingredients, produced inotropic actions. These findings suggest that there are some dietary supplements which affect cardiac function, such GBE and catechins.     

Mechanisms of the inotropic actions of MPTP and MPP+ on isolated atria of rat

Besides having toxic actions, MPTP (1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine) and MPP+ (1-methyl-4-phenyl-pyridinium ion) produce the release of catecholamines in the peripheral and central nervous systems. This paper reports on the effects of MPTP and MPP+ on isolated left atria of rats and their mechanisms. MPP+ and MPTP produced a concentration-dependent positive inotropic effect. This action was blocked by propranolol and nomifensine; however, inhibition of monoamine oxidase had no effect on the response. In atria from reserpinized rats, the positive inotropic effect of MPTP was also blocked and a negative inotropic effect was unmasked which continued to increase in magnitude during wash. The negative inotropic effect was markedly reduced by superoxide dismutase and catalase. These results indicate that MPTP and MPP+ produce a catecholamine-mediated positive inotropic effect that is not MAO-dependent, unlike the toxic actions of MPTP. These results also suggest that MPTP may directly damage cardiac muscle by generating free radicals which might explain why high doses of MPTP are lethal to animals.     

The mechanism of the positive inotropic action of ketamine on isolated atria of the rat.

1 The effect of ketamine (10-7 M-5 X10-4 M) on electrical and mechanical properties of isolated atria of the rat was investigated. 2. On spontaneously beating right atria, ketamine produced a dose-dependent positive inotropic effect which was accompanied by a progressive reduction in atrial rate. 3 In electrically driven left atria, ketamine produced a dose-dependent positive inotropic effect which was accompanied by a parallel increase in df/dtmax and by a prolongation in the time to peak tension and in the time for total concentration. The positive inotropic effect occurred concomitantly with an increase in the height and duration of the plateau phase of the action potential of atrial fibres. 4 The positive inotropic effect of ketamine varied with the concentration of Ca and Na in the bathing media and the rate of stimulation. 5 Ketamine decreased post-extrasystolic potentiation and the amplitude-interval relationship. 6 The positive inotropic effect of ketamine was inhibited by verapamil (10-6 M) and by caffeine (4 X10-3 M). 7 Ketamine, 5 X10-5M and 10-4M, increased 45Ca uptake in electrically driven left atria. At 10-4M and 5 X10-4M, ketamine also increased 45Ca efflux. 8 These results suggest that ketamine produces its positive inotropic effect by two possible mechanisms. One of these is presumed to be an effect on the cell membrane with leads to an increased Ca influx into atrial fibres; the other is probably related to the inhibition of Ca sequestration by the sarcoplasmic reticulum.     

Lymphocytes sensitize rat isolated atria to the inotropic and chronotropic effects of sodium arachidonate

Normal human lymphocytes (4 X 10(5) ml-1) incubated with sodium arachidonate (8 X 10(-7)M) (NaA-L) induced a strong enhancement of the tension and frequency of spontaneously beating rat atria. Normal human lymphocytes (L) or NaA alone at 8 X 10(-7)M did not modify this contractile activity. Between 2 X 10(-6)M to 1 X 10(-5)M NaA alone increased the tension of the atria without effect on the rate. In the presence of L (4 X 10(5) ml-1) the dose-response curve to NaA shifted to the left, the potency and the efficiency of NaA were enhanced and the chronotropic action was triggered. Inhibitors of cyclo-oxygenase (indomethacin 1 X 10(-6)M or acetylsalicylic acid (ASA) 1.8 X 10(-4)M) completely blocked the positive inotropic effect induced by NaA alone. Inhibitors of lipoxygenase/s (nordihydroguaiaretic acid (NDGA) 1 X 10(-5)M or 5,8,11,14-eicosatetraynoic acid (ETYA) 1 X 10(-7)M did not modify this effect. Indomethacin and ASA did not block the positive inotropic and chronotropic effects of the lower concentration of NaA-L and significantly reduced the inotropic effect of the higher ones. NDGA and ETYA shifted to the right the inotropic and chronotropic dose-response curve to NaA-L. FPL-55712 (1 X 10(-7)M), the slow reacting substance of anaphylaxis (SRS-A) antagonist, significantly reduced the overall inotropic and chronotropic effect of NaA-L. Direct contact of NaA-L with the atria was not necessary. Cell-free supernatants of L exposed to NaA increased the tension and the frequency of beating rat atria.(ABSTRACT TRUNCATED AT 250 WORDS)     

Cardiotoxic and inotropic effects of ouabain on atria isolated from rabbits of different ages.

1 In sino-atrial node-right atrial preparations and left atrial preparations obtained from rabbits of different ages (2 to 360 days old), cardiotoxic and inotropic effects of ouabain were compared. Tachyarrhythmias were produced in immature and mature rabbit atria exposed for 6 to 8 min to ouabain (5 x 109(-6) M). The total number of contractions before the arrhythmias developed varied inversely with age (2 to 90 days old). Percentage reductions in the resting membrane potential, overshoot and maximum rate of rise induced by ouabain were greater in mature rabbit atria than in immature rabbit atria. 2 Reduction of Ca2+ in the bathing media suppressed the development of tachyarrhythmias. 3 In electrically-driven left atria isolated from immature and mature rabbits, similar frequency-contractile force curves were obtained, maximum contractions being attained at a frequency of 120/min. Ouabain at 2 x 10(-7) M shifted the curve upward and at 10(-6) M greatly potentiated the contractile force developed at low frequencies (6 to 30/min). Increase in contractions induced by ouabain relative to pre-drug concentrations at a frequency of 60/min were greater in immature rabbit atria (15 days old) but the positive inotropic effect of 2 x 10(-7) M ouabain relative to the effect seen at 10(-6) M was appreciably less. 4 It may be concluded that atria isolated from immature rabbits tolerate higher concentrations of ouabain than those isolated from mature rabbits.     

Characterization of the 5-hydroxytryptamine receptor mediating the positive inotropic response in guinea-pig isolated left atria.

1. 5-Hydroxytryptamine (5-HT), in the presence of propranolol (1 microM), atropine (3 microM) and ketanserin (1 microM), induced a positive inotropic response of guinea-pig isolated electrically paced left atria (pEC50 = 7.52). The positive inotropic response was mimicked by alpha-methyl-5-HT (pEC50 = 7.26) and 5-carboxamidotryptamine (5-CT; pEC50 = 6.56) but not by sumatriptan or 1-(m-chlorophenyl) piperazine (m-CPP). 2. The 5-HT induced positive inotropic response was competitively antagonized by both mesulergine (pA2 = 7.68) and methiothepin (pA2 = 6.67). Methysergide was a surmountable antagonist at 3 nM producing a rightward shift in the 5-HT concentration-response curve giving an apparent pA2 = 9.2 with no significant reduction in the maximum. At higher concentrations, methysergide behaved as an insurmountable antagonist, significantly reducing the maximum response to 5-HT as well as producing rightward shifts in the 5-HT concentration-response curves. 3. The 5-HT-induced positive inotropic response was not antagonized by either tropisetron (10 microM) or yohimbine (10 microM). 4. The guinea-pig atrial 5-HT receptor does not satisfy the criteria for any of the currently recognised 5-HT receptor subtypes and appears to have some similarities to the atypical 5-HT receptors previously described in other peripheral tissues.     

Influence of thyroid hormone on the positive inotropic effects mediated by alpha- and beta-adrenoceptors in isolated guinea pig atria and rabbit papillary muscles.

Effects of thyroxine treatment for 7--11 days on the positive inotropic effects mediated by alpha- and beta-adrenoceptors were studied in isolated guinea pig atria and rabbit papillary muscles. In guinea pig atria, the thyroxine treatment inhibited the positive inotropic effect of lower concentrations of phenylephrine (PHE), and attenuated the inhibitory effect of phentolamine on the PHE response. The effect of isoproterenol (ISO) was potentiated by the thyroxine treatment. In rabbit papillary muscles, the thyroxine treatment shifted the dose--response curve for PHE to the right and attenuated the inhibitory effect of phentolamine on the PHE response. Propranolol, in both guinea pig atria and rabbit papillary muscles, inhibited the PHE response more effectively in preparations from thyroxine-treated animals than in controls. In guinea peg atria, the attenuation of the PHE response mediated by alpha-adrenoceptors was observed after the thyroxine treatment for only 2 days, whereas the potentiation of the ISO response required the thyroxine treatment for a longer period. It was concluded that the thyroxine treatment attenuated the positive inotropic effect mediated by alpha-adrenoceptors and potentiated that mediated by beta-adrenoceptor-mediated positive inotropic effects due to the thyroxine treatment may be independent of each other.     

甲基葡糖苷对豚鼠离体心房的正性肌力作用

目的　观察甲基葡糖苷对豚鼠离体心房的正性肌力作用 ,并探讨其作用机制。方法　采用豚鼠离体左右心房 ,测定药物对心房肌收缩力、右心房心率 ,以及对静息后收缩和正阶梯现象的影响 ,并测定大鼠心肌细胞膜Na+ K+ ATP酶活性。结果　甲基葡糖苷显著增强心房肌收缩力 ,减慢右心房心率 ,且呈剂量依赖性 ;能明显增强左心房静息后收缩和正阶梯现象 ,并能显著抑制大鼠心肌细胞膜Na+ K+ ATP酶活性。结论　甲基葡糖苷具有加强心房肌收缩力 ,降低右心房心率的作用 ,其正性变力作用可能与抑制心肌细胞膜Na+ K+ ATP酶活性 ,促进心肌细胞外钙内流和内钙释放有关     

Comparison of the inotropic effects of some 5 alpha-cardenolides on guinea pig left atria

The inotropic activity of eleven 5 alpha-cardenolides and six 5 beta-cardenolides was determined using guinea pig left atria and the results were compared with published data obtained from cat toxicity studies. The guinea pig atrial studies showed that the configuration of the A/B ring junction did not influence significantly the cardiotonic potency of digitoxigenin and its 5 alpha-epimer, uzarigenin, but was of significance with respect to the influence of substituent groups. Glucosidation decreased the potency of uzarigenin but increased that of digitoxigenin. By contrast, conjugation with rhamnose increased the activity of both uzarigenin and digitoxigenin. Cat toxicity data did not correlate well with that obtained using guinea pig atria, possibly because of the variable influence of pharmacokinetic factors applicable to the cat toxicity studies.     

Modulatory effects of diltiazem on inotropic responses to amrinone on rabbit isolated atria

The effect of pretreatment with graded concentration of diltiazem on the inotropic responses to amrinone were studied on isolated atria of rabbit. The responses to amrinone were modified by diltiazem in a biphasic manner; initial potentiation followed by inhibition. The potentiation is proposed to be due to synergistic rise in cytosolic calcium ion concentration by diltiazem and amrinone. The inhibition by diltiazem in higher concentration may be due to blockade of calcium ion influx and depletion of intracellular calcium ion from storage sites.