Negative Inotropic Effect Of Adenosine Research Articles

Adrenergic and adenosinergic regulation of the cardiovascular system in an Antarctic icefish: Insight into central and peripheral determinants of cardiac output

Icefishes characteristically lack the oxygen-binding protein haemoglobin and therefore are especially reliant on cardiovascular regulation to augment oxygen transport when oxygen demand increases, such as during activity and warming. Using both in vivo and in vitro experiments, we evaluated the roles for adrenaline and adenosine, two well-established cardio- and vasoactive molecules, in regulating the cardiovascular system of the blackfin icefish, Chaenocephalus aceratus. Despite increasing cardiac contractility (increasing twitch force and contraction kinetics in isometric myocardial strip preparations) and accelerating heart rate (ƒH), adrenaline (5 nmol kg−1 bolus intra-arterial injection) did not significantly increase cardiac output (Q̇) in vivo because it elicited a large decrease in vascular conductance (Gsys). In contrast, and despite preliminary data suggesting a direct negative inotropic effect of adenosine on isolated atria and little effect on isolated ventricle strips, adenosine (500 nmol kg−1) generated a large increase in Q̇ by increasing Gsys, a change reminiscent of that previously reported during both acute warming and invoked activity. Our data thus illustrate how Q̇ in C. aceratus may be much more dependent on peripheral control of vasomotor tone than direct regulation of the heart.

FSCPX, a Chemical Widely Used as an Irreversible A₁ Adenosine Receptor Antagonist, Modifies the Effect of NBTI, a Nucleoside Transport Inhibitor, by Reducing the Interstitial Adenosine Level in the Guinea Pig Atrium.

Based on in silico results, recently we have assumed that FSCPX, an irreversible A1 adenosine receptor antagonist, inhibits the action of NBTI that is apparent on E/c curves of adenosine receptor agonists. As a mechanism for this unexpected effect, we hypothesized that FSCPX might modify the equilibrative and NBTI-sensitive nucleoside transporter (ENT1) in a way that allows ENT1 to transport adenosine but impedes NBTI to inhibit this transport. This assumption implies that our method developed to estimate receptor reserve for agonists with short half-life such as adenosine, in its original form, overestimates the receptor reserve. In this study, therefore, our goals were to experimentally test our assumption on this effect of FSCPX, to improve our receptor reserve-estimating method and then to compare the original and improved forms of this method. Thus, we improved our method and assessed the receptor reserve for the direct negative inotropic effect of adenosine with both forms of this method in guinea pig atria. We have found that FSCPX inhibits the effects of NBTI that are mediated by increasing the interstitial concentration of adenosine of endogenous (but not exogenous) origin. As a mechanism for this action of FSCPX, inhibition of enzymes participating in the interstitial adenosine production can be hypothesized, while modification of ENT1 can be excluded. Furthermore, we have shown that, in comparison with the improved form, the original version of our method overestimates receptor reserve but only to a small extent. Nevertheless, use of the improved form is recommended in the future.

Approximation of A1 adenosine receptor reserve appertaining to the direct negative inotropic effect of adenosine in hyperthyroid guinea pig left atria

Hyperthyroidism elevates cardiovascular mortality by several mechanisms, including increased risk of ischemic heart disease. Therefore, therapeutic strategies, which enhance tolerance of heart to ischemia-reperfusion injury, may be particularly useful for hyperthyroid patients. One promising cardioprotective approach is use of agents that cause (directly or indirectly) A1 adenosine receptor (A1 receptor) activation, since A1 adenosinergic pathways initiate protective mechanisms such as ischemic preconditioning. However, previously we found great A1 receptor reserve for the direct negative inotropic effect of adenosine in isolated guinea pig atria. This phenomenon suggests that weakening of atria is a possible side effect of A1 adenosinergic stimulant agents. Thus, the goal of the present investigation was to explore this receptor reserve in hyperthyroidism. Our recently developed method was used that prevents the rapid intracellular elimination of adenosine, allowing sufficient time for exogenous adenosine administered for the generation of concentration-response curves to exert its effect. Our method also allowed correction for the bias caused by the consequent endogenous adenosine accumulation. Our results demonstrate that thyroxine treatment does not substantially affect the A1 receptor reserve for the direct negative inotropic effect of adenosine. Consequently, if an agent causing A1 receptor activation is administered for any indication, the most probable adverse effect affecting the heart may be a decrease of atrial contractility in both eu- and hyperthyroid conditions.

The guinea pig atrial A1 adenosine receptor reserve for the direct negative inotropic effect of adenosine

Although the A1 adenosine receptor (A1 receptor), the main adenosine receptor type in cardiac muscle, is involved in powerful cardioprotective processes such as ischemic preconditioning, the atrial A1 receptor reserve has not yet been quantified for the direct negative inotropic effect of adenosine. In the present study, adenosine concentration-effect (E/c) curves were constructed before and after pretreatment with FSCPX (8-cyclopentyl-N3-[3-(4-(fluorosulfonyl)benzoyloxy)propyl]-N1-propylxanthine), an irreversible A1 receptor antagonist, in isolated guinea pig atria. To prevent the intracellular elimination of the administered adenosine, NBTI (S-(2-hydroxy-5-nitrobenzyl)-6-thioinosine), a nucleoside transport inhibitor, was used. As expected, NBTI alone and FSCPX-pretreatment alone shifted the adenosine E/c curve to the left and right, respectively. However, in the presence of NBTI, FSCPX-pretreatment appeared to increase the maximal response to adenosine. By means of the receptorial responsiveness method (RRM), our recently developed procedure, adenosine E/c curves generated in the presence of NBTI were corrected for the bias caused by the endogenous adenosine accumulated by NBTI. The corrected curves indicate a substantial A1 receptor reserve for the direct negative inotropy evoked by adenosine. In addition, our results suggest that accumulation of an endogenous agonist may bias the E/c curve constructed with the same or similar agonist that can lead to seemingly paradoxical results.

Thyroid hormones decrease the affinity of 8-cyclopentyl-1,3-dipropylxanthine (CPX), a competitive antagonist, for the guinea pig atrial A1 adenosine receptor

The aim of the present study was to investigate whether or not thyroxine (T(4)) treatment affects K(B), the equilibrium dissociation constant of the antagonist-receptor complex, for the interaction between CPX, a selective and competitive orthosteric antagonist, and the guinea pig atrial A1 adenosine receptor A1 receptor). The inotropic response to adenosine, a nonselective adenosine receptor agonist, or CPA, a selective A1 receptor agonist, was investigated in the absence or presence of CPX in paced left atria isolated from 8-day solvent- or T(4)-treated guinea pigs. To obtain K(B) values, adenosine and CPA concentration-response curves were evaluated by Schild analysis. CPA but not adenosine obeyed the requirements of the Schild analysis to provide correct K(B) values for CPX. According to the CPA concentration-response curves, affinity of CPX for the hyperthyroid guinea pig atrial A1 receptor (K(B) = 44.16 nM) was lower than that for the euthyroid one (K(B) = 16.63 nM). Regarding the intense reduction in the negative inotropic effect of adenosine and CPA in hyperthyroid atria, it is reasonable to assume that the moderate decrease in affinity of the guinea pig atrial A1 receptor is only in part responsible for the diminished A1 receptor-mediated effect in hyperthyroidism.

Adenosine induced direct negative inotropic effect is abolished during global ischemia: role of protein kinase C and prostacyclin

Adenosine acts as a cardioprotective agent by producing coronary vasodilation, decreasing heart rate and by antagonizing the cardiostimulatory effect of catecholamines; adenosine also exerts a direct negative inotropic effect. Myocardial ischemia is known to be associated with enhanced levels of adenosine, increased protein kinase C (PKC) activity and prostacyclin (PGI2) release. The present study was conducted to determine if myocardial ischemia alters the cardioprotective effect of adenosine by increasing PKC activity and PGI2release in the isolated rat heart perfused at 10 ml/min with Krebs–Henseleit buffer (KHB; 95% O2+5% CO2). Adenosine (10 mmol/min) reduced myocardial contractility as indicated by a decrease in contractility (dp/dtmax), heart rate (HR) and coronary perfusion pressure (PP). In hearts subjected to 30 min of ischemia (without perfusion) and then reperfused with KHB, adenosine failed to decrease dp/dtmax, HR or PP. However, during infusion of PKC inhibitor H-7 (1-(5-Isoquinolinesulfonyl)-2-methylpiperazine hydrochloride) (H-7; 6 mmol/min), which commenced 10 min before ischemia and continued throughout reperfusion, adenosine produced a decrease in dp/dtmax, HR and PP, similar to that before ischemia. Infusion of the PKC activator phorbol 12,13-dibutyrate (PDBu; 2 nmol/min) but not an inactive analogue in non-ischemic hearts prevented the adenosine induced decrease in dp/dtmax. During infusion of H-7, PDBu failed to block the direct negative inotropic effect of adenosine in non-ischemic hearts. In addition, pretreatment with H-7 or indomethacin (cyclooxygenase inhibitor) significantly reduced the PGI2release following ischemia. This data suggest that PKC and PGI2regulate the direct negative inotropic effect of adenosine, which is abolished during ischemia.

FK453: A Novel Non‐Xanthine Adenosine A1 Receptor Antagonist as Diuretic

Adenosine is a purine nucleotide that is widely distributed throughout the body; it modulates a wide variety of physiological functions via extracellular receptors. Adenosine receptors, located on the cell membranes, are classified into two major subtypes, adenosine A, and A, receptors. This classification was originally based on the effects of a series of adenosine analogs on adenylate cyclase activity; activation of the adenosine A, receptor inhibits, while activation of the adenosine A2 receptor stimulates, adenylate cyclase (26,46). Many adenosine agonists that have a preferential affinity for adenosine A, receptors depress heart rate, myocardial contractility. and impulse conduction velocity (5,10,14). The agonists that have a greater affinity for adenosine A, receptors induce vasodilation ( 1 1.15,17,24,28). A number of workers have found that another adenosine receptor, which has been classified as an adenosine A , receptor, is present in many tissues. This receptor was recently subdivided into adenosine A, and A, receptor subtypes. As knowledge of the different types of adenosine receptors increases, these receptors may prove to be possible targets for novel pharmacologically active drugs. Adenosine itself is used in the acute treatment and diagnosis of supraventricular arrhythmias. To date, however, no selective adenosine receptor agonist or antagonist has been used therapeutically. FK453 is a novel pyrazolopyridine derivative that was synthesized at Fujisawa Pharmaceutical Co., Ltd. (Osaka, Japan). During a search for diuretics, FK453 was found to possess a potent diuretic activity with renal vasodilatory and uricosuric effects (40). In addition. FK453 markedly antagonized the negative inotropic effects of adenosine in isolated guinea pig atria, but was less potent in inhibiting the relaxation induced by adenosine in guinea pig aorta. These results suggested that FK453 is a potent and selective

Increased responses to adenosine in isolated left atria from streptozotocin-diabetic rats: evidence for the involvement of hypothyroidism.

Direct and indirect (antiadrenergic) inotropic effects of adenosine in the isolated left atrial preparations from streptozotocin-induced diabetic rats were examined by comparing with those from propylthiouracil-induced hypothyroid rats. Experimental diabetes was induced by a single i.v. injection of streptozotocin (45 mg/kg). Subsets of diabetic rats were treated daily with either insulin (6-8 units/kg) or triiodothyronine (T3; 8-10 micrograms/kg). After 10 weeks, negative inotropic and antiadrenergic effects of adenosine were assessed in the atria from nondiabetic, diabetic, insulin- or T3-treated diabetic and hypothyroid rats. Diabetic rats exhibited a significant increase in negative inotropic and antiadrenergic effects of adenosine, as well as signs of hypothyroidism such as decreased thyroid hormone levels. The same changes in the adenosine effects also were noticed in the hypothyroid rats. In our study, the influence of T3 treatment on the hyperreactivity of diabetic rat atria to adenosine also was examined by comparing with that of insulin treatment. Both insulin and T3 treatments restored the hyperreactivity to adenosine, with the exception of adenosine receptor affinity (pD2 value) calculated for the negative inotropic effect of adenosine. These findings suggest that thyroid hormone deficiency is likely to be involved in the inotropic and antiadrenergic effects of adenosine on the left atria from rats with experimental diabetes for 10 weeks.

The Role of Gi-proteins andβ-Adrenoceptors in the Age-related Decline of Contraction in Guinea-pig Ventricular Myocytes

A decline in contractility in myocytes from ageing guinea-pig hearts was demonstrated, which is more pronounced for maximumβ-adrenoceptor-stimulated activity than contraction in high Ca2+. In this study the role of the inhibitory G-proteins (Gi) in this process was investigated. Comparisons were made between young (Y, <400 g, <4 weeks), adult (A, >600 g, >8 weeks) and senescent guinea pigs (S, 53–65 weeks, 1136±30 g). Giαactivity, detected by pertussis toxin-catalysed ADP ribosylation, was significantly increased in senescent compared to young animals, but immunodetectable levels of Giαwere unchanged.β-adrenoceptor number was decreased by 27% in senescent compared with young animals (P<0.002). Pertussis toxin treatment increased the maximum response to isoproterenol in contacting myocytes so that there was no longer any significant decline with age. Maximum contraction amplitudes (sarcomere length change,μm) with isoproterenol before pertussis toxin were 0.144±0.011 (Y,n=22 animals), 0.104±0.009 (A, 18) and 0.098±0.009 (S, 14),P<0.01 by analysis of variance (ANOVA). Following toxin treatment amplitudes were 0.140±0.012 (Y, 12), 0.117±0.010 (A, 10) and 0.117±0.018 (S, 8),P=n.s. Pertussis toxin treatment also reversed the effects of ageing on contraction and relaxation velocity in isoproterenol. In contrast, the effect of age on contraction amplitude or velocity in maximum Ca2+was more pronounced after toxin treatment. The EC50value for isoproterenol increased with age: pertussis treatment decreased the EC50in each group, but the effect was especially pronounced for senescent animals. There was no significant difference in the concentration–response curves for the negative inotropic effect of adenosine (in the presence of isoprotenerol) between the three age groups before toxin treatment. All effects of adenosine were abolished after pertussis exposure. We conclude that increased Giαactivity is likely to contribute to the decreased response to isoproterenol, but not to high Ca2+, in myocytes from ageing guinea-pigs.

Supersensitivity mismatch of adenosine in the transplanted human heart: chrono- and dromotropy versus inotropy

Abstract Supersensitive negative chronotropic and dromotropic effects have been described for adenosine after human heart transplantation. The present study investigated a potential antiadrenergic negative inotropic effect of adenosine in heart transplant recipients compared to normal subjects. Sinus cycle length, PR interval, blood pressure, and inotropic response in vivo were compared in seven ortho-topic heart transplant recipients and seven healthy volunteers (controls). Fractional shortening, velocity of circumferential fiber shortening, and systolic pressure/dimension ratio were calculated using M-mode echocardiography. Baseline ventricular contractility was normal in both groups. Although adenosine induced a significant exaggeration of the negative chronotropic and dromotropic effect in the transplant group, the positive inotropic effect of 20 ng/kg × min isoproterenol (FS 53.2 ± 8.8 vs 51.0 ± 4.6 %, P/D 5.8 ± 1.9 vs 6.0 k 0.8 mm Hg/mm, Vcf 0.21 ± 0.04 vs 0.20 ± 0.02 %/ms for heart recipients vs controls) was not reduced by the additional administration of 150 pg/kg adenosine (FS 52.2 ± 8.6 vs 51.7 ± 5.6 %, P/D 5.5 ± 1.5 vs 5.4 ± 0.8 mm Hg/mm, Vcf 0.24 ± 0.07 vs 0.21 k 0.02 %/ms for transplant recipients vs controls). In contrast to a chronotropic and dromotropic supersensitivity, adenosine does not attenuate the catecholamine-induced increase in contractility in the human ventricle in vivo after heart transplantation.

Supersensitivity mismatch of adenosine in the transplanted human heart: chrono- and dromotropy versus inotropy

Supersensitive negative chronotropic and dromotropic effects have been described for adenosine after human heart transplantation. The present study investigated a potential antiadrenergic negative inotropic effect of adenosine in heart transplant recipients compared to normal subjects. Sinus cycle length, PR interval, blood pressure, and inotropic response in vivo were compared in seven orthotopic heart transplant recipients and seven healthy volunteers (controls). Fractional shortening, velocity of circumferential fiber shortening, and systolic pressure/dimension ratio were calculated using M-mode echocardiography. Baseline ventricular contractility was normal in both groups. Although adenosine induced a significant exaggeration of the negative chronotropic and dromotropic effect in the transplant group, the positive inotropic effect of 20 ng/kg x min isoproterenol (FS 53.2 +/- 8.8 vs 51.0 +/- 4.6%, P/D 5.8 +/- 1.9 vs 6.0 +/- 0.8 mm Hg/mm, Vcf 0.21 +/- 0.04 vs 0.20 +/- 0.02%/ms for heart recipients vs controls) was not reduced by the additional administration of 150 micrograms/kg adenosine (FS 52.2 +/- 8.6 vs 51.7 +/- 5.6%, P/D 5.5 +/- 1.5 vs 5.4 +/- 0.8 mm Hg/mm, Vcf 0.24 +/- 0.07 vs 0.21 +/- 0.02%/ms for transplant recipients vs controls). In contrast to a chronotropic and dromotropic supersensitivity, adenosine does not attenuate the catecholamine-induced increase in contractility in the human ventricle in vivo after heart transplantation.

Loss of adenosine-induced negative inotropic effect in hyperexcited rabbit hearts: relationship to PKC

Adenosine produced a negative inotropic effect in hearts isolated from calm rabbits but not from those exhibiting alarm behavior during handling. This study was conducted to determine whether protein kinase C (PKC) activation is responsible for the loss of adenosine-induced negative inotropism in the hearts of hyperexcited rabbits. Adenosine (10 microM) decreased myocardial contractility (dP/dtmax) in the hearts of calm, but not hyperexcited, rabbits but decreased heart rate (HR) and coronary perfusion pressure (PP) in the hearts of both calm and hyperexcited animals. During infusion of calphostin C (200 nM), a PKC inhibitor, adenosine also decreased dP/dtmax in the hearts of hyperexcited rabbits. Calphostin C did not alter the actions of adenosine in the hearts of calm rabbits. Agents that stimulate PKC directly [phorbol 12,13-dibutyrate (PDBu), 1 nM] or indirectly [norepinephrine (NE), 3 nM; angiotensin II (ANG II), 5 nM] abolished the adenosine-induced decrease in dP/dtmax but not HR or PP in the hearts of calm rabbits. During calphostin C, infusion of PDBu, NE, and ANG II failed to prevent the adenosine-induced decrease in dP/dtmax. These data suggest that the lack of a negative inotropic effect of adenosine in hyperexcited rabbits is due to an increase in PKC activity.

Mechanism of the negative inotropic effect of adenosine in guinea pig atrial myocytes.

The ionic basis of the negative inotropic effect of adenosine on guinea pig atrial myocytes was studied. Membrane potentials and currents were measured using a whole cell patch-clamp technique. The contractility was assessed by video quantitation of cell twitch amplitude. Adenosine shortened action potential duration [measured at 90% repolarization (APD90)] and decreased twitch amplitude in a concentration-dependent manner. The maximal effects of adenosine (100 microM) were to reduce APD90 from 102 +/- 14 to 34 +/- 8 ms (n = 11) and twitch amplitude from 4.3 +/- 0.9 to 1.5 +/- 0.4 microns (n = 8). The concentration of adenosine that caused one-half of the maximal reductions of twitch amplitude and of APD90 was 0.6 microM. Reductions in APD90 and in twitch amplitude were parallel and highly correlated (r = 0.98). Decreases in twitch amplitude by adenosine could be mimicked by application of voltage-clamp pulses with durations similar to the durations of action potentials in the presence of adenosine. Clamp pulse could reverse adenosine-induced but not cadmium chloride-induced decreases in twitch amplitude. Adenosine activated the inwardly rectifying K+ current (IK,Ado), but did not significantly decrease the L-type Ca2+ current (ICa,L). Adenosine reduced the effects of BAY K 8644 on APD90 and twitch amplitude but did not attenuate the BAY K-induced increase in ICa,L. The effects of adenosine on APD90 and twitch amplitude could be reversed after activation of IK,Ado was inhibited by intracellular application of cesium and tetraethylammonium chloride.(ABSTRACT TRUNCATED AT 250 WORDS)

Pertussis toxin does not affect the adenosine-induced inhibition of the efferent function of cardiac capsaicin-sensitive nerves.

The negative inotropic effect of adenosine (1-100 microM) was abolished in isolated guinea-pig atria obtained from pertussis toxin-pretreated guinea pigs electrically driven at 4 Hz. However, the inhibitory effect of the same concentrations of adenosine on the cardiac response to stimulation of non-adrenergic non-cholinergic (NANC), capsaicin-sensitive sensory nerves, was not modified by the toxin. These results suggest that, while pertussis toxin-sensitive G proteins are involved in the negative inotropic effect of adenosine, they do not mediate the inhibitory effect of adenosine on cardiac NANC neurotransmission.

Vasodilative and anti-adrenergic effects of adenosine in diabetic rat hearts.

To determine the vasodilative and negative inotropic effects of adenosine in hearts of diabetic rats, isolated hearts, perfused at constant perfusion pressure (Langendorff technique), were prepared from age-matched control Wistar rats and rats made diabetic 10 weeks prior to study by a single injection of streptozotocin (65 mg.kg-1, i.p.). Adenosine and nitroprusside each increased coronary inflow when administered either as bolus injections or as infusions. Coronary flow responses to nitroprusside were unchanged in diabetic hearts. Coronary flow responses of diabetic hearts to adenosine injections were unchanged, but responses to adenosine infusions tended to be larger than in normal hearts. Diabetes had no significant effect on the EC50 for either vasodilator. Adenosine inhibited the inotropic effect of isoproterenol (enhanced left ventricular (LV) pressure (P) and LV dP/dtmax) in normal hearts, independently of its vasodilative action. This negative inotropic action of adenosine appeared equally strong in diabetic hearts. We conclude that adenosine's coronary vasodilative and anti-beta-adrenergic, negative inotropic effects in the rat heart were not diminished after 10 weeks of streptozotocin-induced diabetes mellitus. Thus, earlier reports of diminished adenosine dilative efficacy in experimental diabetes may have been unique to those particular models.

Pharmacological characteristics of adenosine-induced inhibition of dog ventricular contractility: dependence on the pre-existing level of ?-adrenoceptor activation

Experiments were carried out to characterize the adenosine-induced negative inotropic effect in relation to the extent of beta-adrenoceptor activation in the isolated dog left ventricular myocardium. Adenosine and R-N6-phenylisopropyladenosine inhibited the positive inotropic effect of isoprenaline (10(-7) mol/l and lower) about 20% of its maximal response, which was antagonized by an A1 adenosine receptor antagonist 1,3-dipropyl-8-cyclopentylxanthine in a concentration-dependent manner. The negative inotropic effect of adenosine disappeared and that of R-N6-phenylisopropyl-adenosine decreased when the isoprenaline concentration was elevated to the level higher than 10(-7) mol/l. Adenosine deaminase (1.5 U/ml) that abolished the negative inotropic effect of adenosine enhanced the effect of R-N6-phenylisopropyladenosine, indicating that endogenous adenosine released by high isoprenaline concentration (10(-6) mol/l) modulates the interaction. The maximal response to adenosine and R-N6-phenylisopropyladenosine determined in the presence of 10(-7) mol/l isoprenaline was 50% of that of carbachol which elicited the maximal inhibition even in the presence of 10(-6) mol/l isoprenaline. The negative inotropic effects of R-N6-phenylisopropyladenosine and carbachol were additive to the maximal response equivalent to that of carbachol. The difference in the efficiency between the adenosine and muscarinic receptor agonists may be partly ascribed to the difference in densities of the respective receptors in the dog ventricular myocardium. The negative inotropic effect of R-N6-phenylisopropyladenosine in the presence of isoprenaline was associated with decrease in cyclic AMP levels elevated previously by isoprenaline.(ABSTRACT TRUNCATED AT 250 WORDS)

Activation of Na+/Ca2+ exchange by adenosine in ewe heart sarcolemma is mediated by a pertussis toxin-sensitive G protein.

We studied the effect of adenosine on Na+/Ca2+ exchange activity in ewe heart ventricular sarcolemmal vesicles. Adenosine was found to stimulate Na+/Ca2+ exchange activity in a dose-dependent manner from 0.1 nM to 10 microM, with maximal stimulation (40%) at 0.1 microM adenosine. The Vmax of Na+/Ca2+ exchange was increased, but the Km for Ca2+ was not altered. The effect of adenosine was specific since 1 microM adenine, inosine, and guanosine led to less than 15% stimulation, and adenosine diphosphate had no effect. Caffeine antagonized the activation of Na+/Ca2+ exchange by adenosine, and the order of potency of adenosine analogs was N6-(L-2-phenylisopropyl)adenosine = N6-cyclohexyladenosine = 5'-(N- ethylcarboxamido)adenosine much greater than N6-(D-2-phenylisopropyl)adenosine, indicating the involvement of A1 subclass receptors. The effect of adenosine was mimicked by guanosine 5'-O-(3-thiotriphosphate) (GTP gamma S) and blocked by pertussis toxin treatment. Taken together, these results suggest that A1 subclass receptors coupled to a pertussis toxin-sensitive G protein mediate the activation of Na+/Ca2+ exchange activity by adenosine. We conclude that the negative inotropic effect of adenosine in ventricular muscle, antagonistic toward cyclic AMP, may involve activation of Na+/Ca2+ exchange.

Characterization of adenosine receptors in guinea-pig isolated left atria.

1. The effects of purinergic stimulation on action potential, force of contraction, 86Rb efflux and 45Ca uptake were investigated in guinea-pig left atria. 2. Adenosine exerted a negative inotropic effect which was antagonized by adenosine deaminase but enhanced by dipyridamole. 3. The negative inotropic effect of adenosine was mimicked by 5'-(N-ethyl)-carboxamido-adenosine (NECA) and the isomers of N6-(phenyl-isopropyl)-adenosine, R-PIA and S-PIA. NECA and R-PIA were about 100 times more potent than adenosine, whereas R-PIA was about 100 times more potent than S-PIA. 4. The inotropic effects of adenosine (in the presence of dipyridamole), NECA, R-PIA and S-PIA were competitively antagonized either by theophylline (pA2 about 4.5) or 8-phenyltheophylline (pA2 about 6.3). 5. NECA and R-PIA shortened the action potential duration and increased the rate constant of the efflux of 86Rb in a concentration-dependent manner with no differences in potency; the effects were competitively antagonized by 8-phenyltheophylline. 6. Barium ions reduced the efflux of 86Rb under control conditions and antagonized the increase induced by NECA and R-PIA. 7. NECA and R-PIA significantly reduced 45Ca uptake in beating preparations. 8. It is concluded that adenosine, NECA and R-PIA activate a common receptor population (P1 or A3) on the outside of the cell membrane of atrial heart muscle to increase the potassium conductance and to reduce the action potential and, thereby, calcium influx and force of contraction.

Alinidine antagonizes the myocardial effects of adenosine

The bradycardiac drug alinidine (2.8–182.3 μM) antagonized the negative inotropic effect of adenosine in isolated left rat atria in a non-competitive fashion. A 50% reduction of the maximal adenosine effect was achieved with 64.6 μM alinidine. The shortening of the action potential duration and the suppression of slow action potentials in guinea pig atria by 1–10 μM adenosine could be abolished by 5.7 μM alinidine. Adenosine (1 mM) exerted a negative inotropic effect on isolated strips from the right ventricle of rat heart exposed to 0.36 μM isoproterenol, an effect which was fully antagonized by 91.1 μM alinidine. Addition of 1 mM adenosine to spontaneously beating isolated rat atria induced pronounced bradycardia. Under these conditions, the bradycardiac agent alinidine increased the contractile rate concentration dependently (11.4–182.3 μM), while the chemically unrelated bradycardiac compound UL-FS 49 did not accelerate the beat frequency. As alinidine antagonized all the cardiac actions of adenosine that we tested, it may be concluded that the drug interferes with A 1-receptor-mediated effects.

3H]-8-cyclopentyl-1,3-dipropylxanthine binding to A1 adenosine receptors of intact rat ventricular myocytes.

The purpose of the present study was the identification of A1 adenosine receptors in intact rat ventricular myocytes, which are thought to mediate the negative inotropic effects of adenosine. The adenosine receptor antagonist [3H]-8-cyclopentyl-1,3-dipropylxanthine was used as radioligand. Binding of the radioligand to intact myocytes was rapid, reversible, and saturable with a binding capacity of 40,000 binding sites per cell. The dissociation constant of the radioligand was 0.48 nM. The adenosine receptor antagonists 8-cyclopentyl-1,3-dipropylxanthine, "xanthine amine congener," and theophylline were competitive inhibitors with affinities in agreement with results obtained for A1 receptors in other tissues. Competition experiments using the adenosine receptor agonists R-N(6)-phenylisopropyladenosine, 5'-N-ethylcarboxamidoadenosine, and S-N(6)-phenylisopropyladenosine gave monophasic displacement curves with Ki values of 50 nM, 440 nM, and 4,300 nM, which agreed well with the GTP-inducible low affinity state in cardiac membranes. The low affinity for agonists was not due to agonist-induced desensitization, and correlated well with the corresponding IC50 values for the inhibition of cyclic AMP accumulation by isoprenaline. It is suggested that only a low affinity state of A1 receptors can be detected in intact rat myocytes due to the presence of high concentrations of guanine nucleotides in intact cells.