Preconditioning of rat hearts by adenosine A1 or A3 receptor activation

doi:10.1016/S0014-2999(01)01611-9

European Journal of Pharmacology

Volume 441, Issue 3, 26 April 2002, Pages 165-172

https://doi.org/10.1016/S0014-2999(01)01611-9 Get rights and content

Abstract

Our study in rat hearts examined whether activation of adenosine A₁ or A₃ receptors improved functional recovery and reduced apoptosis resulting from low-flow ischemia. Prior to 30 min low-flow ischemia (0.6 ml/min; 6% of baseline flow), Langendorff rat hearts were preconditioned with two 5-min cycles of (a) ischemia (PC; n=7), (b) infusion of 250 nM adenosine A₁ receptor agonist 2-chloro-N⁶-cyclopentyladenosine (CCPA; n=6), or (c) infusion of 50 nM adenosine A₃ receptor agonist N⁶-(3-iodobenzyl)-adenosine-5′-N-methyl-uronamide (IB-MECA; n=8). Recovery of function was improved in PC (71±3%), CCPA (68±6%) and IB-MECA (68±4%) groups compared to control hearts (46±5%; P<0.05). Cumulative release of total purines during ischemia–reperfusion was approx. 50% lower in PC, CCPA and IB-MECA groups compared to controls (P<0.05) and was significantly correlated to the percentage functional recovery (R²=0.55; P<0.05). The number of cytosolic histone-associated-DNA fragments, a hallmark of apoptosis and measured by Enzyme Linked ImmunoSorbent Assay (ELISA), was small and not different between groups after 30 min reperfusion. However, CCPA (0.6±0.1 absorbance units) and MECA (0.7±0.1 units; P<0.05 vs. PC) decreased apoptosis after 150 min reperfusion compared to PC (1.4±0.3 units) and control (1.2±0.1 units) hearts. This study shows that adenosine triggers protection of function in preconditioned rat hearts via both the adenosine A₁ and A₃ receptor. In clinical practice, pharmacological stimulation of adenosine A₃ receptors may be advantageous over adenosine A₁ receptor activation due to a lack of contractile side-effects. In contrast to ischemic preconditioning, pharmacological stimulation of adenosine A₁ or A₃ receptors reduced apoptosis. Furthermore, total purine release may serve as a marker of the degree of functional protection.

Introduction

Ischemic preconditioning refers to the paradoxical mechanism that short, pre-emptive periods of ischemia protect the heart from a subsequent period of prolonged ischemia (Murry et al., 1986). This phenomenon seems to occur in all vertebrates including humans Cleveland et al., 1997, Yellon et al., 1993. Of the humoral factors released during its induction that may trigger the event, adenosine is one of the most important. In most animal species and humans Carr et al., 1997, Lee et al., 1995, Leesar et al., 1997, adenosine's cardioprotective effects are mediated via adenosine A₁ receptors located on the myocardial membrane. The recently discovered adenosine A₃ receptor has also been implicated in preconditioning of chick Liang and Jacobson, 1998, Stambaugh et al., 1997, Strickler et al., 1996, rabbit Auchampach et al., 1997b, Hill et al., 1998, Rice et al., 1996, Tracey et al., 1998, Wang et al., 1997, and human (Carr et al., 1997) hearts. However, adenosine-mediated cardioprotection is believed to play no role in rat hearts (Ganote and Armstrong, 2000). In contrast, we De Jonge et al., 1998, De Jonge et al., 2001 and others Ford et al., 1998, Headrick, 1996 observed beneficial effects of adenosine A₁ receptor stimulation in this species. To the best of our knowledge, no study has examined before the role of the adenosine A₃ receptor in preconditioning of rat hearts. However, a one study (Thourani et al., 1999) observed improved functional recovery and reduced creatine kinase release after 30 min no-flow ischemia in isolated rat hearts pretreated with a selective adenosine A₃ receptor agonist for 12 min without a washout period. Apart from reduced ventricular arrhythmias, infarct size, and contractile dysfunctioning, ischemic preconditioning has also been shown to reduce apoptosis in vitro (Gottlieb et al., 1996) and in vivo (Piot et al., 1997). No data exist as to whether ischemic preconditioning effects on apoptosis are also mimicked by stimulation of adenosine A₁/A₃ receptors prior to ischemia. Thus, this study examined the effectiveness of both adenosine A₁ and A₃ receptor stimulation in reducing contractile dysfunction and apoptosis in rat hearts after low-flow ischemia and reperfusion. This study shows that the adenosine A₃ receptor, in addition to the adenosine A₁ receptor, is involved in preconditioning of rat hearts. Although the degree of apoptosis was low, pharmacological preconditioning reduced its occurrence.

Section snippets

Methods

All animals were treated in conformation with the guiding principles in the care and use of animals as approved by the American Physiological Society. The Animal Welfare Committee, Erasmus University Rotterdam, approved the protocol.

Contractile function and coronary flow

Since preconditioning may affect recovery of developed pressure and heart rate to a different degree (Bradamante et al., 1993), the rate-force product will be presented in this study as a reliable index of contractile functioning. After 20 min stabilization, there were no differences between groups in rate-force product (mean and S.E.M. of all groups: 4510±224 g/min) or coronary flow rate (10±1 ml/min). Infusion of IB-MECA did not significantly affect preischemic rate-force product (Fig. 2) but

Role of the adenosine receptor in preconditioning of rat hearts

This is the first study in isolated rat hearts indicating that adenosine A₃ receptors are also involved in protection by ischemic preconditioning in rat hearts. Furthermore, in this study we confirm earlier observations of our group De Jonge and De Jong, 1999, De Jonge et al., 2001 that CCPA, a selective adenosine A₁ receptor agonist, reduces ischemic injury in rat hearts. Infusion of the selective adenosine A₃ receptor agonist IB-MECA prior to ischemia, resulted in a similar degree of

Conclusion

The present study demonstrates that adenosine A₁ and A₃ receptors are implicated in preconditioning of rat hearts. The degree of apoptosis was low in hearts subjected to 30 min underperfusion and reperfusion and was reduced by pharmacological preconditioning. Adenosine A₃ receptor agonists may represent a new, potentially useful therapeutic class of agents for providing cardioprotection as they lack cardiovascular side effects associated with adenosine A₁ receptor activation.

Acknowledgements

We gratefully acknowledge the support of the Netherlands Heart Foundation (NHS 94.043) and would like to thank Ms. E. Keijzer for analytical support. We also appreciate the stimulating discussions in the Biomed Concerted action ‘The New Ischemic Syndromes’ (Biomed II, BMH4-CT950838).

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Preconditioning of rat hearts by adenosine A1 or A3 receptor activation

Abstract

Introduction

Section snippets

Methods

Contractile function and coronary flow

Role of the adenosine receptor in preconditioning of rat hearts

Conclusion

Acknowledgements

Canine mast cell adenosine receptors: cloning and expression of the A3 receptor and evidence that degranulation is mediated by the A2B receptor

Mol. Pharmacol.

Selective activation of A3 adenosine receptors with N6-(3-iodobenzyl)adenosine-5′-N-methyluronamide protects against myocardial stunning and infarction without hemodynamic changes in conscious rabbits

Circ. Res.

Adenosine inhibits neutrophil degranulation in activated human whole blood: involvement of adenosine A2 and A3 receptors

J. Immunol.

Intracellular magnesium homeostasis is involved in the functional recovery of preconditioned rat heart

Biochim. Biophys. Res. Commun.

Evidence for a role for both the adenosine A1 and A3 receptors in protection of isolated human atrial muscle against simulated ischaemia

Cardiovasc. Res.

Improved functional recovery by ischaemic preconditioning is not mediated by adenosine in the globally ischaemic isolated rat heart

Cardiovasc. Res.

Ischemic preconditioning of human myocardium: protein kinase C mediates a permissive role for alpha1-adrenoceptors

Am. J. Physiol.

Ischemic preconditioning and glucose metabolism during low-flow ischemia: role of the adenosine A1 receptor

Cardiovasc. Res.

Carbohydrates and purines in underperfused hearts, protected by ischemic preconditioning

J. Mol. Cell. Cardiol.

Role of adenosine and glycogen in ischemic preconditioning of rat hearts

Eur. J. Pharmacol.

Adenosine alters glucose use during ischemia and reperfusion in isolated rat hearts

Circulation

Apoptosis in ischemic and reperfused rat myocardium

Circ. Res.

Intrinsic ANG II type 1 receptor stimulation contributes to recovery of postischemic mechanical function

Am. J. Physiol.

Mast cell degranulation following adenosine A3 receptor activation in rats

Eur. J. Pharmacol.

Adenosine and preconditioning in the rat heart

Cardiovasc. Res.

Attenuated purine production during subsequent ischemia in preconditioned rabbit myocardium is unrelated to the mechanism of protection

J. Mol. Cell. Cardiol.

Preconditioning rabbit cardiomyocytes: role of pH, vacuolar proton ATPase, and apoptosis

J. Clin. Invest.

Ischemic preconditioning: bioenergetic and metabolic changes and the role of endogenous adenosine

J. Mol. Cell. Cardiol.

Relative importance of adenosine A1 and A3 receptors in mediating physiological or pharmacological protection from ischemic myocardial injury in the rabbit heart

J. Mol. Cell. Cardiol.

Preconditioning cultured human pediatric myocytes requires adenosine and protein kinase C

Am. J. Physiol.

Apoptotic and necrotic myocyte cell deaths are independent contributing variables of infarct size in rats

Lab. Invest.

Adenosine A3 receptor stimulation inhibits migration of human eosinophils

J. Leukocyte Biol.

Species-dependent hemodynamic effects of adenosine A3-receptor agonists IB-MECA and Cl-IB-MECA

Am. J. Physiol.

Pretreatment of human myocardium with adenosine during open heart surgery

J. Cardiovasc. Surg.

Preconditioning of rat hearts by adenosine A₁ or A₃ receptor activation

Canine mast cell adenosine receptors: cloning and expression of the A₃ receptor and evidence that degranulation is mediated by the A_2B receptor

Selective activation of A₃ adenosine receptors with N⁶-(3-iodobenzyl)adenosine-5′-N-methyluronamide protects against myocardial stunning and infarction without hemodynamic changes in conscious rabbits

Adenosine inhibits neutrophil degranulation in activated human whole blood: involvement of adenosine A₂ and A₃ receptors

Evidence for a role for both the adenosine A₁ and A₃ receptors in protection of isolated human atrial muscle against simulated ischaemia

Ischemic preconditioning of human myocardium: protein kinase C mediates a permissive role for alpha₁-adrenoceptors

Ischemic preconditioning and glucose metabolism during low-flow ischemia: role of the adenosine A₁ receptor

Mast cell degranulation following adenosine A₃ receptor activation in rats

Relative importance of adenosine A₁ and A₃ receptors in mediating physiological or pharmacological protection from ischemic myocardial injury in the rabbit heart

Adenosine A₃ receptor stimulation inhibits migration of human eosinophils

Species-dependent hemodynamic effects of adenosine A₃-receptor agonists IB-MECA and Cl-IB-MECA