Abstract
P2 purinoceptor modulation of injury during ischemia-reperfusion was studied in murine hearts. Effects of P2 agonism or antagonism, and interstitial accumulation of P2 agonists (UTP, ATP, and ADP), were assessed in Langendorff perfused hearts during 20 min of ischemia and 45 min of reperfusion. In control hearts, ventricular pressure development recovered to 68 ± 4 mm Hg (63 ± 3% baseline), diastolic pressure remained elevated (23 ± 2 mm Hg), and 26 ± 4 U/g lactate dehydrogenase (LDH) was released during reperfusion, evidencing necrosis. Treatment with 250 nM UTP improved pressure development (85 ± 5 mm Hg, or 77 ± 2%) and reduced diastolic contracture (by ∼70%, to 7 ± 1 mm Hg) and LDH loss (by ∼60%, to 11 ± 2 U/g). In contrast, P2Y1 agonism with 50 nM 2-methyl-thio-ATP (2-MeSATP) was ineffective. In the presence of the P2Y antagonist suramin (10 or 200 μM), UTP no longer improved postischemic outcomes. Ischemia also substantially elevated interstitial [UTP], [ATP], and [ADP], potentially activating P2 receptors. This was supported in part by effects of antagonists: 200 μM suramin worsened LDH efflux (53 ± 9 IU/g) and contractile dysfunction (41 ± 2 mm Hg diastolic pressure; 28 ± 3 mm Hg developed pressure), as did P2Y antagonism with either 10 or 100 μM reactive blue 2. However, a 10 μM concentration of suramin failed to alter outcome. P2X antagonism with 10 μM pyridoxal phosphate-6-azo-(benzene-2,4-disulfonic acid and P2X1-selective pyridoxal-α5-phosphate-6-phenylazo-4′-carboxylic acid (MRS2159) (30 μM) was ineffective. Data collectively support cardioprotection with low concentrations of UTP, and they are consistent with P2Y2 involvement. Endogenous nucleotides may also play a protective role, as evidenced by effects of P2 antagonists, although this warrants further investigation.
Footnotes
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This work was supported by grants from the National Health and Medical Research Council of Australia and the National Heart Foundation of Australia. We gratefully acknowledge scholarship support for S.W. (Australian Postgraduate Award), and fellowship support for J.P.H. (National Heart Foundation of Australia) and J.N.P. (National Health and Medical Research Council of Australia).
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Article, publication date, and citation information can be found at http://jpet.aspetjournals.org.
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doi:10.1124/jpet.107.125815.
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ABBREVIATIONS: RB-2, reactive blue 2; PPADS, pyridoxal phosphate-6-azophenyl-2′, 4′-disulfonic acid; 2-MeSATP, 2-methylthio-ATP; LDH, lactate dehydrogenase; LV, left ventricular; MRS2159, pyridoxal-α5-phosphate-6-phenylazo-4′-carboxylic acid.
- Received May 15, 2007.
- Accepted September 7, 2007.
- The American Society for Pharmacology and Experimental Therapeutics
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