Extracellular nucleotide derivatives protect cardiomyocytes against hypoxic stress

O. Golan; Y. Issan; A. Isak; J. Leipziger; B. Robaye; A. Shainberg

doi:10.1016/j.bcp.2011.02.007

Extracellular nucleotide derivatives protect cardiomyocytes against hypoxic stress

O. Golan, Y. Issan, A. Isak, J. Leipziger, B. Robaye, A. Shainberg

The Mina and Everard Goodman Faculty of Life Sciences - at Bar-Ilan University

Research output: Contribution to journal › Article › peer-review

14 Scopus citations

Abstract

Rationale: Extracellular nucleotides have widespread effects and various cell responses. Whereas the effect of a purine nucleotide (ATP) and a pyrimidine nucleotide (UTP) on myocardial infarction has been examined, the role of different purine and pyrimidine nucleotides and nucleosides in cardioprotection against hypoxic stress has not been reported. Objective: To investigate the role of purine and pyrimidine nucleotides and nucleosides in protective effects in cardiomyocytes subjected to hypoxia. Methods and results: Rat cultured cardiomyocytes were treated with various extracellular nucleotides and nucleosides, before or during hypoxic stress. The results revealed that GTP or CTP exhibit cardioprotective ability, as revealed by lactate dehydrogenase (LDH) release, by propidium iodide (PI) staining, by cell morphology, and by preserved mitochondrial activity. Pretreatment with various P2 antagonists (suramin, RB-2, or PPADS) did not abolish the cardioprotective effect of the nucleotides. Moreover, P2Y₂^-/-_, P2Y ₄^-/-, and P2Y₂^-/-/P2Y ₄^-/- receptor knockouts mouse cardiomyocytes were significantly protected against hypoxic stress when treated with UTP. These results indicate that the protective effect is not mediated via those receptors. We found that a wide variety of triphosphate and diphosphate nucleotides (TTP, ITP, deoxyGTP, and GDP), provided significant cardioprotective effect. GMP, guanosine, and ribose phosphate provided no cardioprotective effect. Moreover, we observed that tri/di-phosphate alone assures cardioprotection. Treatment with extracellular nucleotides, or with tri/di-phosphate, administered under normoxic conditions or during hypoxic conditions, led to a decrease in reactive oxygen species production. Conclusions: Extracellular tri/di-phosphates are apparently the molecule responsible for cardioprotection against hypoxic damage, probably by preventing free radicals formation.

Original language	English
Pages (from-to)	1219-1227
Number of pages	9
Journal	Biochemical Pharmacology
Volume	81
Issue number	10
DOIs	https://doi.org/10.1016/j.bcp.2011.02.007
State	Published - 15 May 2011

Bibliographical note

Funding Information:
The authors thank Dr. Beverly Koller (University of North Carolina at Chapel Hill, Chapel Hill, NC) for supplying breeders of mice deficient in P2Y 2 -Receptor and the corresponding WT mice. The authors are indebted to Avrille Goldreich for helping to prepare and submitting this manuscript for publication. This work was in part supported by The Adar Program for the Advancement of Research in Heart Function and the Horowitz Foundation at Bar-Ilan University .

Funding

The authors thank Dr. Beverly Koller (University of North Carolina at Chapel Hill, Chapel Hill, NC) for supplying breeders of mice deficient in P2Y 2 -Receptor and the corresponding WT mice. The authors are indebted to Avrille Goldreich for helping to prepare and submitting this manuscript for publication. This work was in part supported by The Adar Program for the Advancement of Research in Heart Function and the Horowitz Foundation at Bar-Ilan University .

Funders	Funder number
Horowitz Foundation at Bar-Ilan University

Keywords

Cardioprotection
Purines
Pyrimidines
Pyrophosphate
ROS

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10.1016/j.bcp.2011.02.007

Cite this

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title = "Extracellular nucleotide derivatives protect cardiomyocytes against hypoxic stress",

abstract = "Rationale: Extracellular nucleotides have widespread effects and various cell responses. Whereas the effect of a purine nucleotide (ATP) and a pyrimidine nucleotide (UTP) on myocardial infarction has been examined, the role of different purine and pyrimidine nucleotides and nucleosides in cardioprotection against hypoxic stress has not been reported. Objective: To investigate the role of purine and pyrimidine nucleotides and nucleosides in protective effects in cardiomyocytes subjected to hypoxia. Methods and results: Rat cultured cardiomyocytes were treated with various extracellular nucleotides and nucleosides, before or during hypoxic stress. The results revealed that GTP or CTP exhibit cardioprotective ability, as revealed by lactate dehydrogenase (LDH) release, by propidium iodide (PI) staining, by cell morphology, and by preserved mitochondrial activity. Pretreatment with various P2 antagonists (suramin, RB-2, or PPADS) did not abolish the cardioprotective effect of the nucleotides. Moreover, P2Y2-/-, P2Y 4-/-, and P2Y2-/-/P2Y 4-/- receptor knockouts mouse cardiomyocytes were significantly protected against hypoxic stress when treated with UTP. These results indicate that the protective effect is not mediated via those receptors. We found that a wide variety of triphosphate and diphosphate nucleotides (TTP, ITP, deoxyGTP, and GDP), provided significant cardioprotective effect. GMP, guanosine, and ribose phosphate provided no cardioprotective effect. Moreover, we observed that tri/di-phosphate alone assures cardioprotection. Treatment with extracellular nucleotides, or with tri/di-phosphate, administered under normoxic conditions or during hypoxic conditions, led to a decrease in reactive oxygen species production. Conclusions: Extracellular tri/di-phosphates are apparently the molecule responsible for cardioprotection against hypoxic damage, probably by preventing free radicals formation.",

keywords = "Cardioprotection, Purines, Pyrimidines, Pyrophosphate, ROS",

author = "O. Golan and Y. Issan and A. Isak and J. Leipziger and B. Robaye and A. Shainberg",

note = "Funding Information: The authors thank Dr. Beverly Koller (University of North Carolina at Chapel Hill, Chapel Hill, NC) for supplying breeders of mice deficient in P2Y 2 -Receptor and the corresponding WT mice. The authors are indebted to Avrille Goldreich for helping to prepare and submitting this manuscript for publication. This work was in part supported by The Adar Program for the Advancement of Research in Heart Function and the Horowitz Foundation at Bar-Ilan University .",

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AU - Golan, O.

AU - Issan, Y.

AU - Isak, A.

AU - Leipziger, J.

AU - Robaye, B.

AU - Shainberg, A.

N1 - Funding Information: The authors thank Dr. Beverly Koller (University of North Carolina at Chapel Hill, Chapel Hill, NC) for supplying breeders of mice deficient in P2Y 2 -Receptor and the corresponding WT mice. The authors are indebted to Avrille Goldreich for helping to prepare and submitting this manuscript for publication. This work was in part supported by The Adar Program for the Advancement of Research in Heart Function and the Horowitz Foundation at Bar-Ilan University .

PY - 2011/5/15

Y1 - 2011/5/15

N2 - Rationale: Extracellular nucleotides have widespread effects and various cell responses. Whereas the effect of a purine nucleotide (ATP) and a pyrimidine nucleotide (UTP) on myocardial infarction has been examined, the role of different purine and pyrimidine nucleotides and nucleosides in cardioprotection against hypoxic stress has not been reported. Objective: To investigate the role of purine and pyrimidine nucleotides and nucleosides in protective effects in cardiomyocytes subjected to hypoxia. Methods and results: Rat cultured cardiomyocytes were treated with various extracellular nucleotides and nucleosides, before or during hypoxic stress. The results revealed that GTP or CTP exhibit cardioprotective ability, as revealed by lactate dehydrogenase (LDH) release, by propidium iodide (PI) staining, by cell morphology, and by preserved mitochondrial activity. Pretreatment with various P2 antagonists (suramin, RB-2, or PPADS) did not abolish the cardioprotective effect of the nucleotides. Moreover, P2Y2-/-, P2Y 4-/-, and P2Y2-/-/P2Y 4-/- receptor knockouts mouse cardiomyocytes were significantly protected against hypoxic stress when treated with UTP. These results indicate that the protective effect is not mediated via those receptors. We found that a wide variety of triphosphate and diphosphate nucleotides (TTP, ITP, deoxyGTP, and GDP), provided significant cardioprotective effect. GMP, guanosine, and ribose phosphate provided no cardioprotective effect. Moreover, we observed that tri/di-phosphate alone assures cardioprotection. Treatment with extracellular nucleotides, or with tri/di-phosphate, administered under normoxic conditions or during hypoxic conditions, led to a decrease in reactive oxygen species production. Conclusions: Extracellular tri/di-phosphates are apparently the molecule responsible for cardioprotection against hypoxic damage, probably by preventing free radicals formation.

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Extracellular nucleotide derivatives protect cardiomyocytes against hypoxic stress

Abstract

Bibliographical note

Funding

Keywords

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