Adenosine and inosine exert cytoprotective effects in an in vitro model of liver ischemia-reperfusion injury

Katalin Modis, Domokos Gero, Rita Stangl, Olivér Rosero, Attila Szijártó, Gábor Lotz, Petra Mohácsik, Petra Szoleczky, Ciro Coletta, Csaba Szabo

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

Liver ischemia represents a common clinical problem. In the present study, using an in vitro model of hepatic ischemia-reperfusion injury, we evaluated the potential cyto-protective effect of the purine metabolites, such as adenosine and inosine, and studied the mode of their pharmacological actions. The human hepatocellular carcinoma-derived cell line HepG2 was subjected to combined oxygen-glucose deprivation (COGD; 0-14-24 h), followed by re-oxygenation (0-4-24 h). Adenosine or inosine (300-1,000 μM) were applied in pretreatment. Cell viability and cytotoxicity were measured by the 3-(4,5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2H-tetrazolium bromide and lactate dehydrogenase methods, respectively. The results showed that both adenosine and inosine exerted cytoprotective effects, and these effects were not related to receptor-mediated actions, since they were not prevented by selective adenosine receptor antagonists. On the other hand, the adenosine deaminase inhibitor erythro-9-(2-hydroxy-3-nonyl) adenine hydrochloride (EHNA, 10 μM) markedly and almost fully reversed the protective effect of adenosine during COGD, while it did not influence the cyto-protective effect of inosine in the same assay conditions. These results suggest that the cytoprotective effects are related to intracellular actions, and, in the case of adenosine also involve intracellular conversion to inosine. The likely interpretation of these findings is that inosine serves as an alternative source of energy to produce ATP during hypoxic conditions. The protective effects are also partially dependent on adenosine kinase, as the inhibitor 4-amino-5-(3-bromophenyl)-7-(6-morpholino-pyridin-3-yl) pyrido[2,3-d]pyrimidine, 2HCl (ABT 702, 30 μM) significantly reversed the protective effect of both adenosine and inosine during hypoxia and re-oxygen-ation. Collectively, the current results support the view that during hypoxia, adenosine and inosine exert cytoprotective effects via receptor-independent, intracellular modes of action, which, in part, depend on the restoration of cellular bioenergetics. The present study supports the view that testing of inosine for protection against various forms of warm and cold liver ischemia is relevant.

Original languageEnglish (US)
Pages (from-to)437-446
Number of pages10
JournalInternational Journal of Molecular Medicine
Volume31
Issue number2
DOIs
StatePublished - Feb 2013

Fingerprint

Inosine
Reperfusion Injury
Adenosine
Liver
Adenosine Deaminase Inhibitors
Adenosine Kinase
Oxygen
Purinergic P1 Receptor Antagonists
Cold Ischemia
In Vitro Techniques
Morpholinos
L-Lactate Dehydrogenase
Energy Metabolism
Hepatocellular Carcinoma
Cell Survival
Ischemia
Adenosine Triphosphate
Pharmacology
Glucose
Cell Line

Keywords

  • Adenosine
  • Cytoprotection
  • Hepatocytes
  • Inosine
  • Ischemia-reperfusion
  • Liver

ASJC Scopus subject areas

  • Genetics

Cite this

Adenosine and inosine exert cytoprotective effects in an in vitro model of liver ischemia-reperfusion injury. / Modis, Katalin; Gero, Domokos; Stangl, Rita; Rosero, Olivér; Szijártó, Attila; Lotz, Gábor; Mohácsik, Petra; Szoleczky, Petra; Coletta, Ciro; Szabo, Csaba.

In: International Journal of Molecular Medicine, Vol. 31, No. 2, 02.2013, p. 437-446.

Research output: Contribution to journalArticle

Modis, Katalin ; Gero, Domokos ; Stangl, Rita ; Rosero, Olivér ; Szijártó, Attila ; Lotz, Gábor ; Mohácsik, Petra ; Szoleczky, Petra ; Coletta, Ciro ; Szabo, Csaba. / Adenosine and inosine exert cytoprotective effects in an in vitro model of liver ischemia-reperfusion injury. In: International Journal of Molecular Medicine. 2013 ; Vol. 31, No. 2. pp. 437-446.
@article{0da2d0e425cf4a678419c33e47001cfb,
title = "Adenosine and inosine exert cytoprotective effects in an in vitro model of liver ischemia-reperfusion injury",
abstract = "Liver ischemia represents a common clinical problem. In the present study, using an in vitro model of hepatic ischemia-reperfusion injury, we evaluated the potential cyto-protective effect of the purine metabolites, such as adenosine and inosine, and studied the mode of their pharmacological actions. The human hepatocellular carcinoma-derived cell line HepG2 was subjected to combined oxygen-glucose deprivation (COGD; 0-14-24 h), followed by re-oxygenation (0-4-24 h). Adenosine or inosine (300-1,000 μM) were applied in pretreatment. Cell viability and cytotoxicity were measured by the 3-(4,5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2H-tetrazolium bromide and lactate dehydrogenase methods, respectively. The results showed that both adenosine and inosine exerted cytoprotective effects, and these effects were not related to receptor-mediated actions, since they were not prevented by selective adenosine receptor antagonists. On the other hand, the adenosine deaminase inhibitor erythro-9-(2-hydroxy-3-nonyl) adenine hydrochloride (EHNA, 10 μM) markedly and almost fully reversed the protective effect of adenosine during COGD, while it did not influence the cyto-protective effect of inosine in the same assay conditions. These results suggest that the cytoprotective effects are related to intracellular actions, and, in the case of adenosine also involve intracellular conversion to inosine. The likely interpretation of these findings is that inosine serves as an alternative source of energy to produce ATP during hypoxic conditions. The protective effects are also partially dependent on adenosine kinase, as the inhibitor 4-amino-5-(3-bromophenyl)-7-(6-morpholino-pyridin-3-yl) pyrido[2,3-d]pyrimidine, 2HCl (ABT 702, 30 μM) significantly reversed the protective effect of both adenosine and inosine during hypoxia and re-oxygen-ation. Collectively, the current results support the view that during hypoxia, adenosine and inosine exert cytoprotective effects via receptor-independent, intracellular modes of action, which, in part, depend on the restoration of cellular bioenergetics. The present study supports the view that testing of inosine for protection against various forms of warm and cold liver ischemia is relevant.",
keywords = "Adenosine, Cytoprotection, Hepatocytes, Inosine, Ischemia-reperfusion, Liver",
author = "Katalin Modis and Domokos Gero and Rita Stangl and Oliv{\'e}r Rosero and Attila Szij{\'a}rt{\'o} and G{\'a}bor Lotz and Petra Moh{\'a}csik and Petra Szoleczky and Ciro Coletta and Csaba Szabo",
year = "2013",
month = "2",
doi = "10.3892/ijmm.2012.1203",
language = "English (US)",
volume = "31",
pages = "437--446",
journal = "International Journal of Molecular Medicine",
issn = "1107-3756",
publisher = "Spandidos Publications",
number = "2",

}

TY - JOUR

T1 - Adenosine and inosine exert cytoprotective effects in an in vitro model of liver ischemia-reperfusion injury

AU - Modis, Katalin

AU - Gero, Domokos

AU - Stangl, Rita

AU - Rosero, Olivér

AU - Szijártó, Attila

AU - Lotz, Gábor

AU - Mohácsik, Petra

AU - Szoleczky, Petra

AU - Coletta, Ciro

AU - Szabo, Csaba

PY - 2013/2

Y1 - 2013/2

N2 - Liver ischemia represents a common clinical problem. In the present study, using an in vitro model of hepatic ischemia-reperfusion injury, we evaluated the potential cyto-protective effect of the purine metabolites, such as adenosine and inosine, and studied the mode of their pharmacological actions. The human hepatocellular carcinoma-derived cell line HepG2 was subjected to combined oxygen-glucose deprivation (COGD; 0-14-24 h), followed by re-oxygenation (0-4-24 h). Adenosine or inosine (300-1,000 μM) were applied in pretreatment. Cell viability and cytotoxicity were measured by the 3-(4,5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2H-tetrazolium bromide and lactate dehydrogenase methods, respectively. The results showed that both adenosine and inosine exerted cytoprotective effects, and these effects were not related to receptor-mediated actions, since they were not prevented by selective adenosine receptor antagonists. On the other hand, the adenosine deaminase inhibitor erythro-9-(2-hydroxy-3-nonyl) adenine hydrochloride (EHNA, 10 μM) markedly and almost fully reversed the protective effect of adenosine during COGD, while it did not influence the cyto-protective effect of inosine in the same assay conditions. These results suggest that the cytoprotective effects are related to intracellular actions, and, in the case of adenosine also involve intracellular conversion to inosine. The likely interpretation of these findings is that inosine serves as an alternative source of energy to produce ATP during hypoxic conditions. The protective effects are also partially dependent on adenosine kinase, as the inhibitor 4-amino-5-(3-bromophenyl)-7-(6-morpholino-pyridin-3-yl) pyrido[2,3-d]pyrimidine, 2HCl (ABT 702, 30 μM) significantly reversed the protective effect of both adenosine and inosine during hypoxia and re-oxygen-ation. Collectively, the current results support the view that during hypoxia, adenosine and inosine exert cytoprotective effects via receptor-independent, intracellular modes of action, which, in part, depend on the restoration of cellular bioenergetics. The present study supports the view that testing of inosine for protection against various forms of warm and cold liver ischemia is relevant.

AB - Liver ischemia represents a common clinical problem. In the present study, using an in vitro model of hepatic ischemia-reperfusion injury, we evaluated the potential cyto-protective effect of the purine metabolites, such as adenosine and inosine, and studied the mode of their pharmacological actions. The human hepatocellular carcinoma-derived cell line HepG2 was subjected to combined oxygen-glucose deprivation (COGD; 0-14-24 h), followed by re-oxygenation (0-4-24 h). Adenosine or inosine (300-1,000 μM) were applied in pretreatment. Cell viability and cytotoxicity were measured by the 3-(4,5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2H-tetrazolium bromide and lactate dehydrogenase methods, respectively. The results showed that both adenosine and inosine exerted cytoprotective effects, and these effects were not related to receptor-mediated actions, since they were not prevented by selective adenosine receptor antagonists. On the other hand, the adenosine deaminase inhibitor erythro-9-(2-hydroxy-3-nonyl) adenine hydrochloride (EHNA, 10 μM) markedly and almost fully reversed the protective effect of adenosine during COGD, while it did not influence the cyto-protective effect of inosine in the same assay conditions. These results suggest that the cytoprotective effects are related to intracellular actions, and, in the case of adenosine also involve intracellular conversion to inosine. The likely interpretation of these findings is that inosine serves as an alternative source of energy to produce ATP during hypoxic conditions. The protective effects are also partially dependent on adenosine kinase, as the inhibitor 4-amino-5-(3-bromophenyl)-7-(6-morpholino-pyridin-3-yl) pyrido[2,3-d]pyrimidine, 2HCl (ABT 702, 30 μM) significantly reversed the protective effect of both adenosine and inosine during hypoxia and re-oxygen-ation. Collectively, the current results support the view that during hypoxia, adenosine and inosine exert cytoprotective effects via receptor-independent, intracellular modes of action, which, in part, depend on the restoration of cellular bioenergetics. The present study supports the view that testing of inosine for protection against various forms of warm and cold liver ischemia is relevant.

KW - Adenosine

KW - Cytoprotection

KW - Hepatocytes

KW - Inosine

KW - Ischemia-reperfusion

KW - Liver

UR - http://www.scopus.com/inward/record.url?scp=84873106319&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84873106319&partnerID=8YFLogxK

U2 - 10.3892/ijmm.2012.1203

DO - 10.3892/ijmm.2012.1203

M3 - Article

VL - 31

SP - 437

EP - 446

JO - International Journal of Molecular Medicine

JF - International Journal of Molecular Medicine

SN - 1107-3756

IS - 2

ER -