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Cell Biology and Toxicology

, Volume 32, Issue 3, pp 229–247 | Cite as

Extracellular ATP protects pancreatic duct epithelial cells from alcohol-induced damage through P2Y1 receptor-cAMP signal pathway

  • Jong Bae Seo
  • Seung-Ryoung Jung
  • Bertil Hille
  • Duk-Su Koh
Original Article

Abstract

Extracellular adenosine-5′-triphosphate (ATP) regulates cell death and survival of neighboring cells. The detailed effects are diverse depending on cell types and extracellular ATP concentration. We addressed the effect of ATP on ethanol-induced cytotoxicity in epithelial cells, the cell type that experiences the highest concentrations of alcohol. Using pancreatic duct epithelial cells (PDEC), we found that a micromolar range of ATP reverses all intracellular toxicity mechanisms triggered by exceptionally high doses of ethanol and, thus, improves cell viability dramatically. Out of the many purinergic receptors expressed in PDEC, the P2Y1 receptor was identified to mediate the protective effect, based on pharmacological and siRNA assays. Activation of P2Y1 receptors increased intracellular cyclic adenosine monophosphate (cAMP). The protective effect of ATP was mimicked by forskolin and 8-Br-cAMP but inhibited by a protein kinase A (PKA) inhibitor, H-89. Finally, ATP reverted leakiness of PDEC monolayers induced by ethanol and helped to maintain epithelial integrity. We suggest that purinergic receptors reduce extreme alcohol-induced cell damage via the cAMP signal pathway in PDEC and some other types of cells.

Keywords

Apoptosis Alcohol ATP cAMP Pancreatic duct epithelial cells Purinergic receptor 

Abbreviations

ADH

Alcohol dehydrogenase

ALDH

Acetaldehyde dehydrogenase

FAEE

Fatty acid ethyl ester

FRET

Fluorescence resonance energy transfer

GAPDH

Glyceraldehyde-3-phosphate dehydrogenase

MMP

Mitochondrial membrane potential

PDEC

Pancreatic duct epithelial cells

PKA

Protein kinase A

PKC

Protein kinase C

P2 receptor

Purinergic type 2 receptor

PMA

Phorbol 12-myristate 13-acetate

ROS

Reactive oxygen species

Notes

Acknowledgments

We thank Mark Moody and Lea M. Miller for technical assistance; Drs. Eamonn J. Dickson, and Toan N. Nguyen for helpful discussion and reading of manuscript: and Drs. Martin Kruse, Oscar Vivas, Haijie Yu, Po-Ni Lai, and Sei-Hum Jang for comments and helpful discussion. This work was supported by National Institutes of Health grant (R01-DK080840 to D.S.K.).

Author contributions

J.B.S., S.R.J., B.H., and D.S.K. developed the concept and designed research; J.B.S. and S.R.J. performed the experiments; J.B.S., S.R.J., and D.S.K. analyzed data; J.B.S., S.R.J., B.H., and D.S.K. interpreted results of experiments; J.B.S. and S.R.J. prepared figures; J.B.S. drafted manuscript; and J.B.S., S.R.J., B.H., and D.S.K. edited manuscript.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no competing interests

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Copyright information

© Springer Science+Business Media Dordrecht 2016

Authors and Affiliations

  • Jong Bae Seo
    • 1
    • 2
  • Seung-Ryoung Jung
    • 1
  • Bertil Hille
    • 1
  • Duk-Su Koh
    • 1
  1. 1.Department of Physiology and BiophysicsUniversity of WashingtonSeattleUSA
  2. 2.Department of Medicine, Division of Endocrinology and MetabolismUniversity of California San DiegoLa JollaUSA

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