Skip to main content
SpringerLink
Log in

Decrease of serum adenine nucleotide hydrolysis in an irritant contact dermatitis mice model: potential P2X7R involvement

  • Published:
Molecular and Cellular Biochemistry Aims and scope Submit manuscript

Abstract

Extracellular adenosine 5′-triphosphate (ATP) has significant effects on a variety of pathological conditions and it is the main physiological agonist of P2X7 purinergic receptor (P2X7R). It is known that ATP acting via purinergic receptors plays a relevant role on skin inflammation, and P2X7R is required to neutrophil recruitment in a mice model of irritant contact dermatitis (ICD).The present study investigated the effects of chemical irritant croton oil (CrO) upon ATP, ADP, and AMP hydrolysis in mice blood serum, and the potential involvement of P2X7R. The topical application CrO induced a decrease on soluble ATP/ADPase activities (~50 %), and the treatment with the selective P2X7R antagonist, A438079, reversed these effects to control level. Furthermore, we showed that CrO decreased cellular viability (52.6 % ± 3.9) in relation to the control and caused necrosis in keratinocytes (PI positive cells). The necrosis induced by CrO was prevented by the pre-treatment with the selective P2X7R antagonist A438079. The results presented herein suggest that CrO exerts an inhibitory effect on the activity of ATPDase in mouse serum, reinforcing the idea that ICD has a pathogenic mechanism dependent of CD39. Furthermore, it is tempting to suggest that P2X7R may act as a controller of the extracellular levels of ATP.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3

References

  1. Eltzschig HK, Sitkovsky MV, Robson SC (2012) Purinergic signaling during inflammation. N Engl J Med 367:2322–2333

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  2. Burnstock G (2006) Historical review: ATP as a neurotransmitter. Trends Pharmacol Sci 27:166–176

    Article  CAS  PubMed  Google Scholar 

  3. Di Virgilio F, Boeynaems JM, Robson SC (2009) Extracellular nucleotides as negative modulators of immunity. Curr Opin Pharmacol 9:507–513

    Article  PubMed Central  PubMed  Google Scholar 

  4. Deaglio S, Robson SC (2011) Ectonucleotidases as regulators of purinergic signaling in thrombosis, inflammation, and immunity. Adv Pharmacol 61:301–332

    Article  CAS  PubMed  Google Scholar 

  5. Rittiner JE, Korboukh I, Hull-Ryde EA et al (2012) AMP is an adenosine A1 receptor agonist. J Biol Chem 287:5301–5309. doi:10.1074/jbc.M111.291666

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  6. Zimmermann H, Zebisch M, Sträter N (2012) Cellular function and molecular structure of ecto-nucleotidases. Purinergic Signal 8:437–502

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  7. Robson SC, Sévigny J, Zimmermann H (2006) The E-NTPDase family of ectonucleotidases: structure function relationships and pathophysiological significance. Purinergic Signal 2:409–430

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  8. Mizumoto N, Mummert ME, Shalhevet D, Takashima A (2003) Keratinocyte ATP release assay for testing skin-irritating potentials of structurally diverse chemicals. J Invest Dermatol 121:1066–1072

    Article  CAS  PubMed  Google Scholar 

  9. Pastore S, Mascia F, Gulinelli S et al (2007) Stimulation of purinergic receptors modulates chemokine expression in human keratinocytes. J Invest Dermatol 127:660–667. doi:10.1038/sj.jid.5700591

    Article  CAS  PubMed  Google Scholar 

  10. Ferrari D, Pizzirani C, Adinolfi E et al (2006) The P2X7 receptor: a key player in IL-1 processing and release. J Immunol 176:3877–3883

    Article  CAS  PubMed  Google Scholar 

  11. Da Silva GL, Sperotto NDM, Borges TJ et al (2013) P2X7 receptor is required for neutrophil accumulation in a mouse model of irritant contact dermatitis. Exp Dermatol 22:184–188

    Article  PubMed  Google Scholar 

  12. Han MH, Yoon WK, Lee H et al (2007) Topical application of silymarin reduces chemical-induced irritant contact dermatitis in BALB/c mice. Int Immunopharmacol 7:1651–1658

    Article  CAS  PubMed  Google Scholar 

  13. Nosbaum A, Vocanson M, Rozieres A et al (2009) Allergic and irritant contact dermatitis. Eur J Dermatol 19:325–332

    CAS  PubMed  Google Scholar 

  14. Mizumoto N, Kumamoto T, Robson SC et al (2002) CD39 is the dominant Langerhans cell-associated ecto-NTPDase: modulatory roles in inflammation and immune responsiveness. Nat Med 8:358–365

    Article  CAS  PubMed  Google Scholar 

  15. Heine P, Braun N, Heilbronn A, Zimmermann H (1999) Functional characterization of rat ecto-ATPase and ecto-ATP diphosphohydrolase after heterologous expression in CHO cells. Eur J Biochem 262:102–107

    Article  CAS  PubMed  Google Scholar 

  16. Zimmermann H (2001) Ectonucleotidases: Some recent developments and a note on nomenclature. Drug Dev Res 52:44–56

    Article  CAS  Google Scholar 

  17. Zimmermann M (1986) Ethical considerations in relation to pain in animal experimentation. Acta Physiol Scand Suppl 554:221–233

    CAS  PubMed  Google Scholar 

  18. Morrone FB, Horn AP, Stella J et al (2005) Increased resistance of glioma cell lines to extracellular ATP cytotoxicity. J Neurooncol 71:135–140

    Article  CAS  PubMed  Google Scholar 

  19. Bradford M (1976) Rapid and sensitive method for quantification of microgram quantities of protein utilizing principle of protein-dye-binding. Anal Biochem 72:248–254

    Article  CAS  PubMed  Google Scholar 

  20. Bavaresco L, Bernardi A, Braganhol E et al (2007) Dexamethasone inhibits proliferation and stimulates ecto-5′-nucleotidase/CD73 activity in C6 rat glioma cell line. J Neurooncol 84:1–8

    Article  CAS  PubMed  Google Scholar 

  21. Longhi MS, Robson SC, Bernstein SH et al (2013) Biological functions of ecto-enzymes in regulating extracellular adenosine levels in neoplastic and inflammatory disease states. J Mol Med 91:165–172

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  22. Bours MJL, Swennen ELR, Di Virgilio F et al (2006) Adenosine 5′-triphosphate and adenosine as endogenous signaling molecules in immunity and inflammation. Pharmacol Ther 112:358–404

    Article  CAS  PubMed  Google Scholar 

  23. Csóka BHG (2015) CD39 improves survival in microbial sepsis by attenuating systemic inflammation. FASEB J 1:25–36. doi:10.1096/fj.14-253567

    Article  Google Scholar 

  24. Morandini C-SR (2014) The role of P2X7 receptor in infectious inflammatory diseases and the influence of ectonucleotidases. Biomed J 2:169–177. doi:10.4103/2319-4170.127803

    Google Scholar 

  25. McDonald B, Pittman K, Menezes GB et al (2010) Intravascular danger signals guide neutrophils to sites of sterile inflammation. Science 330:362–366

    Article  CAS  PubMed  Google Scholar 

  26. Salmi M, Jalkanen S (2005) Cell-surface enzymes in control of leukocyte trafficking. Nat Rev Immunol 5:760–771

    Article  CAS  PubMed  Google Scholar 

  27. Carroll WA, Donnelly-Roberts D, Jarvis MF (2009) Selective P2X7 receptor antagonists for chronic inflammation and pain. Purinergic Signal 5:63–73

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  28. Boeynaems J-M, Communi D (2006) Modulation of inflammation by extracellular nucleotides. J Invest Dermatol 126:943–944

    Article  CAS  PubMed  Google Scholar 

  29. Ho CL, Yang CY, Lin WJ, Lin CH (2013) Ecto-nucleoside triphosphate diphosphohydrolase 2 modulates local atp-induced calcium signaling in human HaCaT keratinocytes. PLoS ONE 8:e57666

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  30. Burrell HE, Wlodarski B, Foster BJ et al (2005) Human keratinocytes release ATP and utilize three mechanisms for nucleotide interconversion at the cell surface. J Biol Chem 280:29667–29676

    Article  CAS  PubMed  Google Scholar 

  31. Welss T, Basketter D, Schröder K (2004) In vitro skin irritation: facts and future. State of the art review of mechanisms and models. Toxicol Vitro 18:231–243. doi:10.1016/j.tiv.2003.09.009

    Article  CAS  Google Scholar 

Download references

Acknowledgments

The authors thank Mr. Juliano Soares for the excellent technical assistance, Dr. Décio dos Santos Pinto Jr. for the donation of the cell line, Dr. Maria Martha Campos for her critical advice, and Me. Marina Petersen Gehring for helping with the experiments. Da Silva, G.L. thanks Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) for the fellowship.

Author information

Author notes

    Authors and Affiliations

    1. Faculdade de Farmácia, Pontificia Universidade Catolica do Rio Grande do Sul - PUCRS, Porto Alegre, RS, Brazil

      T. Erig, N. D. M. Sperotto & Fernanda Bueno Morrone

    2. Programa de Pós-Graduação em Biologia Celular e Molecular, Pontificia Universidade Catolica do Rio Grande do Sul - PUCRS, Porto Alegre, RS, Brazil

      R. F. Zanin, G. L. da Silva & Fernanda Bueno Morrone

    3. Instituto de Toxicologia e Farmacologia, Pontificia Universidade Catolica do Rio Grande do Sul - PUCRS, Porto Alegre, RS, Brazil

      R. F. Zanin, C. E. Leite & Fernanda Bueno Morrone

    4. Instituto de Biofisica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro - UFRJ, Rio de Janeiro, RJ, Brazil

      R. Coutinho-Silva

    5. Departamento de Bioquímica, Instituto de Ciências Básicas da Saúde, UFRGS, Porto Alegre, RS, Brazil

      A. M. O. Batastini

    6. Laboratório de Farmacologia Aplicada, Pontificia Universidade Catolica do Rio Grande do Sul, Avenida Ipiranga, 6681, Partenon, Porto Alegre, RS, 90619-900, Brazil

      Fernanda Bueno Morrone

    Authors
    1. R. F. Zanin
      View author publications

      You can also search for this author in PubMed Google Scholar

    2. G. L. da Silva
      View author publications

      You can also search for this author in PubMed Google Scholar

    3. T. Erig
      View author publications

      You can also search for this author in PubMed Google Scholar

    4. N. D. M. Sperotto
      View author publications

      You can also search for this author in PubMed Google Scholar

    5. C. E. Leite
      View author publications

      You can also search for this author in PubMed Google Scholar

    6. R. Coutinho-Silva
      View author publications

      You can also search for this author in PubMed Google Scholar

    7. A. M. O. Batastini
      View author publications

      You can also search for this author in PubMed Google Scholar

    8. Fernanda Bueno Morrone
      View author publications

      You can also search for this author in PubMed Google Scholar

    Corresponding author

    Correspondence to Fernanda Bueno Morrone.

    Additional information

    R. F. Zanin and G. L. da Silva have contributed equally.

    Rights and permissions

    Reprints and permissions

    About this article

    Check for updates. Verify currency and authenticity via CrossMark

    Cite this article

    Zanin, R.F., da Silva, G.L., Erig, T. et al. Decrease of serum adenine nucleotide hydrolysis in an irritant contact dermatitis mice model: potential P2X7R involvement. Mol Cell Biochem 404, 221–228 (2015). https://doi.org/10.1007/s11010-015-2381-7

    Download citation

    • Received:

    • Accepted:

    • Published:

    • Issue Date:

    • DOI: https://doi.org/10.1007/s11010-015-2381-7

    Keywords

    Search

    Navigation