Oxidant-induced cardiomyocyte injury: Identification of the cytoprotective effect of a dopamine 1 receptor agonist using a cell-based high-throughput assay

  • Authors:
    • Domokos Gerö
    • Katalin Módis
    • Nóra Nagy
    • Petra Szoleczky
    • Zoltán Dóri Tóth
    • György Dormán
    • Csaba Szabó
  • View Affiliations

  • Published online on: November 1, 2007     https://doi.org/10.3892/ijmm.20.5.749
  • Pages: 749-761
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Abstract

Myocyte injury due to myocardial reperfusion injury plays a crucial role in the pathogenesis of acute myocardial infarction even after successful coronary revascularization. Identification of compounds that reduce reperfusion-associated myocyte death is important. Therefore, we developed an in vitro model of myocardial reperfusion injury in H9c2 rat cardiomyocytes and applied a cell-based high-throughput approach to screen a standard library of pharmacologically active compounds (LOPAC) in order to identify drugs with cardioprotective effects. Oxidative stress was induced with hydrogen peroxide (H2O2) treatment, which resulted in ≈50% reduction in cell viability. Test compounds were added at a 3-µM final concentration as a pretreatment or in a delayed fashion (30 min after the peroxide challenge in order to imitate pharmacological treatment following angioplasty). Cells were cultured for 3 or 24 h. Viability was quantitated with the methylthiazolyldiphenyl-tetrazolium bromide method. Cytotoxicity and cytoprotection were also evaluated by measuring the lactate dehydrogenase activity in the cell culture supernatant. The screening identified a number of compounds with cytoprotective action, including molecules that are known to interfere with components of DNA repair and cell cycle progression, e.g. poly(ADP-ribose) polymerase (PARP) inhibitors, topoisomerase inhibitors, and cyclin dependent kinase inhibitors, or reduce energy consumption by interfering with cardiac myofilament function. A number of dopamine D1 receptor agonists also provided significant cytoprotection at 3 h, but only three of them showed a similar effect at 24 h: chloro- and bromo-APB and chloro-PB hydrobromide. Chloro-APB hydrobromide significantly reduced peroxide-induced PARP activation in the myocytes independently of its action on dopamine D1 receptors, but lacked PARP inhibitor capacity in a cell-free PARP assay system. In conclusion, the pattern of cytoprotective drugs identified in the current assay supports the overall validity of our model system. The findings demonstrate that cytoprotective agents, including novel indirect inhibitors of cellular PARP activation can be identified with the method, chloro-APB hydrobromide being one such compound. The current experimental setting can be employed for cell-based high-throughput screening of various compound libraries.

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november 2007
Volume 20 Issue 5

Print ISSN: 1107-3756
Online ISSN:1791-244X

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Spandidos Publications style
Gerö D, Módis K, Nagy N, Szoleczky P, Tóth ZD, Dormán G and Szabó C: Oxidant-induced cardiomyocyte injury: Identification of the cytoprotective effect of a dopamine 1 receptor agonist using a cell-based high-throughput assay. Int J Mol Med 20: 749-761, 2007.
APA
Gerö, D., Módis, K., Nagy, N., Szoleczky, P., Tóth, Z.D., Dormán, G., & Szabó, C. (2007). Oxidant-induced cardiomyocyte injury: Identification of the cytoprotective effect of a dopamine 1 receptor agonist using a cell-based high-throughput assay. International Journal of Molecular Medicine, 20, 749-761. https://doi.org/10.3892/ijmm.20.5.749
MLA
Gerö, D., Módis, K., Nagy, N., Szoleczky, P., Tóth, Z. D., Dormán, G., Szabó, C."Oxidant-induced cardiomyocyte injury: Identification of the cytoprotective effect of a dopamine 1 receptor agonist using a cell-based high-throughput assay". International Journal of Molecular Medicine 20.5 (2007): 749-761.
Chicago
Gerö, D., Módis, K., Nagy, N., Szoleczky, P., Tóth, Z. D., Dormán, G., Szabó, C."Oxidant-induced cardiomyocyte injury: Identification of the cytoprotective effect of a dopamine 1 receptor agonist using a cell-based high-throughput assay". International Journal of Molecular Medicine 20, no. 5 (2007): 749-761. https://doi.org/10.3892/ijmm.20.5.749