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Cardiotonic steroid-resistant α1-Na+,K+-ATPase rescues renal epithelial cells from the cytotoxic action of ouabain: evidence for a Na +i ,K +i -independent mechanism

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Abstract

Mechanisms underlying the tissue-specific impact of cardiotonic steroids (CTS) on cell survival and death remain poorly understood. This study examines the role of Na+,K+-ATPase α subunits in death of Madin-Darby canine kidney (MDCK) cells evoked by 24-h exposure to ouabain. MDCK cells expressing a variant of the α1 isoform, CTS-sensitive α1S, were stably transfected with a cDNA encoding CTS-resistant α1R-Na+,K+-ATPase, whose expression was confirmed by RT–PCR. In mock-transfected and α1R-cells, maximal inhibition of 86Rb influx was observed at 10 and 1000 μM ouabain, respectively, thus confirming high abundance of α1R-Na+,K+-ATPase in these cells. Six-hour treatment of α1R-cells with 1000 μM ouabain led to the same elevation of the [Na+]i/[K+]i ratio that was detected in mock-transfected cells treated with 3 μM ouabain. However, in contrast to the massive death of mock-transfected cells exposed to 3 μM ouabain, α1R-cells survived after 24-h incubation with 1000 μM ouabain. Inversion of the [Na+]i/[K+]i ratio evoked by Na+,K+-ATPase inhibition in K+-free medium did not affect survival of α1R-cells but increased their sensitivity to ouabain. Our results show that the α1R subunit rescues MDCK cells from the cytotoxic action of CTS independently of inhibition of Na+,K+-ATPase-mediated Na+ and K+ fluxes and inversion of the [Na+]i/[K+]i ratio.

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Acknowledgments

This work was supported by grants from the Canadian Institutes of Health Research (MOP-81392 to SNO, JT and PH), Heart and Stroke Foundation of Quebec (to SNO, JT and PH) and Heart and Stroke Foundation of Ontario (NA-6324 to JWV) Canada. OA is a recipient of a fellowship from the Kidney Foundation of Canada.

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Authors and Affiliations

  1. Centre de recherche, Centre hospitalier de l’Université de Montréal (CHUM), Technopôle Angus, 2901 Rachel Est, Montreal, QC, H1W 4A4, Canada

    Olga A. Akimova, Johanne Tremblay, Pavel Hamet & Sergei N. Orlov

  2. Departments of Medicine and Biochemistry, University of Ottawa, Ottawa, ON, Canada

    James W. Van Huysse

  3. Faculty of Biology, M.V. Lomonosov Moscow State University, Moscow, Russia

    Olga A. Akimova & Sergei N. Orlov

  4. Institute of General Pathology and Pathophysiology, Russian Academy of Medical Sciences, Moscow, Russia

    Sergei N. Orlov

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  1. Olga A. Akimova
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  2. Johanne Tremblay
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  3. James W. Van Huysse
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  4. Pavel Hamet
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  5. Sergei N. Orlov
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Correspondence to Olga A. Akimova.

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Akimova, O.A., Tremblay, J., Van Huysse, J.W. et al. Cardiotonic steroid-resistant α1-Na+,K+-ATPase rescues renal epithelial cells from the cytotoxic action of ouabain: evidence for a Na +i ,K +i -independent mechanism. Apoptosis 15, 55–62 (2010). https://doi.org/10.1007/s10495-009-0429-4

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