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Impaired mitochondrial functions contribute to 3-bromopyruvate toxicity in primary rat and mouse hepatocytes

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Abstract

A compound with promising anticancer properties, 3-bromopyruvate (3-BP) is a synthetic derivative of a pyruvate molecule; however, its toxicity in non-malignant cells has not yet been fully elucidated. Therefore, we elected to study the effects of 3-BP on primary hepatocytes in monolayer cultures, permeabilized hepatocytes and isolated mitochondria. After a 1-h treatment with 100 μM 3-BP cell viability of rat hepatocytes was decreased by 30 % as measured by the WST-1 test (p < 0.001); after 3-h exposure to ≥200 μM 3-BP lactate dehydrogenase leakage was increased (p < 0.001). Reactive oxygen species production was increased in the cell cultures after a 1-h treatment at concentrations ≥100 μmol/l (p < 0.01), and caspase 3 activity was increased after a 20-h incubation with 150 μM and 200 μM 3-BP (p < 0.001). This toxic effect of 3-BP was also proved using primary mouse hepatocytes. In isolated mitochondria, 3-BP induced a dose- and time-dependent decrease of mitochondrial membrane potential during a 10-min incubation both with Complex I substrates glutamate + malate or Complex II substrate succinate, although this decrease was more pronounced with the latter. We also measured the effect of 3-BP on respiration of isolated mitochondria. ADP-activated respiration was inhibited by 20 μM 3-BP within 10 min. Similar effects were also found in permeabilized hepatocytes of both species.

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Acknowledgments

This work was supported by grants PRVOUK P37/02 and SVV-2015-260179 from Charles University in Prague.

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Authors and Affiliations

  1. Department of Physiology, Faculty of Medicine in Hradec Kralove, Charles University in Prague, Hradec Kralove, Czech Republic

    Ondřej Sobotka, René Endlicher, Zdeněk Drahota, Otto Kučera, David Rychtrmoc, Marjan Raad, Khurum Hakeem & Zuzana Červinková

  2. Department of Anatomy, Faculty of Medicine in Hradec Kralove, Charles University in Prague, Hradec Kralove, Czech Republic

    René Endlicher

  3. Institute of Physiology, The Czech Academy of Sciences, Prague, Czech Republic

    Zdeněk Drahota

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  1. Ondřej Sobotka
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Correspondence to Zuzana Červinková.

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Sobotka, O., Endlicher, R., Drahota, Z. et al. Impaired mitochondrial functions contribute to 3-bromopyruvate toxicity in primary rat and mouse hepatocytes. J Bioenerg Biomembr 48, 363–373 (2016). https://doi.org/10.1007/s10863-016-9674-8

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