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Cytotoxic effects of hydroxylated fullerenes on isolated rat hepatocytes via mitochondrial dysfunction

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Abstract

The cytotoxic effects of hydroxylated fullerenes, also termed fullerenols or fullerols [C60(OH) n ], which are known nanomaterials and water-soluble fullerene derivatives, were studied in freshly isolated rat hepatocytes. The exposure of hepatocytes to C60(OH)24 caused not only concentration (0–0.25 mM)- and time (0–3 h)-dependent cell death accompanied by the formation of cell blebs, loss of cellular ATP, reduced glutathione (GSH), and protein thiol levels, but also the accumulation of glutathione disulfide and malondialdehyde, indicating lipid peroxidation. Of the other analogues examined, the cytotoxic effects of C60(OH)12 and fullerene C60 at a concentration of 0.125 mM were less than those of C60(OH)24. The loss of mitochondrial membrane potential and generation of oxygen radical species in hepatocytes incubated with C60(OH)24 were greater than those with C60(OH)12 and fullerene C60. In the oxygen consumption of mitochondria isolated from rat liver, the ratios of state-3/state-4 respiration were more markedly decreased by C60(OH)24 and C60(OH)12 compared with C60. In addition, C60(OH)24 and C60(OH)12 resulted in the induction of the mitochondrial permeability transition (MPT), and the effects of C60(OH)12 were less than those of C60(OH)24. Taken collectively, these results indicate that (a) mitochondria are target organelles for fullerenols, which elicit cytotoxicity through mitochondrial failure related to the induction of the MPT, mitochondrial depolarization, and inhibition of ATP synthesis in the early stage and subsequently oxidation of GSH and protein thiols, and lipid peroxidation through oxidative stress at a later stage; and (b) the toxic effects of fullerenols may depend on the number of hydroxyl groups participating in fullerene in rat hepatocytes.

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Abbreviations

DCHF-DA:

2′,7′-Dichlorodihydrofluorescein diacetate

DMSO:

Dimethyl sulfoxide

GSH:

Glutathione

GSSG:

Glutathione disulfide

HEPES:

N-(2-hydroxyethyl)-piperazine-N-(2-ethanesulfonic acid)

MDA:

Malondialdehyde

MPT:

Mitochondrial permeability transition

ROS:

Reactive oxygen species

ΔΨ:

Membrane potential

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The authors declare that they have no conflict of interest in this study.

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Authors and Affiliations

  1. Division of Pharmacology, Tokyo Metropolitan Institute of Public Health, 3-24-1, Hyakunin-cho, Shinjuku-ku, Tokyo, 169-0073, Japan

    Yoshio Nakagawa, Dai Nakae & Akio Ogata

  2. Division of Water Environment, Tokyo Metropolitan Institute of Public Health, 3-24-1, Hyakunin-cho, Shinjuku-ku, Tokyo, 169-0073, Japan

    Toshinari Suzuki

  3. Department of Molecular and Cellular Pathophysiology, Showa Pharmaceutical University, 3-3365, Higashi Tamagawa Gakuen, Machida, Tokyo, 194-8543, Japan

    Hidemi Ishii

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  1. Yoshio Nakagawa
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  2. Toshinari Suzuki
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Correspondence to Yoshio Nakagawa.

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Nakagawa, Y., Suzuki, T., Ishii, H. et al. Cytotoxic effects of hydroxylated fullerenes on isolated rat hepatocytes via mitochondrial dysfunction. Arch Toxicol 85, 1429–1440 (2011). https://doi.org/10.1007/s00204-011-0688-z

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  • DOI: https://doi.org/10.1007/s00204-011-0688-z

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