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Toxicity of Copper on Isolated Liver Mitochondria: Impairment at Complexes I, II, and IV Leads to Increased ROS Production

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Abstract

Oxidative damage has been implicated in disorders associated with abnormal copper metabolism and also Cu2+ overloading states. Besides, mitochondria are one of the most important targets for Cu2+, an essential redox transition metal, induced hepatotoxicity. In this study, we aimed to investigate the mitochondrial toxicity mechanisms on isolated rat liver mitochondria. Rat liver mitochondria in both in vivo and in vitro experiments were obtained by differential ultracentrifugation and the isolated liver mitochondria were then incubated with different concentrations of Cu2+. Our results showed that Cu2+ induced a concentration and time-dependent rise in mitochondrial ROS formation, lipid peroxidation, and mitochondrial membrane potential collapse before mitochondrial swelling ensued. Increased disturbance in oxidative phosphorylation was also shown by decreased ATP concentration and decreased ATP/ADP ratio in Cu2+-treated isolated mitochondria. In addition, collapse of mitochondrial membrane potential (MMP), mitochondrial swelling, and release of cytochrome c following of Cu2+ treatment were well inhibited by pretreatment of mitochondria with CsA and BHT. Our results showed that Cu2+ could interact with respiratory complexes (I, II, and IV). This suggests that Cu2+-induced liver toxicity is the result of metal’s disruptive effect on liver hepatocyte mitochondrial respiratory chain that is the obvious cause of Cu2+-induced ROS formation, lipid peroxidation, mitochondrial membrane potential decline, and cytochrome c expulsion which start cell death signaling.

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Abbreviations

ROS:

Reactive oxygen species

GSH:

Reduced glutathione

DCFH-DA:

2′,7′-dichlorofluorescin diacetate

TBARs:

Thiobarbituric acid reactive substances

Cs A:

Cyclosporin A

MDA:

Malondialdehyde

Rh123:

Rhodamine 123

BSA:

Bovine serum albumin

MPT:

Mitochondrial permeability transition

MMP:

Mitochondrial membrane potential

BHT:

Butylated hydroxytoluene

MTT:

3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide

TTFA:

Thenoyltrifuoroacetone

AA:

Antimycin A

DTNB:

Dithiobis-2-nitrobenzoic acid

Rot:

Rotenone

DMSO:

Dimethyl sulfoxide

TEP:

Tetramethoxypropane

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Acknowledgments

The investigation described in this paper was extracted from Ph.D. thesis of Dr. Mir-Jamal Hosseini (graduated from Faculty of Pharmacy, Shahid Beheshti University of Medical Sciences in 2012). The thesis conducted under supervision of Prof. Jalal Pourahmad at Faculty of Pharmacy, Shahid Beheshti University of Medical Sciences, Tehran, Iran. This study was supported by a grant from Shahid Beheshti University of Medical Sciences, Deputy of Research (88-01-94-6677, 2011).

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Authors and Affiliations

  1. Faculty of Pharmacy, Shahid Beheshti University of Medical Sciences, P. O. Box: 14155-6153, Tehran, Iran

    Mir-Jamal Hosseini, Fatemeh Shaki & Jalal Pourahmad

  2. Department of Pharmacology and Toxicology, School of Pharmacy, Zanjan University of Medical Sciences, Zanjan, Iran

    Mir-Jamal Hosseini

  3. Department of Pharmacology and Toxicology, School of Pharmacy, Mazandaran University of Medical Sciences, Sari, Iran

    Fatemeh Shaki

  4. Department of Pharmacology, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran

    Mahmoud Ghazi-Khansari

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  1. Mir-Jamal Hosseini
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  4. Jalal Pourahmad
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Correspondence to Jalal Pourahmad.

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Hosseini, MJ., Shaki, F., Ghazi-Khansari, M. et al. Toxicity of Copper on Isolated Liver Mitochondria: Impairment at Complexes I, II, and IV Leads to Increased ROS Production. Cell Biochem Biophys 70, 367–381 (2014). https://doi.org/10.1007/s12013-014-9922-7

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