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Evaluation of Mitochondrial Respiration in Cultured Rat Hepatocytes

  • Jean-Pierre Marchandeau
  • Gilles LabbeEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1641)

Abstract

Mitochondrial dysfunction is a major mechanism whereby drugs can induce liver injury and other serious side effects, such as lactic acidosis and rhabdomyolysis, in some patients. Several in vitro and in vivo investigations can be performed in order to determine if drugs can disturb mitochondrial fatty acid oxidation (FAO) and the oxidative phosphorylation (OXPHOS) process, deplete hepatic mitochondrial DNA (mtDNA), or trigger the opening of the mitochondrial permeability transition pore (MPT). Among these investigations, mitochondrial respiration is a relatively easy test to measure the potential toxicity of a drug. The use of cells instead of isolated mitochondria allows one to test the toxic effect of a parent compound and its metabolites. The use of rat hepatocytes can detect drugs involved in drug-induced liver injuries (DILI). The method consists in measuring oxygen consumption by using a Clark electrode in a chamber containing a suspension of hepatocytes preincubated with drug.

Key words

Mitochondrial respiration Oxygen consumption Clark electrode Respirometer Primary rat hepatocytes 

Notes

Acknowledgments

The authors would like to thank Delphine Hoët for her expertise and her support in hepatocyte culture.

References

  1. 1.
    Lee WM (2003) Drug-induced hepatotoxicity. N Engl J Med 349:474–485CrossRefPubMedGoogle Scholar
  2. 2.
    Pessayre D, Fromenty B, Mansouri A, Berson A (2007) Hepatotoxicity due to mitochondrial injury. In: Kaplowitz N, DeLeve LD (eds) Drug-induced liver disease, 2nd edn. Informa Healthcare, New York, USA, pp 49–84Google Scholar
  3. 3.
    Fromenty B, Pessayre D (1995) Inhibition of mitochondrial beta-oxidation as a mechanism of hepatotoxicity. Pharmacol Ther 67:101–154CrossRefPubMedGoogle Scholar
  4. 4.
    Pessayre D, Mansouri A, Haouzi D, Fromenty B (1999) Hepatotoxicity due to mitochondrial dysfunction. Cell Biol Toxicol 15:367–373CrossRefPubMedGoogle Scholar
  5. 5.
    Igoudjil A, Begriche K, Pessayre D, Fromenty B (2006) Mitochondrial, metabolic and genotoxic effects of antiretroviral nucleoside reverse-transcriptase inhibitors. Curr Med Chem: Anti-Infect Agents 5:273–292Google Scholar
  6. 6.
    Dykens JA, Will Y (2007) The significance of mitochondrial toxicity testing in drug development. Drug Discov Today 12:777–785CrossRefPubMedGoogle Scholar
  7. 7.
    Scatena R, Bottoni P, Botta G, Martorana GE, Giardina B (2007) The role of mitochondria in pharmacotoxicology: a reevaluation of an old, newly emerging topic. Am J Physiol Cell Physiol 293:C12–C21CrossRefPubMedGoogle Scholar
  8. 8.
    Salway JG (1994) Metabolism at a glance, 1st edn. Blackwell Scientific Publications, London, UKGoogle Scholar
  9. 9.
    Pessayre D, Fromenty B (2005) NASH: a mitochondrial disease. J Hepatol 42:928–940CrossRefPubMedGoogle Scholar
  10. 10.
    Navarro A, Boveris A (2007) The mitochondrial energy transduction system and the aging process. Am J Physiol Cell Physiol 292:C670–C686CrossRefPubMedGoogle Scholar
  11. 11.
    Berson A, Renault S, Letteron P, Robin MA, Fromenty B, Fau D, Le Bot MA, Riche C, Durand-Schneider AM, Feldmann G, Pessayre D (1996) Uncoupling of rat and human mitochondria: a possible explanation for tacrine-induced liver dysfunction. Gastroenterology 110:1878–1890CrossRefPubMedGoogle Scholar
  12. 12.
    Korzeniewski C, Callewaert DM (1983) An enzyme-release assay for natural cytotoxicity. J Immunol Methods 64:313–320CrossRefPubMedGoogle Scholar
  13. 13.
    Crouch SPM, Kozlowski R, Slater KJ, Fletcher J (1993) The use of ATP bioluminescence as a measure of cell proliferation and cytotoxicity. J Immunol Methods 160:81–88CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  1. 1.Investigative Toxicology, Preclinical SafetySanofi R&DVitry-sur-SeineFrance

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