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Protective effect of safranine on the mitochondrial damage induced by Fe(II)citrate: comparative study with trifluoperazine

  • R. F. Castilho
  • R. S. Pereira
  • A. E. Vercesi
Article

Summary

In this study, we show that safranine at the concentrations usually employed as a probe of mitochondrial membrane potential significantly protects against the oxidative damage of mitochondria induced by Fe(II)citrate. The effect of safranine was illustrated by experiments showing that this dye strongly inhibits both production of thiobarbituric acid-reactive substances and membrane potential decrease when energized mitochondria were exposed to Fe(II)citrate in the presence of Ca2+ ions. Similar results were obtained with the lipophylic compound trifluoperazine. It is proposed that, like trifluoperazine, safranine decreases the rate of lipid peroxidation due to its insertion in the membrane altering the physical state of the lipid phase.

Keywords

Safranine mitochondrial damage Fe(II)citrate trifluoperazine 

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References

  1. 1.
    Dell’Antone P., Colonna R., Azzone G.F. (1972): The membrane structure studied with cationic dyes. I. The binding of cationic dyes to submitochondrial particles and the question of the polarity of the ion-translocation. Eur. J. Biochem., 24, 553–565.CrossRefPubMedGoogle Scholar
  2. 2.
    Colonna R., Massari S., Azzone G.F. (1973): The problem of cation-binding sites in the energized membrane of intact mitochondrial. Eur. J. Biochem., 34, 577–585.CrossRefGoogle Scholar
  3. 3.
    Åkerman K.E.O., Wikström M.K.F. (1976): Safranine as a probe of the mitochondrial membrane potential, FEBS Lett., 68, 191–197.CrossRefPubMedGoogle Scholar
  4. 4.
    Zanotti A., Azzone G.F. (1980): Safranine as membrane potential probe in rat liver mitochondria. Arch. Biochem. Biophys., 201, 255–265.CrossRefPubMedGoogle Scholar
  5. 5.
    Valle V.G.R., Pereira-da-Silva L., Vercesi A.E. (1986): Undesirable feature of safranine use as a probe for mitochondrial membrane potential. Biochem. Biophys. Res. Commn., 135, 189–195.CrossRefGoogle Scholar
  6. 6.
    Schneider W. C., Hogerboom G. H. (1950): Intracellular distribution of enzymes. V. Further studies on the distribution of cytochrome a in rat liver homogenates. J. Biol. Chem. 183, 213–228.Google Scholar
  7. 7.
    Kamo N., Muratsugo M., Ruji H., Kobatake J. (1979): Membrane potential of mitochondria measured with an electrode sensitive to tetraphenyl phosphonium and relationship between proton electrochemical potential phosphorylation potential in steady state. J. Membr. Biol., 48, 105–121.Google Scholar
  8. 8.
    Muratsugu M., Kamo N., Kurihara K., Kobatake J. (1977): Selective electrode for dibenzyl ammonium cation as indicator of the membrane potential in biological systems. Biochim. Biophys. Acta, 464, 613–619.CrossRefPubMedGoogle Scholar
  9. 9.
    Jensen B.D., Gunter K.K., Gunter T.E. (1986): The efficiencies of the component steps of oxidative phosphorylation. II. Experimental determination of the efficiencies in mitochondria and examination of the equivalence of membrane potential and pH gradient in phosphorylation. Arch. Biochem. Biophys. 248, 305–323.CrossRefPubMedGoogle Scholar
  10. 10.
    Vercesi A.E., Bernardes C.F., Hoffmann M.E., Gadelha F.R., Docampo R. (1991): Digitonin permeabilization does not affect mitochondrial function and allows the determination of the mitochondrial membrane potential ofTrypanosoma cruzi in situ. J. Biol. Chem., 266, 14431–14434.PubMedGoogle Scholar
  11. 11.
    Buege J.A., Aust S.D. (1978): Microsomal lipid peroxidation. Methods Enzymol., 52, 302–310.CrossRefPubMedGoogle Scholar
  12. 12.
    Castilho R.F., Meinicke A.R., Almeida A.M., Hermes-Lima M., Vercesi A.E. (1994): Oxidative damage of mitochondria induced by Fe(II)citrate is potentiated by Ca2+ and includes lipid peroxidation and alterations in membrane proteins. Arch. Biochem. Biophys., 308, 158–163.CrossRefPubMedGoogle Scholar
  13. 13.
    Castilho R.F., Kowaltowski A.J., Meinicke A.R., Vercesi A.E. (1995): Oxidative damage of mitochondria induced by Fe(II)citrate ort-butyl hydroperoxide in the presence of Ca2+: effect of coenzyme Q redox state. Free Rad. Biol. Med., 18, 55–59.CrossRefPubMedGoogle Scholar
  14. 14.
    Slater T.F. (1968): The inhibitory effects ‘in vitro’ of phenothiazines and other drugs on lipid peroxidation systems in rat liver microsomes and their relatioship to liver necrosis produced by carbon tetrachloride. Biochem. J., 106, 155–160.PubMedGoogle Scholar
  15. 15.
    Janero D.R., Burghardt B. (1989): Prevention of oxidative injury to cardiac phospholipids by membrane-active ‘stabilizing agents’. Res. Commun. Chem. Pathol. Pharmacol., 63, 163.PubMedGoogle Scholar
  16. 16.
    Breugnot C., Mazière C., Salmon S., et al. (1990): Phenothiazines inhibit copper and endothelial cell-induced peroxidation of low density lipoprotein. Biochem. Pharmacol., 40, 1975–1980.CrossRefPubMedGoogle Scholar
  17. 17.
    Pereira R.S., Bertocchi A.P.F., Vercesi A.E. (1992): Protective effect of trifluoperazine on the mitochondrial damage induced by Ca2+ plus prooxidants. Biochem. Pharmacol., 44, 1795–1801.CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag 1996

Authors and Affiliations

  • R. F. Castilho
    • 1
  • R. S. Pereira
    • 2
  • A. E. Vercesi
    • 1
  1. 1.Departamento de Bioquímica, IBUNICAMPSão PauloBrazil
  2. 2.Departamento de Bioquímica, IQUNESPAraraquara. SPBrazil

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