2-Phenyl-β-lapachone can affect mitochondrial function by redox cycling mediated oxidation

doi:10.1016/j.abb.2004.09.020

Archives of Biochemistry and Biophysics

Volume 432, Issue 2, 15 December 2004, Pages 129-135

https://doi.org/10.1016/j.abb.2004.09.020 Get rights and content

Abstract

2-Phenyl-β-lapachone (3,4-dihydro-2-methyl-2-phenyl-2H-naphtho[1,2b]pyran-5,6-dione) (2PBL) is a o-naphthoquinone synthesized as a possible antitumoral agent. The addition of micromolar concentrations of 2PBL to rat liver mitochondria (in the presence of malate-glutamate or succinate, as respiratory substrates): (1) stimulated O₂ consumption in state 4 and inhibited O₂ consumption in state 3, thus decreasing respiratory control index (RCI); and (2) collapsed the mitochondrial membrane potential. The addition of 2PBL to rat liver submitochondrial particles: (1) stimulated NADH oxidation in the presence of rotenone, antimycin, myxothiazol or cyanide; (2) stimulated $^{} O_{2}^{-}$ production in the presence of NADH and antimycin; and (3) led to 2PBL semiquinone radical production. Control studies carried out with two p-naphthoquinones, menadione and atovaquone, did not produced equivalent effects. These findings support the hypothesis that 2PBL, undergoes redox cycling and affects mitochondrial function. The 2PBL effect is complex, involving inhibition of electron transfer, uncoupling of oxidative phosphorylation, collapse of mitochondrial membrane potential and $^{} O_{2}^{-}$ production by redox cycling. The mitochondrion could be a target organelle for 2PBL cytotoxicity.

Section snippets

Chemicals

2PBL and 2-ethyl-β-lapachone were obtained from Novartis (Ciba-Geigy), Basle, Switzerland. Atovaquone was provided by Welcome Laboratories, Langley Court, Beckenham, Kent, UK. Naphthoquinones were used dissolved in dimethylformamide (DMFA). KCl, KCN, and MgCl₂ were purchased from Mallinckrodt Chemical Works, New York, USA. DMFA, bovine serum albumin (A-4503), ADP, l-glutamate, l-malate, l-malonate, l-succinate, NADH, sucrose, Tris(hydroxymethyl)aminomethane, menadione, rotenone, antimycin A,

Effect of 2PBL, menadione and atovaquone, on electron transfer and oxidative phosphorylation in coupled mitochondria

Table 1 shows the effect of 2PBL, menadione and atovaquone on the respiration rate of coupled mitochondria. Using malate–glutamate as substrate, 2PBL inhibited state 3 mitochondrial respiration at 10 μM, increased state 4 respiration at 5 μM and decreased RCI at 2.5 μM. Using succinate as substrate, 2PBL inhibited state 3 mitochondrial respiration at 25 μM, activated state 4 at 5 μM and decreased RCI at 2.5 μM. Similar results were obtained with 2-ethyl-β-lapachone (data not shown).

Equivalent

Discussion

Our results show that, within the 2.5–50 μM range, 2PBL induces changes in mitochondrial respiration states, where there is an increased rate of $^{} O_{2}^{-}$ production, an inhibition of mitochondrial electron transport, the uncoupling of oxidative phosphorylation and the collapse of mitochondrial membrane potential.

The NADH oxidation, insensitive to rotenone, antimycin, myxothiazol or cyanide (Fig. 2) and the significant correlation between 2PBL effect on NADH oxidation and $^{} O_{2}^{-}$ production in

Acknowledgments

This work was aided in part by grants from CONICET and Fundación Navarro Viola (Dr. C. Valiente Noailles), Buenos Aires, Argentina. Dr. Natacha de Witte was a fellow from Academy of Sciences, Buenos Aires (Dr. Rodolfo Brenner). We thank Dr. Alicia Fuchs for helpful discussion and Mrs. A. Verón, S. del Valle and C. Lincon for technical assistance.

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^✠: Professor Stoppani passed away on March 18, 2003.

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2-Phenyl-β-lapachone can affect mitochondrial function by redox cycling mediated oxidation

Abstract

Section snippets

Chemicals

Effect of 2PBL, menadione and atovaquone, on electron transfer and oxidative phosphorylation in coupled mitochondria

Discussion

Acknowledgments

Biochem. Pharmacol.

Trends Comp. Biochem. Physiol.

Curr. Cancer Drug Targets

Biochem. Pharmacol.

Free Radic. Biol. Med.

Cancer Res.

Cancer Res.

Cancer Res.

Oncol. Rep.

Exp. Eye Res.

Mol. Med.

Mol. Med.

J. Med. Chem.

Anticancer Drugs

Proc. Natl. Acad. Sci. USA

J. Biol. Chem.

Biochem. J.

Methods Enzymol.