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Ex vivo Activities of β‐Lapachone and α‐Lapachone on Macrophages: A Quantitative Pharmacological Analysis Based on Amperometric Monitoring of Oxidative Bursts by Single Cells
Author(s) -
Ferreira Danielle C. M.,
Tapsoba Issa,
Arbault Stéphane,
Bouret Yann,
Alexandre Moreira Magna Suzana,
Ventura Pinto Antônio,
Goulart Marília O. F.,
Amatore Christian
Publication year - 2009
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.200800517
Subject(s) - ex vivo , amperometry , chemistry , oxidative phosphorylation , in vivo , oxidative stress , biochemistry , microbiology and biotechnology , biology , in vitro , electrode , electrochemistry
Artificial synapses for femtomolar detection : Amperometry at platinized carbon fibre electrodes has been used to unravel the complexity of β‐lapachone's effects on cellular oxidative stress. α‐Lapachone, the pharmacologically inactive para ‐quinone isomer, did not display such characteristics, but over longer incubation periods both quinones induced apoptosis. The observed effects were interpreted in terms of two mechanisms involving opposite reactivities of quinones in living cells.β‐Lapachone ( 1 ) has been widely used for its pharmacological activity, particularly against cancer. However, its mechanism of action at the cellular level remains unclear, although a common major hypothesis involves its prooxidant properties. Electrochemical measurements with microelectrodes were taken in order to quantitatively investigate the activity of 1 at different concentrations and several incubation times, on the oxidative bursts released by single macrophages. The exact natures of the electroactive reactive oxygen species (ROS) and reactive nitrogen species (RNS) released by macrophages under the effect of 1 were characterized, and their fluxes were measured quantitatively. This allowed the reconstruction of the primary O 2 .− and NO production by the cells. In the first hour, at 10 μ M , the decrease in the oxidative burst involved mainly RNS, while the amount of H 2 O 2 was found to be higher than in controls. After a longer incubation time—that is, 4 h—at 1 μ M , the total amount of ROS and RNS had increased, with significant enhancements of H 2 O 2 and NO. In contrast, α‐lapachone, the pharmacologically inactive para ‐quinone isomer, was unable to increase the production of RONS by macrophages significantly. Over much longer incubation periods (about one day), however, each quinone induced cell death by apoptosis. All these effects were interpreted by consideration of two different mechanisms involving opposite reactivities of quinones in living cells.