Supra-physiological doses of testosterone affect membrane oxidation of human neutrophils monitored by the fluorescent probe C11-BODIPY581/591

Authors

Tacito Pessoa de Souza Jr, Paraná Federal University
Andre K. Yamada, São Carlos Federal University
Roberto Simao, Federal University of Rio de Janeiro
Tatiana G. Polotow, Institute of Physical Activity and Sports Sciences (ICAFE), Universidade Cruzeiro do Sul
Rui Curi, University of Sao Paulo
Zachary Pope, Eastern Illinois University
Jeffrey Willardson, Eastern Illinois UniversityFollow
Marcelo P. Barros, Institute of Physical Activity and Sports Sciences (ICAFE), Universidade Cruzeiro do Sul

Document Type

Article

Publication Date

May 2013

Abstract

The purpose of this study was to determine the effects of supra-physiological doses of testosterone (TES) on membrane oxidation of activated human neutrophils in vitro using an innovative and sensitive technique: the real-time detection with the fluorescence probe C11-BODIPY581/591. Methodological controls were performed with the lipid-soluble and powerful antioxidant astaxanthin at different neutrophil density cultures. Neutrophils from nine healthy young men (23.4 ± 2.5 years, 174.4 ± 7.0 cm height, and 78.3 ± 7.0 kg weight) were isolated and treated with 0.1 or 10 μM TES for 24 h and subsequently labeled with the free radical-sensitive probe C11-BODIPY581/591 for monitoring membrane oxidation after neutrophil activation with phorbol-12-myristate-13-acetate (PMA). First-order exponential decay kinetic indicated that both 0.1 and 10 μM TES severely increased baseline membrane oxidation in non-activated human neutrophils (compared to control). However, similar kinetics of membrane oxidation were observed in control and 0.1 μM TES-treated neutrophils after PMA activation, whereas chemical activation did not alter the baseline higher rates of membrane oxidation in 10 μM TES-treated neutrophils. The data presented here support the hypothesis that TES exerts distinct effects on the membrane oxidation of human neutrophils, depending on its dose (here, 102 to 104-fold higher than physiological levels in men) and on PMA activation of the oxidative burst. Furthermore, this paper also presents an innovative application of the free radical-sensitive probe C11-BODIPY581/591 for monitoring (auto-induced) membrane oxidation as an important parameter of viability and, thus, responsiveness of immune cells in inflammatory processes.

Comments

Final version available at http://link.springer.com/article/10.1007/s00421-012-2538-y/fulltext.html

Recommended Citation

de Souza, Tacito Pessoa Jr; Yamada, Andre K.; Simao, Roberto; Polotow, Tatiana G.; Curi, Rui; Pope, Zachary; Willardson, Jeffrey; and Barros, Marcelo P., "Supra-physiological doses of testosterone affect membrane oxidation of human neutrophils monitored by the fluorescent probe C11-BODIPY581/591" (2013). Faculty Research and Creative Activity. 29.
https://thekeep.eiu.edu/kss_fac/29