European Journal of Applied Physiology

, Volume 113, Issue 5, pp 1241–1248

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

  • Tácito Pessoa de Souza-Junior
  • André K. Yamada
  • Roberto Simão
  • Tatiana G. Polotow
  • Rui Curi
  • Zachary Pope
  • Jeffrey M. Willardson
  • Marcelo P. Barros
Original Article
  • 281 Downloads

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.

Keywords

Lipoperoxidation Steroid Astaxanthin Antioxidant Immune Oxidative stress 

Abbreviations

AAS

Androgen-anabolic steroids

AST

Astaxanthin

C11-BODIPY581/591

4,4-Difluoro-5-(4-phenyl-1,3-butadienyl)-4-bora-3a,4a-diaza-s-indacene-3-undecanoic acid

DMSO

Dimethylsulfoxide

HEPES

4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid

PMA

Phorbol-12-myristate-13-acetate

ROS/RNS

Reactive oxygen/nitrogen species

TBARS

Thiobarbituric acid reactive substances

TES

Testosterone

VSMCs

Vascular smooth muscle cells

Copyright information

© Springer-Verlag Berlin Heidelberg 2012

Authors and Affiliations

  • Tácito Pessoa de Souza-Junior
    • 1
  • André K. Yamada
    • 2
  • Roberto Simão
    • 3
  • Tatiana G. Polotow
    • 4
  • Rui Curi
    • 5
  • Zachary Pope
    • 6
  • Jeffrey M. Willardson
    • 6
  • Marcelo P. Barros
    • 4
  1. 1.Department of Physical EducationParaná Federal University (UFPR)CuritibaBrazil
  2. 2.Department of Physiological SciencesSão Carlos Federal UniversitySão CarlosBrazil
  3. 3.Federal University of Rio de Janeiro (UFRJ)Rio de JaneiroBrazil
  4. 4.Institute of Physical Activity and Sports Sciences (ICAFE)Universidade Cruzeiro do SulSão PauloBrazil
  5. 5.Department of Physiology and Biophysics, Biomedical Building IUniversity of Sao Paulo (USP-SP)Sao PauloBrazil
  6. 6.Department of Kinesiology and Sports StudiesEastern Illinois UniversityCharlestonUSA

Personalised recommendations