Supra-physiological doses of testosterone affect membrane oxidation of human neutrophils monitored by the fluorescent probe C11-BODIPY58¹/59¹.
Eur J Appl Physiol
; 113(5): 1241-8, 2013 May.
Article
in En
| MEDLINE
| ID: mdl-23160653
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-BODIPY(581/591). 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-BODIPY(581/591) 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, 10(2) to 10(4)-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-BODIPY(581/591) for monitoring (auto-induced) membrane oxidation as an important parameter of viability and, thus, responsiveness of immune cells in inflammatory processes.
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Testosterone
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Cell Membrane
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Oxidative Stress
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Neutrophils
Limits:
Adult
/
Humans
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Male
Language:
En
Journal:
Eur J Appl Physiol
Journal subject:
FISIOLOGIA
Year:
2013
Type:
Article
Affiliation country:
Brazil