期刊
FREE RADICAL BIOLOGY AND MEDICINE
卷 81, 期 -, 页码 69-76出版社
ELSEVIER SCIENCE INC
DOI: 10.1016/j.freeradbiomed.2015.01.015
关键词
Hydrogen peroxide; Reactive oxygen species; Human spermatozoa; Organic synthesis; Aryl boronate; Free radicals
资金
- Cook Medical Pty Ltd
- Australian Research Council Linkage Grant [LP 110200736]
- ARC Centre of Excellence in Nanoscale BioPhotonics
- ARC Georgina Sweet Laureate Fellowship [FL130100044]
- [DP 110103951]
Human spermatozoa are compromised by production of reactive oxygen species (ROS), and detection of ROS in spermatozoa is important for the diagnosis of male infertility. The probes 2',7'-dichlorohydrofluorescein diacetate (DCFH), dihydroethidium (DHE), and MitoSOX red (MSR) are commonly used for detecting ROS by flow cytometry; however, these probes lack sensitivity to hydrogen peroxide (H2O2), which is particularly damaging to mammalian sperm cells. This study reports the synthesis and use of three aryl boronate probes, peroxyfluor-1 (PF1), carboxyperoxyfluor-1, and a novel probe, 2-(2-ethoxyethoxy)ethoxyperoxyfluor-1 (EEPF1), in human spermatozoa. PF1 and EEPF1 were effective at detecting H2O2 and peroxynitrite (ONOO-) produced by spermatozoa when stimulated with menadione or 4-hydroxynonenal. EEPF1 was more effective at detection of ROS in spermatozoa than DCFH, DHE, or MSR; furthermore it distinguished poorly motile sperm as shown by greater ROS production. EEPF1 should therefore have a significant role in the diagnosis of oxidative stress in male infertility, cryopreservation, age, lifestyle, and exposure to environmental toxicants. (C) 2015 Elsevier Inc. All rights reserved.
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