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Methyl tert ‐butyl ether (MTBE)‐induced cytotoxicity and oxidative stress in isolated rat spermatogenic cells
Author(s) -
Li Dongmei,
Yin Daqiang,
Han Xiaodong
Publication year - 2006
Publication title -
journal of applied toxicology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.784
H-Index - 87
eISSN - 1099-1263
pISSN - 0260-437X
DOI - 10.1002/jat.1178
Subject(s) - dichlorofluorescein , oxidative stress , propidium iodide , chemistry , lipid peroxidation , superoxide dismutase , reactive oxygen species , biochemistry , microbiology and biotechnology , apoptosis , biology , programmed cell death
Methyl tert ‐butyl ether (MTBE) is a class of synthetic organic chemical. In the USA, MTBE pollution is regarded as a serious environmental problem. The objective of the present study was to investigate the cytotoxic effects and oxidative stress induced by MTBE in isolated rat spermatogenic cells. In cytotoxic experiments, spermatogenic cells isolated from the testes of adult Sprague‐Dawley rats by a mechanical procedure without the use of trypsin were incubated with medium alone (control), 0.5, 5, 50 m m MTBE, respectively, for 6, 12 and 18 h. MTT assay, staining with fluorescein diacetate (FDA) and propidium iodide (PI) and flow cytometric analyses were used. In oxidative stress experiments, the spermatogenic cells were incubated with medium alone (control) and with 0.5, 50 μm, 5 m m MTBE. For 1, 2, 6, 12, 18 h incubation, ROS production was tested using a 2′,7′‐dichlorofluorescein diacetate (DCHF‐DA) probe; for 1, 3, 6, 12, 18 h incubation, cytosolic superoxide dismutase (SOD) and extracellular SOD (SOD EX ) activity was assessed; and for 18 h incubation, lipid peroxidation was assessed. The results showed that MTBE at high doses significantly decreased the spermatogenic cell viability and increased plasma membrane damage and the ratio of necrotic cells compared with the control. Assessment of the MTBE‐induced oxidative stress revealed that MTBE increased the production of reactive oxygen species (ROS) and enhanced lipid peroxidation. In addition, although SOD EX activity increased at a high dose level, cytosolic SOD activity decreased. These results suggest that an increase of MTBE‐induced ROS production and an enhancement of membrane lipid peroxidation may play an important role in its cytotoxicity in isolated rat spermatogenic cells. Copyright © 2006 John Wiley & Sons, Ltd.