T-2 toxin exposure induces apoptosis in rat ovarian granulosa cells through oxidative stress
Jing Wu1, Di Tu1, Li-Yun Yuan1
1College of Veterinary Medicine, Hunan Agricultural University, Changsha, Hunan 410128, PR China.
Objective:
To investigate the reproductive toxicity and cytotoxicity of T-2 toxin, which is a mycotoxin, and to explore its potential apoptotic induction mechanism.
Methods:
ovarian granulosa cells of rats were treated with T-2 toxin (1-100nM) for 24h. The cytotoxicity was assessed with MTT bioassay and apoptotic cells were detected by flow cytometry, and further identified by chromatin condensation and nuclear fragmentation with Hoechst 33258 under microscope; reactive oxygen species (ROS) with DCFH-DA was analyzed by fluorometry; total superoxide dismutase (SOD) was determined by NBT staining method. Glutathione peroxidase (GSHPx) activity was detected by UV colorimetric assay. The activity of catalase (CAT) in granulosa cells was measured by the Aebi method, and malondialdehyde (MDA) content was determined by thiobarbituric acid assay.
Results:
T-2 toxin dose-dependently inhibited the growth of granulosa cells and resulted in apoptosis in rat granulosa cells. Treatment with T-2 toxin could induce ROS and MDA accumulation in granulosa cells, acompanying with losses of activities of SOD, GSH-Px and CAT, whereas T-2 toxin-induced apoptosis in granulosa cells could be significantly inhibited through the use of antioxidant Trolox.
Conclusion:
These data suggest that T-2 toxin may induce the apoptosis in rat granulosa cells through oxidative stress.
