153 ALLEVIATIVE EFFECTS OF ANTIOXIDANT ADMINISTRATION ON MATERNAL HYPERTHERMIA-INDUCED EARLY EMBRYONIC DEATH IN MICE

Hyperthermia-induced early embryonic death is generally ascribed to the high susceptibility of early embryos to elevated maternal body temperature. However, recent studies have indicated that the disruption of embryonic development by maternal hyperthermia has relevance to not only high temperature exposure to the embryo, but also hyperthermia-associated changes in the maternal body. Hyperthermia enhances physiological production of reactive oxygen species (ROS) systemically. Early embryos are susceptible to oxidative stress and it becomes easy to arrest their development when the oxidative stress is exposed. These findings led us to speculate that maternal hyperthermia-induced early embryonic death is caused by an increase in oxidative stress to the embryo. Vitamin E and melatonin are both well known to function as antioxidants in vivo when they are administered exogenously. In this study, we administered vitamin E or melatonin to heat-stressed pregnant mice, aiming to alleviate the hyperthermia-induced embryonic death. Female mice were heat-stressed (35�C with 60% relative humidity) for 12 h on the day of mating. In Experiment 1, vitamin E (1000 mg/kg body weight) was injected intraperitoneally just before stress treatment, or melatonin (3 mg/kg body weight) was injected subcutaneously for every 2 h during heat exposure. Then, zygotes were collected and in vitro developmental ability was assessed. In Experiment 2, intracellular glutathione (GSH) content in the zygote, ROS level and free radical scavenging activity (FRSA) in the oviduct, and lipid peroxidation level in the liver were measured to evaluate the effects of melatonin administration on physiological redox status. All data were analyzed using one-way ANOVA followed by Fisher's protected least-significant test. The percentage of embryos that developed to the morula or blastocyst stage was significantly (P < 0.01) increased by administration of either vitamin E (58.6%) or melatonin (47.9%) compared with that in heat-stressed mice that were administered placebos (14.8%). Intracellular GSH content in zygotes derived from melatonin-administered mice was not significantly different from that in unstressed mice (1.67 and 1.82 pmol/zygote, respectively), whereas in heat-stressed mice that were administered placebo, intracellular GSH content was significantly decreased (1.48 pmol/zygote, P < 0.05) compared to that in unstressed mice. There were no significant differences in lipid peroxidation levels in the liver and in ROS levels in the oviduct between melatonin-administered stressed mice and unstressed mice, although these parameters in heat-stressed mice with placebo were significantly higher than in unstressed mice (P < 0.05). Furthermore, FRSA in the oviduct was significantly (P < 0.05) higher in the melatonin-administered mice than in the heat-stressed mice. These findings suggest that antioxidant administration to heat-stressed mice alleviates the hyperthermia-induced early embryonic death, and this may be accomplished in part by maintaining a neutral redox status within the mother.