| Summary: | 碩士 === 臺北醫學大學 === 醫學技術學系 === 93 === Nonylphenol (NP) is a degradation product of a widely used nonionic surfactant nonylphenol ethoxylate (NPEO). NP has been identified as an environmental hormone with estrogenic activity, and suggested to disrupt normal reproductive function of mammals. In this study, we investigated the causal roles of NP in inducing trophoblast cell death and impaired ICR mouse embryo development. We conducted several experiments to address the NP-induced effects on cellular viability, mitochondrial function, and identified the molecular mechanism of cytotoxicity in NP-treated human trophoblast cells (3A-Sub-E). The results showed that the cell viability was declined to 51.54±16.65% in 1.5 uM NP-treated cells by dye exclusion assay. Reactive oxygen species (ROS) generation and oxidative damages were increased in a dose- and time-dependent manner. The dose-dependent decrease of ATP production and mtDNA copy number in NP-treated cells were detected by luminescence assay and real-time quantitative PCR. Furthermore, we found the hypoxia inducible factor-1a (HIF-1a) was increased by NP via ROS generation. The increased inducible nitric oxide synthase (iNOS) expression and nitric oxide (NO) generation were also detected in NP-treated cells. The iNOS protein was increased 58.00±7.94% and NO generation was measured 23.27±6.91% in the 1 uM NP-treated cells. After 2 uM NP treatment, approximate 46.47±0.26% cells underwent necrosis and 22.05±1.61% cells were apoptotic when detected by Annexin V and PI staining. In addition, we also demonstrated the detrimental effects of NP on embryonic development capacity. The increased hatching rate were found in 4-cell embryos with 1 uM NP treatment. However, the increased abnormal morphology was found in the hatched embryos with NP treatment. In conclusion, that NP contributes to the induction of oxidative damages in placenta, and may affect embryo viability and developmental competence.
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