The Effects and Molecular Mechanisms of Phthalates (DEHP 和DBP) on the Mitochondrial Function and Genetic Regulation in Ovarian Granulosa Cells and Mouse Embryo

Endocrine disrupting compounds (EDCs), including phthalates (PAEs), are chemicals that interfere with human hormonal system and cause cancerous tumors, birth defects, and other developmental disorders. PAEs are a group of related compounds that are very widely used as plasticizers and solvents. However, effects of PAEs on reproductive system are still obscures. In this 3-year study, we are going to investigate the reproductive effects of Di-(2-ethylhexyl) phthalate (DEHP) and Di-n-butyl phthalate (DBP) on woman reproductive system by evaluation of these compounds on ovarian granulosa cells and mouse embryo. In our preliminary experimental results, we had already found that high concentration of DEHP (10 and 100 μM) exposure in mouse could reduce the number of ovarian granulosa cells. Moreover, administration with DEHP for 2 weeks increased intracellular reactive oxygen species (ROS) and mitochondrial activity. There are at least three major and specific aims will be accomplished in this research: (1) To evaluate the following effects of PAEs (DEHP and DBP) on ovarian granulosa cells, including median lethal dose (LD50), impacts on secretion of estradiol, progesterone and Anti-Müllerian hormone (AMH) levels, intracellular calcium concentration, lipid peroxidation evaluation and mitochondrial copy number, morphology and activity. (2) To determine the effects of PAEs on telomerase activity in mouse embryo by analysis of related genes and proteins expression profiles (leukemia inhibitory factor receptor, hypoxia inducible factor-1 alpha, hypoxia inducible factor-2 alpha, manganese superoxide dismutase and vascular endothelial growth factor, etc.) (3) Comparison of gene expression profile by transcriptome and microRNA analysis. The interference of phthalates and the metabolites on epigenetic regulation in mouse embryo and fetal development will also be examined. Through this project, it is helpful to clarify the effects and the action mechanisms of PAEs on the reproductive systems. It may provide the therapeutic strategies to target the complex pathway in embryonic development and further fertility protection.