Effect And Mechanisms Of Plasticezer Dehp On Embryo Implantation

Plasticizer DEHP is a ubiquitous environmental pollutant with significant human exposure. It causes serious adverse effects on animal and human health, especially on reproductive system, i.e., leads to retained testicle and interrupted spermatogenesis in males. In fact, women are exposed to DEHP more frequently than men due to its wide use in cosmetics and shampoos. Current data has indicated that exposure of females to DEHP was associated with harmful pregnancy outcomes, but the mechanism is still unclear. Embryo implantation, as an important step of pregnancy, is influenced by many factors. One key factor is endometrial receptivity, it regulated by estrogen and/or progesterone which act primarily through their respective nuclear receptors and downstream effectors. Moderate invasion of extravillous trophoblast cells is another key factor for embryo implantation. Whether they were affected by DEHP and lead to failure of pregnancy are worth to investigate.To investigate the effects of DEHP embryo implantation and its molecular mechanism, the follow experiments were done. First, we use mouse as research object to investigate the effect of DEHP on endometrial receptivity and embryo implantation. The main contents include:The mated mice received DEHP from D1until sacrifice, the collected uteri and counted the number of implanted sites; followed check the morphological of uteri associated with endometrial receptivity using H&E and SEM; And Real-time PCR, western blot and Immunohistochemistry were used to detected the expression alternation of ERa, PR, E-cadherin, HoxA10, MMP-2and their signaling pathway associated with endometrial receptivity. Secondly, we detected the effect and mechanism of DEHP on invasion and migration of human extravillous trophoblast using HTR-8/Svneo cells. The concert experiments as follow:we fist used MTT assay and flow cytometry to determine the optimum concentration of DEHP treating to HTR-8/Svneo cells; then detected the cell invasion and migration by Transwell chambers; lastly, the expression level of mRNA and protein which related to cell invasion and migration were detected by Real-time PCR and Western blot. The main results are as follows:1. DEHP led to compromised endometrial receptivity and decreased number of implantation sites, and produced harmful pregnancy outcome.2. The mRNA and protein expression of ERa, PR and E-cadherin, but not HoxA10and MMP-2, were up-regulated by DEHP in the mouse endometrium.3. DEHP altered E-cadherin levels in the mouse endometrium via the MAPK and NF-κB signaling pathways.4. DEHP enhanced HTR-8/Svneo cells invasion and migration5. DEHP up-regulated MMP-2and MMP-9expression through the phosphorylated of p38and JNK.Together all, DEHP reduces the number of implanted embryo through disrupting the endometrial receptivity of pregnancy mouse and invasion of HTR-8/Svneo cells, then produced harmful pregnancy outcome.