Di-2-ethylhexylphthalate-induced miR155-5P promotes placental ferroptosis

Growing attention has focused on DEHP's impact on women's early life stages, particularly its reproductive toxicity linked to oxidative stress and ferroptosis. This study explored DEHP's effect on female reproduction and its relationship with miR155-5P and ferroptosis. To address this, experiments were conducted using HTR-8/Svneo trophoblast cells and pregnant rat models. Key methods included CCK8, Western blot, ELISA, flow cytometry, qPCR, and dual-luciferase assays to assess cell proliferation, ferroptosis markers, and molecular interactions. Additionally, placental tissue changes of pregnant rats were analyzed by HE staining and molecular detection. Consistent with the experimental design, results demonstrated that DEHP suppressed cell proliferation and induced ferroptosis, characterized by elevated Fe²⁺, MDA, GSH, and ROS levels (P < 0.001), alongside reduced Nrf2, xCT, and GPX4 expression. Mechanistically, DEHP upregulated miR155-5P, which directly targeted and inhibited ETS1 (P < 0.001); ETS1 overexpression reversed ferroptosis by activating the Nrf2/xCT/GPX4 pathway. Animal experiments confirmed DEHP-induced placental abnormalities (e.g., sinusoidal dilation) and ferroptosis, which were reversed by miR155-5P inhibitor. This study identifies the miR155-5P/ETS1 axis as a key mediator of DEHP-induced placental ferroptosis, highlighting ferroptosis as a critical target for mitigating DEHP reproductive toxicity. These findings provided potential biomarkers and therapeutic targets for preventing DEHP-related pregnancy complications, guiding future research on environmental pollutant-induced reproductive disorders.