Abstract
Assisted Reproductive Technologies (ART) are essential for addressing infertility and improving reproductive outcomes in both humans and animals. In vitro culture (IVC) supports embryo development outside the body but is often compromised by oxidative and endoplasmic reticulum (ER) stress, leading to impaired development and apoptosis. Demethylfuropinnarin (DMFP), a furocoumarin from Notopterygium incisum, has not been previously studied for its biological activity. Given the antioxidant properties of furocoumarins, this study aimed to determine whether DMFP could alleviate tunicamycin (TM)-induced oxidative and ER stress in porcine pre-implantation embryos. Our results show that DMFP significantly reduced reactive oxygen species (ROS) and preserved mitochondrial function. Additionally, DMFP increased Nuclear factor erythroid 2–related factor 2 (Nrf2) expression and decreased Kelch-like ECH-associated protein 1 (Keap1), leading to higher superoxide dismutase (SOD) and catalase (CAT) activity. DMFP treatment also improved cleavage and blastocyst formation rates, reduced apoptosis, and potentially alleviated ER stress, as indicated by lower levels of key ER stress markers. This study concludes that DMFP effectively reduces oxidative stress and may mitigate ER stress, enhancing embryo viability during IVC. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1038/s41598-026-38755-6.