Bensulide exposure causes cell division cycle arrest and apoptosis in porcine trophectoderm and uterine luminal epithelial cells

Abstract

As the use of herbicides in agriculture has increased worldwide, the importance of identifying unexpected toxic effects on non-target organisms is emerging. Bensulide is used on various agricultural crops as an organophosphate herbicide; however, it can pose a high risk to non-target organisms because of its long half-life and accumulative potential. Despite its high risk, the hazardous effects of bensulide on implantation and mechanisms in cells have not been reported. Therefore, in this study, intracellular mechanisms and potential risk of implantation failure were identified in porcine trophectoderm (pTr) and uterine luminal epithelial (pLE) cells derived from pigs with human-like molecular mechanisms in implantation. The LC50 values of bensulide were 5.21 mg/L in pTr cells and 6.49 mg/L in pLE cells. Both cell lines were exposed to bensulide at concentrations <5 mg/L in subsequent experiments. Treatment with 5 mg/L bensulide activated ERK1/2 and JNK. Disrupted mitochondrial membrane potentials of both cell types were identified. In addition, mitochondrial Ca2+ concentration increased to 261.24% and 228.04% in pTr and pLE cells, respectively, and cytoplasmic Ca2+ concentrations decreased by approximately 50% in both cell types. The abnormal regulation of various intracellular environments by bensulide causes cell division cycle arrest and apoptosis. Finally, 5 mg/L bensulide inhibited transcription of implantation-related genes. Collectively, our results suggest that bensulide may interrupt implantation during early pregnancy by disrupting maternal-fetal interaction.