Cigarette smoke-induced trophoblast cell ferroptosis in rat placenta and the effects of L-arginine intervention

Cigarette smoke (CS) disrupts placental development, and impairs fetal health and maternal fertility, thus resulting in low birth weight, premature delivery, and spontaneous abortion; however, the underlying mecha-nisms remain unclear. This study investigated the mechanism through which CS impairs placental trophoblast cell viability and function. An in vivo study in pregnant rats exposed to CS indicated that CS-exposure decreased antioxidant factors expression and blocked NRF2 activation in the placenta. Anti-ferroptosis regulators expres-sion was downregulated, and pro-ferroptosis regulators expression was upregulated in placentas from CS -exposed rats. Further analysis revealed that cigarette smoke extract (CSE) led to peroxins downregulation and decreased the number of peroxisomes. An in vitro study in HTR-8/SVneo(HTR-8) cells showed that CSE led to free iron and ROS accumulation, and subsequently induced lipid peroxidation and cell death. Ferroptosis in-hibitors and the antioxidant L-arginine (ARG) partially inhibited CSE-induced cell death. ARG partially alleviated the toxic effects of CSE by promoting antioxidant factors expression in placenta and suppressing HTR-8 cell ferroptosis. Knockdown of PEX14, a peroxisome biogenesis marker, led to the downregulation of multiple PEXs, thus increasing intracellular H2O2 levels and inducing HTR-8 cell ferroptosis. These findings demonstrated that ferroptosis is responsible for CSE-induced trophoblast cell death and suggest that peroxisome dysfunction is involved in CSE-induced ferroptosis. Therefore, CSE-induced ferroptosis may serve as a potential therapeutic target for preventing adverse pregnancy outcomes.