Exploring the inhibitory effects of Guben Antai Decoction on oxidative stress damage and ferroptosis via the Nrf2/HO-1 pathway: A mechanistic study on improving Recurrent Spontaneous Abortion.

ETHNOPHARMACOLOGICAL RELEVANCE: Guben Antai Decoction (GBAT) is a traditional Chinese medicine formula that significantly increases embryo implantation rates and reduces early pregnancy loss in patients with recurrent spontaneous abortion (RSA).

AIM OF THE STUDY: This study aims to explore the influence and possible mechanism of GBAT in improving recurrent spontaneous abortion.

MATERIALS AND

METHODS

In vivo, normal pregnancy and RSA models were established using CBA/J female × BALB/c male and CBA/J female × DBA/2 male mouse pairings, respectively. GBAT (18.05 g/kg/d) or dydrogesterone (DYD; 3.03 mg/kg/d) was intragastrically administered to pregnant mice in the RSA model from gestational day 1. An oxidative stress model was established in vitro by treating human trophoblast HTR-8/SVneo cells with H2O2. Placental villi damage was assessed using hematoxylin and eosin staining, and mitochondrial morphology was examined using transmission electron microscopy. The expression of GPX4, ACSL4, SLC7A11, Nrf2, and HO-1 were measured using RT-qPCR, western blotting, and immunohistochemistry. Lipid peroxidation markers ROS, GSH, GPx, MDA, and Fe2+ were quantified using commercial kits.

RESULTS

In mice with RSA, ferroptosis occurred in the placental villous tissue. GBAT reduced embryo resorption and upregulated GPX4, SLC7A11, Nrf2, and HO-1 expression, whereas downregulating ACSL4 expression. In vitro, H2O2-induced ferroptosis decreased trophoblast viability and caused ROS accumulation and lipid peroxidation. GBAT attenuated these effects by activating Nrf2. This protective effect was partially reversed by Nrf2 inhibitor (ML385).

CONCLUSION

This study is the first to demonstrate that GBAT mitigates ferroptosis in trophoblasts by activating the Nrf2/HO-1 pathway, providing new mechanistic insights into its potential for treating RSA.