Abstract: Objective To investigate the regulatory effect of Yangxue Antai prescription on placental ferroptosis in recurrent spontaneous abortion (RSA) mice. Methods Cell experiment was divided into three groups. In Control group, 10% blank serum was added. In Erastin group, 25 μmol·L^-1 Erastin was added to induce ferroptosis. In Erastin+Yangxue Antai group, 25 μmol·L^-1 Erastin was added and intervened with 10% Yangxue Antai serum. The protein expressions of ferritin heavy chain 1 (FTH1) and ferroportin 1 (FPN1) in JEG-3 cells were measured by Western blot. Animal experiment was divided into four groups with four animals in each group. Normal pregnancy group and RSA group were intragastrically administered with distilled water. Yangxue Antai group and RSA+Yangxue Antai group were intragastrically administered with 28.08 g·kg^-1 of Yangxue Antai prescription. The activity of superoxide dismutase (SOD), the contents of reduced glutathione (GSH), malondialdehyde (MDA) and ferrous ion (Fe²⁺) in mouse placenta were measured. The protein expression of glutathione peroxidase 4 (GPX4), solute carrier family 7, member 11 (SLC7A11), heme oxygenase 1 (HO-1), FTH1, FPN1, transferrin receptor protein 1 (TfR1) and divalent metal transporter 1 (DMT1) in mouse placenta were measured by Western blot. Results Compared with the control group, the protein expressions of FTH1 and FPN1 in JEG-3 cells in the Erastin group were significantly decreased. Compared with the Erastin group, the protein expressions of FTH1 and FPN1 in JEG-3 cells in the Erastin+Yangxue Antai group were significantly increased. Compared with the normal pregnancy group, the SOD activity, GSH and Fe²⁺ contents in placentas in the RSA group were significantly decreased, but the MDA content was significantly increased. The protein expressions of GPX4, SLC7A11, HO-1, FTH1 and FPN1 were significantly decreased in placentas, while those of TfR1 and DMT1 were significantly increased. Compared with the RSA group, the SOD activity, GSH and Fe²⁺ contents in placentas in the RSA+Yangxue Antai group were significantly increased, while that of MDA was significantly decreased. The protein expressions of GPX4, SLC7A11, HO-1, FTH1 and FPN1 were significantly increased in placentas while those of TfR1 and DMT1 were significantly decreased. Conclusion Yangxue Antai prescription can alleviate oxidative stress, reduce the intake of Fe²⁺, increase the storage and transfer of Fe²⁺, thus inhibiting the accumulation of Fe²⁺ and inhibiting the occurrence of ferroptosis.

Key words: Yangxue Antai Prescription, TCM Formula, Recurrent Spontaneous Abortion, Ferroptosis, Oxidative Stress, Mice, Rats, Placenta