Abstract: Objective To investigate the therapeutic effects of Haizao and Gancao against oxidative stress and ferroptosis in goiter rats and the molecular mechanism. Methods Based on the network pharmacology platform, the potential mechanisms of action of Haizao and Gancao in ameliorating goiter were explored. Seventy-eight Wistar rats were randomly divided into a control group, model group, Euthyrox group, and low-, medium-, and high-dose groups of Haizao and Gancao (1.98、3.96、7.92 g·mL^-1). Except the control group, all the groups were induced to establish a goiter rat model using propylthiouracil (PTU) and underwent corresponding interventions. After 28 days, the condition of the rats and thyroid coefficient were observed, and indicators of oxidative stress and ferroptosis were detected. Results KEGG pathway and GO analyses were closely correlated with the regulation of redox homeostasis and lipid metabolism, with PTGS2 identified as a key target. Animal experiments suggested that rats in the model group manifested hypometabolic symptoms and structural disorganization of thyroid follicles. Haizao and Gancao of varying doses significantly ameliorated the health status and histopathology of goitrous rats, with the efficacy most pronounced in the low-dose group. An oxidative stress imbalance was observed in goitrous model rats. The oxidative stress index (OSI) (P<0.05) and reactive oxygen species (ROS) levels (P<0.001) were upregulated in the low-dose Haizao and Gancao group. Prussian blue staining and transmission electron microscopy pointed to characteristic iron deposition and ultrastructural changes of mitochondria in thyroid tissues in each group. The Nrf2/HO-1 signaling pathway was activated while the expressions of key markers of ferroptosis-GPX4 and PTGS2- (P<0.05, P<0.001) were regulated in the low-dose Haizao and Gancao group (P<0.01, P<0.05). Conclusion Haizao and Gancao can ameliorate goiter by regulating the ‘oxidative stress-ferroptosis’ signaling axis through the Nrf2/HO-1/GPX4 pathway.

Key words: Haizao, Gancao, Oxidative Stress, Ferroptosis, Nrf2/HO-1/GPX4, UPLC-MS/MS, Rat