Identification and mechanistic exploration of key anti‐inflammatory molecules in American ginseng: Impacts on signal transducer and activator of transcription 3 STAT3 phosphorylation and macrophage polarization

Abstract American ginseng (AG) has been reported to have anti‐inflammatory effects in many diseases, but the key molecules and mechanisms are unclear. This study aims to evaluate the anti‐inflammatory mechanism of AG and identify the key molecules by in vivo and in vitro models. Zebrafish was employed to assess the anti‐inflammatory properties of AG and the compounds. Metabolomics was utilized to identify potential anti‐inflammatory molecules in AG, while molecular dynamics simulations were conducted to forecast the interaction capabilities of these compounds with inflammatory targets. Additionally, macrophage cell was employed to investigate the anti‐inflammatory mechanisms of the key molecules in AG by enzyme‐linked immunosorbent assay and western blotting. Seven potential anti‐inflammatory molecules were discovered in AG, with ginsenoside Rg1, ginsenoside Rs3 (G‐Rs3), and oleanolic acid exhibiting the strongest affinity for signal transducer and activator of transcription 3. These compounds demonstrated inhibitory effects on macrophage migration in zebrafish models and the ability to regulate ROS levels in both zebrafish and macrophages. The cell experiments found that ginsenoside Rg1, ginsenoside Rs3, and oleanolic acid could promote macrophage M2/M1 polarization ratio and inhibit phosphorylation overexpression of signal transducer and activator of transcription 3. This study revealed the key anti‐inflammatory molecules and mechanisms of AG, and provided new evidence of anti‐inflammatory for the scientific use of AG.