Network toxicology and cell experiments reveal the mechanism of saikochromone A-induced hepatotoxicity.

The potential toxicity of traditional Chinese medicine (TCM) remains a significant barrier to its broader clinical application. Bupleurum, a commonly used TCM, contains saponins and volatile oils that have been shown to cause hepatotoxicity. Due to the complex chemical composition of Bupleurum, saikochromone A (SA), another major bioactive component, has not been thoroughly assessed in terms of its toxicological profile and underlying mechanisms. Our study aimed to evaluate the potential hepatotoxic effects of SA by integrating network toxicology, molecular docking analysis, and in vitro experimental approaches. A total of 109 possible targets associated with SA-induced hepatotoxicity were identified through the publicly available online databases. Functional enrichment analysis revealed significant involvement of pathways related to chemical carcinogenesis (DNA adducts) and cytochrome P450-mediated metabolism. Molecular docking simulations further demonstrated strong binding affinities between SA and key proteins, including CYP2E1, CYP1A1, CYP3A4, PTGS2, and MMP-9. In vitro experiments showed that SA significantly reduced cell viability, with an IC50 value of 176.8 μM. Additionally, SA exposure was found to upregulate the expression levels of CYP2E1, CYP1A1, CYP3A4, PTGS2, and MMP-9. By combining computational and experimental methodologies, this study systematically elucidates, for the first time, that high concentrations of SA may induce hepatotoxicity primarily through the CYP450 enzyme system, inflammatory pathway, and matrix metalloproteinase activation. These findings offer a novel methodological framework for identifying and evaluating toxic components in TCM and contribute valuable insights into the potential adverse effects of TCM in clinical practice.