Integrating non‐targeted metabolomics and toxicology networks to study the mechanism of Esculentoside A‐induced hepatotoxicity in rats

Abstract Esculentoside A (EsA) is a kind of triterpenoid saponins from the root tuber of Phytolacca acinosa Roxb. It has extensive medicinal activity, such as antibacterial, anti‐inflammatory, immune regulation, and cell proliferation inhibition. However, some researches suggested that EsA can cause hepatotoxicity, whose mechanism is not precise. To ensure the safety and reliability in the clinical use of Phytolacca acinosa Roxb., it is necessary to establish a rapid and accurate method to evaluate the toxicity, analyze and verify the toxicity mechanism of EsA. Therefore, this research explored the mechanism of hepatotoxicity induced by EsA in rats and analyzed endogenous metabolites' changes in rat plasma by combining network toxicology with non‐targeted metabolomics. We obtained 58 critical targets of EsA induced hepatotoxicity in rats based on the strategy of network toxicology, including albumin, mitogen‐activated protein kinase 1, Caspase‐3, etc. Many important pathways were obtained by Kyoto Encyclopedia of Genes and Genomes enrichment analysis, such as HIF‐1 signaling pathway, TNF signaling pathway, IL‐17 signaling pathway, and other concerning pathways. Sixteen biomarkers, including 5‐hydroxykynurenamine, N ‐acetylserotonin, palmitic acid, etc., were screened from rat plasma using Ultra‐performance liquid chromatography coupled with quadrupole time of flight mass spectrometry (UPLC‐Q‐TOF/MS), mainly involve Glycerophospholipid metabolism, Tryptophan metabolism, and other metabolic pathways. Further analysis showed that EsA may induce liver injury by activating oxidative stress and energy metabolism disorders, triggering inflammation and apoptosis.