Protective effects of atractylenolide III on diquat-induced hepatic injury via PI3K/AKT/mTOR pathway inhibition and autophagy enhancement in mice.

This study aimed to assess the protective effects of atractylenolide III (ATL-III) against diquat (DQ)-induced liver injury and its mechanisms. Bioinformatics analysis identified ROS accumulation and PI3K/AKT pathway activation as key features of DQ-induced toxicity. Network pharmacology predicted PI3K, AKT, and mTOR as targets of ATL-III. In C57BL/6J mice, serum ALT, AST, and TB were measured, and liver histopathology was evaluated by H&E staining. Protein expression of LC3, p62, and PI3K/AKT/mTOR pathway components was analyzed via western blot, with p62 localization examined by immunofluorescence. In HepG2 cells, cell viability, apoptosis (caspase-3/9 activity), ROS levels, and ALT/AST release were measured. Expression of LC3, p62, and PI3K/AKT/mTOR pathway proteins was also analyzed. Bioinformatics analyses indicated that DQ-induced hepatic injury is associated with elevated ROS production and activation of the PI3K/AKT pathway. In vivo, ATL-III administration reduced serum ALT, AST, and TB levels, alleviated hepatic inflammation, and improved histopathological features. ATL-III treatment downregulated p62, phosphorylated PI3K, AKT, and mTOR, and increased LC3II/I ratio, indicative of enhanced autophagic activity. In HepG2 cells, ATL-III improved cell survival, reduced apoptosis, decreased ROS accumulation, and lowered ALT/AST release. These in vitro findings corroborated the in vivo findings, further demonstrating downregulation of p62 and phosphorylation of PI3K/AKT/mTOR components, alongside increased LC3II/I conversion. ATL-III exerted protective effects against DQ-induced hepatic injury in both murine models and human hepatocyte-derived cells. These effects were associated with inhibition of the PI3K/AKT/mTOR signaling pathway, activation of autophagy, and reduction in intracellular ROS accumulation.