Stevioside Alleviates Con A-Induced Liver Injury by Activating the AMPK/NRF2 Signaling Pathway

Stevioside is a natural sweetener extracted from the perennial herb <i>Stevia rebaudiana</i> and has been approved by the FDA as an important dietary supplement. In addition, it possesses anti-inflammatory, antioxidant, and antiapoptotic properties. Nevertheless, its therapeutic efficacy against concanavalin A (Con A)-induced liver injury remains unclear. This study was designed to investigate the protective effect of stevioside on Con A-induced liver injury and to elucidate the underlying mechanisms. Using a Con A-induced mouse liver injury model, we evaluated liver damage via serum biochemistry and histopathology, identified key pathways through hepatic transcriptomics, and validated direct targets with molecular docking, CETSA, and DARTS assays. Compared with the model group, stevioside treatment dose-dependently reduced serum levels of hepatic injury markers and markedly ameliorated histopathological damage. Moreover, stevioside attenuated Con A-triggered hepatitis and hepatocyte apoptosis. Hepatic transcriptomic analyses revealed that differentially expressed genes were enriched in processes related to oxidoreductase activity, and stevioside effectively alleviated hepatic oxidative stress. Molecular docking, CETSA, and DARTS assays further identified AMPK may be the critical target for stevioside. In vitro experiments showed that stevioside suppressed Con A-induced inflammation and oxidative stress in RAW264.7 cells. In vivo, genetic knockout of Nrf2 or pharmacological inhibition of AMPK markedly diminished the hepatoprotective effects of stevioside. Collectively, stevioside activated the AMPK/NRF2 signaling pathway through its interaction with AMPK, thereby exerting hepatoprotective effects. Our study not only provides a theoretical basis for the application of stevioside in preventing liver diseases, but also opens new avenues for the treatment of liver injury.