Integrating Network Pharmacology and Experimental Validation to Elucidate the Role of Huaier in Attenuating Liver Fibrosis via the AKT Signalling Pathway

Liver fibrosis, a critical pathological precursor of cirrhosis and hepatocellular carcinoma, represents a significant unmet global health challenge because of the lack of effective targeted therapies that can reverse established fibrotic scarring. An integrated strategy was employed. Network pharmacology was used to identify potential targets, followed by molecular docking and dynamics simulations. In vivo validation utilized two established murine models: carbon tetrachloride (CCl4) and bile duct ligation (BDL)-induced fibrosis. In vitro studies employed human LX-2 hepatic stellate cells to assess the antifibrotic effects on myofibroblasts. Huaier administration significantly attenuated liver fibrosis, improved liver function and reduced collagen deposition in both animal models. Histopathological analysis confirmed diminished inflammatory infiltration and fibrotic scarring. In vitro, Huaier suppressed LX-2 cell proliferation and migration. Bioinformatics and simulation analyses suggested AKT1 as a central target, with high-affinity binding with the bioactive steroidal components of Huaier. Mechanistically, Huaier specifically inhibited the phosphorylation and activation of the AKT signalling pathway in hepatic myofibroblasts. This study provides the first compelling evidence that Huaier granules alleviate liver fibrosis by targeting the AKT pathway to inhibit myofibroblast activation. These findings illuminate its mechanistic basis and suggest that Huaier is a promising, multitarget therapeutic candidate for combating fibrotic liver disease.