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

Liver fibrosis, a critical pathological precursor to cirrhosis and hepatocellular carcinoma, represents a significant unmet global health challenge due to the lack of effective targeted therapies that can reverse established fibrotic scarring. An integrated strategy was employed. Network pharmacology identified 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 anti-fibrotic 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 pinpointed AKT1 as a central target, showing high-affinity binding with Huaier's bioactive steroidal components. Mechanistically, Huaier specifically inhibited the phosphorylation and activation of the AKT signaling pathway in hepatic myofibroblasts. This study provides the first compelling evidence that Huaier granule alleviates liver fibrosis by targeting the AKT pathway to inhibit myofibroblast activation. These findings illuminate its mechanistic basis and reposition Huaier as a promising, multi-targeted therapeutic candidate for combating fibrotic liver disease.