Gandouling inhibits hepatic fibrosis in Wilson's disease through Wnt-1/β-catenin signaling pathway

Wilson's disease (WD) hepatic fibrosis is the result of chronic liver injury induced by Cu2+ deposition in the liver. Gandouling (GDL) is a hospital preparation of the First Affiliated Hospital of Anhui University of Chinese Medicine. Previous studies have found that GDL can play an anti-inflammatory, anti-oxidation, and promote Cu2+ excretion, which has a clear anti-WD effect. We found that Wnt-1 was significantly up-regulated in the liver tissue of toxic-milk (TX) mouse in the WD gene mutant model, and the monomer components of GDL could combine well with Wnt-1. Therefore, in this work, we used RT-qPCR, Western blot, immunofluorescence, network pharmacology, molecular docking, and related methods to study the effects of GDL on hepatic stellate cell (HSC) activation and Wnt-1/β-catenin pathway in TX mice to clarify the effect of GDL on WD hepatic fibrosis. GDL could alleviate hepatic fibrosis, improve liver function, and inhibit the activation of HSC in TX mice. Network pharmacology predicted that the Wnt-1/β-catenin was the target of GDL, and molecular dynamics further revealed that GDL has a good binding ability with Wnt-1 and inhibits the Wnt/β-catenin signaling pathway through Wnt-1. Furthermore, we found that GDL blocked the Wnt-1/β-catenin signaling pathway in the liver of TX mice in vivo. In vitro, serum containing GDL blocked the Cu2+ ion-induced Wnt-1/β-catenin signaling pathway in LX-2 cells. Therefore, GDL blocked the Wnt-1/β-catenin signaling pathway, inhibited HSC activation, and improved WD hepatic fibrosis by binding to Wnt-1. GDL improves hepatic fibrosis in WD model mice by blocking the Wnt-1/β-catenin signaling pathway, and Wnt-1 may be a new target for the diagnosis and treatment of WD. This reveals a new mechanism of GDL against WD, and promotes the clinical promotion of GDL. Wnt-1 expression was significantly up-regulated in TX mice and Cu2+-induced LX-2 cells. Wnt-1 significantly activated the Wnt/β-catenin signaling pathway, promoted the activation of HSC, increased the levels of inflammatory cytokines TNF-α, IL-1β, and IL-6, and significantly increased the expression of α-SMA and collagen 1. GDL has an obvious therapeutic effect on TX mice. Combined with the molecular docking results, we found that GDL has a good binding ability with Wnt-1 and inhibited the Wnt/β-catenin signaling pathway through Wnt-1. This indicates that GDL exerts antifibrotic effects through the Wnt-1/β-catenin signaling pathway. [Display omitted] •Wnt-1 in Wil