Gexia Zhuyu decoction alleviates carbon tetrachloride-induced hepatic fibrosis in mice: Mechanistic insights from integrated network analysis and experimental validation analysis

• Key Functions and Pathways of GXZYD Against Hepatic Fibrosis Obtained by Network Pharmacology. • GXZYD inhibits CCl 4 -induced early hepatic fibrosis in mice via anti-angiogenesis. • GXZYD suppresses PI3K/AKT signaling, reducing pathological angiogenesis. Gexia Zhuyu Decoction (GXZYD) is a classic Chinese medicine formula with significant efficacy in alleviating fibrosis. Remarkably, despite the established link between anti - angiogenesis, the PI3K/AKT signaling pathway, and the advancement of hepatic fibrosis (HF), it is still uncertain whether GXZYD achieves its anti - fibrotic effects by regulating these pathways. This study aimed to verify the potential mechanism by which GXZYD fights against HF through network analysis and experiments. The results of the network analysis analyses indicated that phosphatidylinositol 3-kinase/protein kinase B (PI3K/AKT) signaling is a pivotal regulatory pathway through which GXZYD alleviates HF and may exhibit anti-angiogenesis properties and reduce liver damage and collagen fiber formation. First, the efficacy of GXZYD in the treatment of early - stage heart failure was established. Subsequently, during the subsequent analysis, it was found that GXZYD effectively alleviated liver injury and reduced the accumulation of collagen, with an effect similar to that of colchicine. This research noted a decrease in the quantities of alanine aminotransferase (ALT) and aspartate aminotransferase (AST), along with a reduction in the levels of type IV collagen (Col-IV), laminin (LN), hyaluronic acid (HA), and procollagen type III (PC-III). Further analysis using pathological staining revealed improvements in hepatocellular necrosis and fibrous septa formation. Additionally, GXZYD enhanced the expression of CD31 and LYVE1 in relevant blood vessels and reduced the fenestration of liver sinusoidal endothelial cells (LSECs). The utilization of the agonist 740Y-P negated the protective impacts of GXZYD on anti - angiogenesis, the lessening of liver impairment, and the suppression of liver collagen accumulation. GXZYD mitigated early - stage liver fibrosis by impeding the PI3K/AKT signaling pathway, minimizing liver damage, regulating collagen deposition, and rejuvenating sinusoidal capillaries.