Abstract:

This study investigated the mechanism of Chrysanthemi Indici Flos (CIF) against acetaminophen (APAP)-induced drug-induced liver injury (DILI) through integrated network pharmacology and in vivo experiments. Network pharmacology approaches were employed to predict bioactive components, therapeutic targets, and signaling pathways of CIF against DILI. Molecular docking validation (AutoDock Vina 1.1.2) confirmed binding affinity between key CIF components and core targets. Subsequent animal experiments elucidated the hepatoprotective effects and underlying mechanisms of CIF in APAP-induced liver injury models. Network pharmacology analysis identified luteolin, apigenin, and quercetin as the primary active constituents of CIF. The core therapeutic targets were found to be serine/threonine kinase 1 (AKT1) and cysteinyl aspartate-specific proteinase-3 (Caspase-3), which are closely associated with interleukin-17 (IL-17) and tumor necrosis factor (TNF) signaling pathways. Molecular docking confirmed stable binding interactions between these key CIF components and the core targets (Caspase-3 and AKT1). Animal experiments demonstrated that CIF administration significantly attenuated APAP-induced liver injury, as evidenced by reduced serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), lactate dehydrogenase (LDH), total bilirubin (TBIL), and direct bilirubin (DBIL), along with decreased hepatic malondialdehyde (MDA) levels. Conversely, CIF enhanced hepatic antioxidant capacity by elevating total superoxide dismutase (T-SOD) and glutathione (GSH) levels. Histopathological analysis revealed that CIF markedly ameliorated liver tissue damage, suppressed inflammatory cell infiltration, and inhibited hepatocyte apoptosis. RT-qPCR analysis revealed that CIF significantly downregulated the expression of Caspase-3, IL-6 and IL-1β genes in liver tissues of DILI mice. Western blot analysis demonstrated that CIF markedly upregulated B-cell lymphoma 2 (BCL-2) protein expression while downregulating Caspase-3 and BCL-2-associated X protein (BAX) levels in the liver tissues of DILI mice. These findings suggest that CIF mitigates APAP-induced liver injury by suppressing apoptosis, potentially through modulation of the BCL-2/BAX/Caspase-3 signaling pathway.

Key words: Chrysanthemi Indici Flos, acetaminophen, BCL-2/BAX/Caspase-3 signaling pathway