Calycosin protects from histopathological and ultrastructural changes in ethanol-induced liver injury and exhibits predicted binding affinity to TNF-α/NF-κB/HIF-1α proteins.

BACKGROUND/OBJECTIVES: Liver fibrosis is the consequence of the wound-healing response of the liver to frequent injury. Calycosin is a flavonoid, which has been documented for its hepatoprotective properties. The aim of this study was to evaluate the hepatoprotective action of calycosin in a model of ethanol-induced liver injury (EILI) in rats. For mechanistic insights, we aimed to measure tumor necrosis factor-α (TNF-α), nuclear factor-κB (NF-κB), and hypoxia-inducible factor-1 alpha (HIF-1α) protein levels in liver samples and evaluate the ability of calycosin to bind these pathologic molecules. METHODS: Network pharmacology and molecular docking were performed for discovering the interacting target proteins. Forty male rats were assigned into five groups: 1) the vehicle, 2) the calycosin per se group, 3) the EILI control group, and 4) and 5) the EILI + calycosin (5 or 10 mg/kg) groups. The liver homogenates were used for ELISA measurement of TNF-α, NF-κB, and HIF-1α. Furthermore, liver specimens were used for histopathological and ultrastructural investigations. RESULTS: The network pharmacology study confirmed the role of TNF-α/NF-κB/HIF-1α signaling in EILI, and molecular docking explored the possible interaction between calycosin and these three molecules (docking score = -7.6, -7, and -7.6 kcal/mol). The rat study showed that calycosin was able to attenuate histopathological and ultrastructural changes in rat livers, reduce collagen accumulation, and prevent the increases shown in liver enzyme activities (2.17-fold for ALT and 2.26-fold for AST). CONCLUSION: Thus, these integrated in silico and in vivo studies confirmed the hepatoprotective effect of calycosin against the EILI rat model and provided a mechanistic insight through TNF-α/NF-κB/HIF-1α signaling. Further studies are warranted to fully explore the protective mechanism of calycosin in EILI.