Combination of silybin and carvedilol synergistically alleviates liver fibrosis by inhibiting Wnt/β-catenin signaling

Liver fibrosis is a chronic and multifactorial liver disease that usually involves long-term hepatic injury, and is triggered mainly by hepatic stellate cells (HSCs) activation. Due to the complexity of liver fibrosis pathophysiology, the efficacy of monotherapies is limited. To overcome this limitation, the development of combination regimens has emerged as a potentially powerful strategy for treating liver fibrosis. In this study, a combination regimen involving silybin, a natural compound widely used for various liver diseases, with powerful antifibrotic efficacy was identified. The effects of 397 FDA-approved drugs in combination with silybin were evaluated using the COL1A1 luciferase reporter. Among the pairs of drugs, silybin and carvedilol had the strongest synergistic inhibitory effect on HSC activation. This synergistic effect was further validated in primary hepatic stellate cells (pHSCs) in vitro, and in CCl₄-treated mice in vivo. The ratio with the strongest synergistic and inhibitory effects was systemically evaluated to identify the optimal fixed-dose combination (50:1 for silybin and carvedilol in vivo). This combination regimen significantly inhibited CCl₄-induced liver fibrosis in a dose-dependent manner. Mechanistically, combined silybin and carvedilol inhibited HSCs activation through synergistically suppressing Wnt4/β-catenin signaling. This study provides a clinically feasible combination regimen with powerful synergistic and inhibitory effects on liver fibrosis.