Licorice Total Flavonoids Alleviate Liver Fibrosis by Inhibiting the TGF-β1 Signaling Pathways.

Liver fibrosis is a reversible pathological condition caused by chronic liver injury and represents a necessary stage leading to cirrhosis and hepatocellular carcinoma. Currently, liver fibrosis lacks specifically licensed therapeutic drugs. Licorice (Glycyrrhiza uralensis Fisch.) is a multi-efficacious traditional Chinese medicine, with licorice total flavonoids (LTF) serving as its primary active components, whereas the anti-hepatic fibrosis potential of LTF remains poorly elucidated. This study analyzed the core components of LTF using network pharmacology to investigate its anti-fibrotic targets and signaling pathways. Molecular docking and liquid chromatography-mass spectrometry (LC-MS) analysis, following BLI fishing, were used to systematically examine the active components and molecular mechanisms of LTF in the treatment of liver fibrosis. Network pharmacology analysis showed that LTF exerted anti-fibrotic effects by regulating multiple targets and pathways, with the TGF-β signaling pathway acting as a key hub. In human hepatocyte-derived stellate cells (LX-2), 16, 32, and 64 µg/mL LTF inhibited transforming growth factor-β1 (TGF-β1) induced activation of LX-2 cells and reduced the mRNA and protein expression of α-smooth muscle actin (α-SMA), Collagen I, and TGF-β1. LC-MS results from high-throughput bio-layer interferometry (BLI) showed that, compared with the control group, 95 flavonoid active components in the total licorice flavonoids group could serve as candidate compounds for binding TGF-β1, a core target of liver fibrosis. These results indicated that LTF may exert anti-fibrotic effects by regulating TGF-β-related signaling pathways via inhibiting TGF-β1 expression. Moreover, BLI analysis provides a reliable and feasible strategy for elucidating the mechanisms of traditional Chinese medicine in the treatment of hepatic fibrosis.