Kurarinol A from Sophora flavescens as potential anti-liver fibrosis agent that inhibits the activation of LX-2 cells by regulating TGF-β/Smads signaling pathway

Objective This work aims to discover bioactivity against liver fibrosis for the lavandulyl flavonoids from one botanical source of S. flavescens. Further use of transcriptomics technology to explore the molecular mechanism of anti-hepatic fibrosis. Methods In this study, a model of LX-2 cells activation induced by transforming growth factor-β1 (TGF-β1) was established. A total of 35 free phenolics were isolated from S. flavescens to form a small compound library. These compounds on the proliferation of LX-2 cells were screened using MTS method. Furthermore, cell scratch, transcriptomics technology, real-time quantitative polymerase chain reactions (RT-qPCR) and Western blotting were used to evaluate the inhibitory effect of lavandulyl flavonoids on the proliferation and activation of LX-2 cells, and to explore the mechanism of lavandulyl flavonoids in improving liver fibrosis. Results The results showed that a total of 11 compounds had a significant inhibitory effect on the proliferation of LX-2 cells and their IC ₅₀ was between 4-40 μM by MTS assay. Among them, 8 compounds were reported for the first time. Particularly, kurarinol A ( 1 , IC ₅₀ 12.65 μM) showed noticeable inhibitory activities. Furthermore, The results of cell scratch test showed that KA inhibited the migration of LX-2 cells. The migration process was carried out in a dose-dependent manner at 24 and 48 hours. Then, KA remarkably inhibit the mRNA and protein levels of liver fibrosis markers (α-SMA, fibronectin and collagen I), and could effectively inhibit the development of liver fibrosis. Additionally, transcriptome analysis revealed that there were 106 differentially expressed genes (DEGs) with remarkably expression differences by the treatment of KA were identified in the LX-2 cells. The mechanism studies elucidated that KA exerted protective activities involved in modulating the TGF-β/Smads signaling pathway. Among them, KA inhibited the gene and protein of TGF-β1, Smad2, Smad3, and Smad4 levels, respectively. Conclusion KA can improve liver fibrosis. The mechanism of its anti-hepatic fibrosis was achieved by regulating the TGF-β/Smads signaling pathway. KA could be an effective anti-fibrosis agent.