Ginsenoside Rk1 suppresses the epithelial-mesenchymal transition of hepatic stellate cells via PFKFB2-mediated aerobic glycolysis.

Liver fibrosis may progress to cirrhosis or hepatoma without treatment. Hepatic stellate cell (HSC) activation, which could be promoted by epithelial-mesenchymal transition (EMT) process, is crucial for liver fibrosis progression. Ginsenoside Rk1 (GRk1) is a ginsenoside with properties against inflammatory and tumor. Nonetheless, its effects on fibrosis remain unclear. In this study, the suppressive effects of GRk1 on HSC activation as well as liver fibrosis and its underlying mechanism were explored in CCl₄-treated liver fibrosis mice and primary HSCs. Molecular docking analysis verified the interaction between PFKFB2 and GRk1. In addition, RNA-sequence analysis and proteomic lactylation analysis were performed in GRk1-treated HSCs. The results showed that GRk1 attenuated CCl₄-induced liver fibrosis and HSC activation, with suppressed HSC EMT. Notably, it was revealed that GRk1 inhibited aerobic glycolysis and its production lactate via targeting PFKFB2, which was reversed by PFKFB2 overexpression. In addition, it was found that STAT3 lactylation participated in GRk1-inhibited HSC EMT and activation. Further experiments demonstrated that K161 site of STAT3 was responsible for GRk1-inhibited STAT3 lactylation. Moreover, GRk1 treatment led to reduced STAT3 nuclear expression, decreasing TGF-β1 expression and suppressing EMT process, ultimately inhibiting HSC activation. To sum up, this study reveals that GRk1 inhibits aerobic glycolysis via directly targeting PFKFB2, leading to the inhibition of STAT3 lactylation and nuclear translocation, which finally contributes to HSC EMT inhibition and inactivation.