3-BROMOPYRUVATE DEACTIVATES HEPATIC STELLATE CELLS AND INHIBITS LIVER FIBROSIS BY INHIBITING GLYCOLYSIS AND TGF-1β /SMAD PATHWAY

Abstract

Background: Liver fibrosis involves the activation of hepatic stellate cells (HSCs). Quiescent hepatic stellate cell (qHSC) transformation into activated hepatic stellate cells (aHSCs) requires energy and glycolytic intermediates. Targeting HSC metabolic reprogramming might reverse liver fibrosis; however, we lack effective treatment for liver fibrosis. Methods: Using 3‑bromopyruvate (3-BrPA), a hexokinase 2 (HK2) inhibitor, blocks the production of energy and aerobic glycolysis intermediates. Cell proliferation, migration, and apoptosis were evaluated. The capacity of glycolysis was conducted by lactate assay and glucose assay. Rat liver fibrosis was induced by carbon tetrachloride (CCl₄), the mechanism by which 3‑BrPA might inhibit liver fibrosis was evaluated by in vitro and in vivo assays. Results: 3-BrPA's anti-fibrotic effects were achieved by interfering with energy metabolism, as illustrated by cellular glucose consumption and lactate production in cellular supernatants and glycolytic enzyme expression, thereby inhibiting glycolysis and transforming qHSCs to aHSCs. However, pretreatment of rat hepatic stellate cells with TGF-β rescued the inhibitory effect of 3‑BrPA by increasing their glycolytic capacity and increasing extracellular matrix deposition. Additionally, 3‑BrPA's anti-fibrotic effect was validated in CCl₄-induced liver fibrosis. Examination of rat livers confirmed 3-BrPA's anti-fibrotic effects in vivo. Conclusions: Our data suggest that 3-BrPA exerts its anti-fibrotic activity in HSCs and a rat liver fibrosis model by targeting glycolysis and the TGF-β/SMAD pathway.