Amelioration of Liver Fibrosis via In Situ Hepatic Stellate Cell Conversion Through Co-Inhibition of TGF-β and GSK-3 Signalling

Liver Int. 2025 Jul;45(7):e70187. doi: 10.1111/liv.70187.

Abstract

Background and aims: Liver fibrosis, a progressive condition driven by chronic liver injury and excessive scar tissue formation, can lead to cirrhosis, a life-threatening disease. Activation of hepatic stellate cells (HSCs) is central to fibrosis progression, yet current therapies fail to halt or reverse this process. This study evaluated a combination therapy targeting HSCs to ameliorate fibrosis and promote liver repair.

Methods: A small molecule cocktail, SBCH (SB431542, a TGF-β inhibitor, and CHIR99021, a GSK-3 inhibitor), was tested in three fibrosis models: CCl4-induced, bile duct ligation (BDL) and non-alcoholic steatohepatitis (NASH) with diethylnitrosamine (DEN). Therapeutic effects were assessed using phenotypic analyses, in vivo tracing and single-cell RNA sequencing to uncover mechanisms.

Results: SBCH significantly reduced fibrosis in all models by inhibiting HSC activation and fibrogenic activity. The suppression of PI3K/Akt pathway and EMT cascade contribute to the fibrosis-ameliorating effect of SBCH treatment. Furthermore, in vivo tracing and single-cell RNA sequencing revealed that SBCH induced the conversion of activated HSCs into hepatocyte-like cells (ciHeps), which integrated into liver tissue, repaired liver damage and restored liver integrity and function.

Conclusions: SBCH mitigates liver fibrosis through multifaceted mechanisms, including the inhibition of HSC activation, suppression of fibrogenic activity and regulation of key signalling pathways such as PI3K/Akt and EMT. In addition, SBCH induces the trans-differentiation of activated HSCs into hepatocyte-like cells (ciHeps), effectively reducing pathogenic HSCs while increasing functional ciHeps. This dual-target approach not only facilitates liver tissue repair but also restores liver function, offering a promising therapeutic strategy for liver fibrosis and cirrhosis, with potential applications in conditions arising from various aetiologies of liver injury.

Keywords: hepatic stellate cells; hepatocytes; lineage tracing; liver fibrosis and cirrhosis; trans‐differentiation.

MeSH terms

  • Animals
  • Benzamides* / pharmacology
  • Benzamides* / therapeutic use
  • Carbon Tetrachloride
  • Cell Transdifferentiation / drug effects
  • Diethylnitrosamine
  • Dioxoles* / pharmacology
  • Dioxoles* / therapeutic use
  • Disease Models, Animal
  • Glycogen Synthase Kinase 3* / antagonists & inhibitors
  • Glycogen Synthase Kinase 3* / metabolism
  • Hepatic Stellate Cells* / drug effects
  • Hepatic Stellate Cells* / metabolism
  • Liver / drug effects
  • Liver / pathology
  • Liver Cirrhosis* / drug therapy
  • Liver Cirrhosis* / pathology
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Non-alcoholic Fatty Liver Disease / complications
  • Non-alcoholic Fatty Liver Disease / drug therapy
  • Pyridines* / pharmacology
  • Pyridines* / therapeutic use
  • Pyrimidines* / pharmacology
  • Pyrimidines* / therapeutic use
  • Signal Transduction / drug effects
  • Transforming Growth Factor beta* / antagonists & inhibitors
  • Transforming Growth Factor beta* / metabolism

Substances

  • Transforming Growth Factor beta
  • Benzamides
  • Chir 99021
  • 4-(5-benzo(1,3)dioxol-5-yl-4-pyridin-2-yl-1H-imidazol-2-yl)benzamide
  • Dioxoles
  • Pyridines
  • Pyrimidines
  • Glycogen Synthase Kinase 3
  • Carbon Tetrachloride
  • Diethylnitrosamine