Multi-parameter magnetic resonance imaging of zebularine in liver fibrosis treatment and calcineurin/NFAT3 mechanism.

BACKGROUND: Hepatic stellate cell (HSC) activation is key to liver fibrosis. Targeting DNA methylation shows promise. Zebularine, a methylation inhibitor, may suppress HSC activation via the calcineurin (CaN)/NFAT3 pathway. Magnetic resonance imaging (MRI) is a noninvasive tool for evaluating liver fibrosis evaluation tool, but multiparametric MRI for zebularine's effects in liver fibrosis mouse models has not been studied.

AIM: To clarify the anti-fibrosis mechanism and MRI-evaluated efficacy of zebularine.

METHODS: In vitro, transforming growth factor (TGF)-β1-stimulated human HSCs (LX-2) were treated with zebularine. α-smooth muscle actin, fibrotic and anti-fibrotic gene levels, and regulator of calcineurin1 (RCAN1) regulation were measured. In vivo, carbon tetrachloride (CCl4)-induced liver fibrosis in mice was treated with zebularine, and fibrosis was evaluated using various biochemical, histopathological, and MRI methods.

RESULTS: Zebularine upregulated RCAN1.4 protein (P < 0.01) and inhibited the CaN/NFAT3 pathway (P < 0.05). In HSCs, TGF-β1 reduced anti-fibrotic gene massage RNA (mRNA) and increased fibrotic mRNA (P < 0.05), whereas zebularine had the opposite effects (P < 0.01, P < 0.05). CCl4-treated mice exhibited increases in various fibrosis-related indices, all of which were reversed by zebularine treatment (P < 0.05).

CONCLUSION: Zebularine may reduce LX-2 activation and extracellular matrix deposition via RCAN1.4 and CaN/NFAT3 pathways. Multiparametric MRI can assess its efficacy, suggesting zebularine's potential as a liver fibrosis treatment.