Lificiguat inhibits the collagen production of hepatic stellate cells independently on sGC activity

Background Liver fibrosis is driven by activated hepatic stellate cells (HSCs) that overproduce extracellular matrix, particularly collagen. Lificiguat, a soluble guanylate cyclase (sGC) stimulator, exhibits anti-fibrotic potential, but its mechanism in HSC activation remains unclear. This study aims to investigate the anti-fibrotic effect and mechanisms of lificiguat . Methods human HSCs are treated with different concentrations of lificiguat. Cell proliferation was assessed by CCK-8 assay and EdU incorporation assay. Fibrogenic markers of hepatic stellate cell including COL1A1, ACTA2 and TIMP1 are measured with RT-qPCR and Western blot. sGCβ1 (GUCY1B1) or ATG5 knockdown of HSCs are achieved with lentivirus transduction. Bulk RNA sequencing of HSC cells is performed to investigate the differentially expressed genes associated with lificiguat treatment. Serum ALT and AST, hepatic gene expression, and liver histology including Masson and Sirius red staining are analyzed with samples from CCl₄-induced fibrotic mice with or without lificiguat treatment. Results Lificiguat significantly inhibits cell proliferation and COL1A1 expression of HSCs without obvious cytotoxicity. GUCY1B1 knockdown in HSCs doesn’t reverse lificiguat’s effects, which indicates the anti-fibrotic effect of lificiguat doesn’t rely on sGC activity. Lificiguat enhances autophagic flux, but ATG5 knockdown fails to recover COL1A1 expression of HSCs treated with lificiguat. RNA-seq data indicates lificiguat modulates JAK-STAT and IL-17 pathways of HSCs. Lificiguat reduced liver injury markers including serum ALT and AST in CCL₄-challenged mice. In addition, lificiguat reduces mRNA expression of fibrogenic marker gene including Col1a1 and Acta2 and attenuate liver fibrosis in CCl₄ mice models. Conclusion Lificiguat attenuates liver fibrosis by inhibiting HSC proliferation and collagen synthesis through sGC- and ATG5-independent mechanisms, potentially via regulating JAK-STAT and IL-17 pathways.