Investigating the Antifibrotic Action of <scp><i>Foeniculum vulgare</i></scp> Root Bark Volatile Oil Through the HK2/PKM2/LDHA Pathway

Foeniculum vulgare root bark (FVRB), a traditional ethnomedicine, demonstrates hepatoprotective properties and has been clinically employed in managing liver fibrosis. As volatile oils represent principal bioactive constituents of FVRB, the therapeutic potential of F. vulgare root bark volatile oil (FVRBO) against liver fibrosis remains unelucidated. This study investigates the antifibrotic mechanisms of FVRBO through integrated in vitro and in vivo approaches. The FVRBO was extracted using the steam distillation method, and the content of its main components was determined by gas chromatography. Liver fibrosis models were established using CCl₄-induced mice and TGF-β1-induced JS-1 cells. Antifibrotic efficacy was validated through significant downregulation of fibrotic markers and inflammatory cytokines. Integrated transcriptomic and metabolomic analyses revealed FVRBO's mechanism. The principal constituents of FVRBO were quantified as dillapiole, apiole, and myristicin, with the extraction method demonstrating consistent stability. In vivo, FVRBO significantly attenuated CCl₄-induced liver injury in mice, as evidenced by decreased liver and spleen indices, reduced serum ALT and AST levels, attenuated inflammatory infiltration, and suppressed collagen deposition in hepatic tissue. In vitro, FVRBO effectively inhibited TGF-β-induced JS-1 cell activation and downregulated fibrotic markers at mRNA and protein levels. Integrated transcriptomic-metabolomic analysis revealed FVRBO exerts antifibrotic efficacy by regulating the glycolysis pathway and suppressing the expression of HK2, PKM2, and LDHA. FVRBO exerts antifibrotic effects by modulating the glycolytic pathway and its associated gene expression. This study provides mechanistic evidence for FVRBO as a phytopharmaceutical candidate and establishes a foundation for developing natural-product-based anti-fibrotic therapeutics.