Fuente: PubMed "essential oil" J Ethnopharmacol. 2026 May 8:121828. doi: 10.1016/j.jep.2026.121828. Online ahead of print.ABSTRACTETHNOPHARMACOLOGICAL RELEVANCE: Pimpinella diversifolia DC. is a traditional medicinal herb used in She and Miao ethnic medicine. Its root essential oil (PDREO) has demonstrated hepatoprotective effects, but its active constituents and the underlying mechanisms against acute liver injury (ALI) remain unclear.AIM OF THE STUDY: This study aimed to systematically identify the key anti-inflammatory and antioxidant components in PDREO and to elucidate their molecular mechanisms of action against ALI.MATERIALS AND METHODS: The protective effect of PDREO was evaluated in a D-galactosamine/ lipopolysaccharide (D-GalN/LPS)-induced ALI mouse model. PDREO was fractionated via silica gel chromatography, and the anti-inflammatory and antioxidant activities of the fractions and PDREO were assessed in LPS-stimulated RAW264.7 macrophages. Bioactive components were screened using gas chromatography-mass spectrometry (GC-MS) fingerprinting combined with grey relational analysis. Network pharmacology and molecular docking were employed to predict targets, followed by validation using cellular thermal shift assay (CETSA) and bio-layer interferometry (BLI). The mechanisms of the lead compound were further investigated in vitro and in vivo.RESULTS: PDREO significantly ameliorated D-GalN/LPS-induced liver damage, inflammation, and oxidative stress in mice. Six bioactive compounds were identified, among which 4-methoxy-2-(3-methyloxiranyl)phenyl isobutyrate (compound P59) exhibited the most potent activity. Network pharmacology and molecular docking predicted glycogen synthase kinase 3β (GSK-3β) as a key target. CETSA and BLI confirmed the direct binding of P59 to GSK-3β. Mechanistically, P59 promoted the inhibitory phosphorylation of GSK-3β, subsequently suppressed NF-κB p65 nuclear translocation, and downregulated pro-inflammatory cytokines. These effects were consistently observed in both LPS-stimulated macrophages and the D-GalN/LPS-induced ALI mouse model. Furthermore, p65 overexpression abrogated the inhibitory effect of P59 on pro-inflammatory cytokine secretion, validating NF-κB as the essential downstream effector of the GSK-3β/NF-κB axis.CONCLUSION: This study reveals that PDREO alleviates ALI through its anti-inflammatory and antioxidant properties. Compound P59 is identified as a pivotal active constituent that exerts hepatoprotection by directly targeting GSK-3β and inhibiting the GSK-3β/NF-κB signaling pathway. Our findings provide a pharmacological basis for the traditional use of P. diversifolia and highlight P59 as a promising candidate for ALI drug development.PMID:42107750 | DOI:10.1016/j.jep.2026.121828