UPLC-FT-ICR-MS and mechanistic study reveals the protective effect of Callistephus chinensis flower against alcoholic liver disease

• UPLC-FT-ICR-MS analysis identified 23 major compounds in CCF, primarily flavonoids and phenolic acids. • CCF can effectively reduce inflammatory reaction and inhibit lipid accumulation in liver. • CCF mainly prevents alcoholic liver injury by regulating AMPK and TLR4/NF-κB signaling pathway. • CCF may be a candidate drug for prevention and treatment of ALD. Liver disease caused by excessive alcohol consumption is a serious threat to human health. Previous studies have shown that Callistephus chinensis flower (CCF) can ameliorate alcoholic liver disease (ALD), but its mechanism remains unreported. The chemical composition of CCF was identified in vitro by employing ultra-performance liquid chromatography coupled with Fourier transform ion cyclotron resonance mass spectrometry (UPLC-FT-ICR-MS). C57BL/6J mice were divided into control, ethanol, CCF extract (100, 200, 400 mg/kg), and silymarin (100 mg/kg) groups. Hepatic pathology and lipid infiltration were assessed via H&E and Oil Red O staining. Commercial kits were used to measure TC, TG, AST, ALT, MDA, and GSH-Px levels. IHC was performed for PPARα, IL-1β, F4/80, and TLR4, while Western blot quantified AMPKα, p-AMPKα, PPARα, CPT1α, ACC, and p-ACC. LC-MS/MS preliminarily identified 23 major compounds, with flavonoids and phenolic acids being the predominant components in CCF extract. CCF administration upregulated p-AMPKα, PPARα, and CPT1α, and downregulated TLR4, p-NF-κB, and p-IκBα. It effectively reduced serum inflammatory factors, decreased lipid accumulation, promoted lipid metabolism, and alleviated liver injury by modulating AMPK and TLR4/NF-κB pathways. This study is the first to demonstrate that CCF ameliorates ALD via AMPK and TLR4/NF-κB pathways, providing a potential candidate for ALD treatment.