Integrating network pharmacology and in vivo evaluation reveals chicory (Cichorium intybus L.) extract activates autophagy to alleviate fatty liver hemorrhagic syndrome in laying hens.

Fatty liver hemorrhagic syndrome (FLHS) is a nutritional metabolic disease in poultry characterized by disrupted lipid metabolism in the liver. This study aimed to investigate the pharmacological effects and mechanisms of chicory (Cichorium intybus L.) root ethanol extract (CEE) on HELP-diet-induced FLHS. Seventy-two 50-week-old Hy-Line Brown laying hens were randomly allocated into three groups, with four replicates in each group and six hens per replicate. A standard basal diet was provided to the negative control (NC) group, whereas the other two groups were fed the high-energy low-protein (HELP) diet supplemented with 0 g/kg or 15 g/kg CEE, designated as the FLHS and CEE groups, respectively. The adaptation period lasted for 1 week, followed by a 12-week experimental period. UPLC-MS/MS-based component profiling of CEE, combined with network pharmacology analysis, identified three bioactive compounds: lactucopicrin, 2S,2'S-Aurantiamide acetate, and luteolin. Body weight was significantly increased by the HELP diet, while hens receiving CEE markedly lower body weight relative to the FLHS group at week 12 (P < 0.05 or 0.01). CEE supplementation significantly restored the laying rate, and reduced the Feed/egg ratio (P < 0.01). The FLHS group showed pronounced hepatic steatosis in laying hens, with increased TG and TC levels in serum and liver and elevated ALT and AST activities, which were markedly reversed by CEE treatment. There were 93 metabolites showing significant differences in the liver between the FLHS and CEE groups, such as 1-stearoyl-2-arachidoylglycerol, PE-NMe2, and 19-hydroxyarachidonic acid. Differentially expressed metabolites are enriched in the autophagy signaling pathway according to KEGG. CEE treatment significantly inhibited the protein levels of p-mTOR/mTOR, p62, Fasn, Acc, and Pparγ, while significantly increasing those of p-Ulk1/Ulk1, Atg5, LC3II/LC3I, and Pparα (P < 0.05 or 0.01). In conclusion, supplementation with 15 g/kg CEE effectively mitigated FLHS in laying hens by modulating hepatic lipid metabolism by activating autophagy. CEE represents a novel pharmacological strategy for FLHS treatment, highlighting chicory potential in poultry.