Pueraria Flos Alleviates Alcoholic Liver Injury Via Regulation of PPARα and MAOA: Deciphering the Effective Forms and Potential Mechanism Based on a Bioinformatics-Integrated Metabolic Profile Strategy and Experimental Validation

Abstract Background: Pueraria Flos, a representative medicinal and edible antidote for alcoholism, has rich clinical experience and remarkable curative effect in the treatment of alcoholic liver disease (ALD). However, its effective forms and hepatoprotective mechanism are remained unknown. Methods: A strategy based on UPLC-QTOF-MS combined with mass defect filtering technique was established for comprehensively identifying prototypes and metabolites absorbed and excreted into rat plasma, urine, bile and feces after oral administration. Then, the absorbed constituents with a relative high level were subjected to the network pharmacology, functional enrichment analysis and molecular docking to clarify the potential mechanism in the treatment of ALD. Furthermore, the therapeutic effect of PF on ALD and predicted mechanisms was further evaluated using a rat model of alcohol-induced liver injury and Western blot analysis. Results: 25 absorbed prototype constituents and 82 metabolites were identified or tentatively characterized with glucuronidation, sulfation, methylation, hydroxylation and reduction as their major metabolic pathways. The constructed absorbed constituent-target-pathway-disease network and docking analysis revealed that 4 metabolic components Te-7XG, genistein-7G-4'S, tectoridin-4'S and Te-7XG-4'S, 2 targets MAOA and PPARA, and 6 pathways related to lipid regulation and amino acid metabolism may play crucial roles in the PF mediated protection against ALD. An in vivo validation in rat further demonstrated that PF alleviated liver injury via activating and suppressing the PPARA and MAOA expression, respectively. Conclusions: The present results not only increase the understanding on the effective form and molecular mechanism of PF mediated protection against ALD, but also promote better application of PF as supplement food and herbal medicine for the treatment of ALD.