Background Metabolic-associated fatty liver disease (MAFLD) is characterized by excessive hepatic fat accumulation and can lead to severe complications, highlighting the need for effective treatments. This study investigated the active constituents and pharmacological mechanisms of Chaihu Lizhong Formula (CHLZF) in combating MAFLD through serum pharmacology, network pharmacology, and in vivo validation. Methods Blood-derived constituents from CHLZF-treated rats were analyzed using ultra-performance liquid chromatography coupled with Orbitrap mass spectrometry (UPLC-Orbitrap-MS). Potential targets and pathways were identified via network analysis and molecular docking. A rat model of MAFLD induced by a high-fat diet was used to assess the therapeutic effects of CHLZF and the associated mechanistic pathways employing serum biochemistry, histology, immunofluorescence, and Western blotting. Results Forty-two blood-derived constituents were linked to 268 potential MAFLD-related targets. SRC, AKT1, HSP90AA1, STAT3, and ESR1 were identified as key therapeutic targets. Enrichment analysis indicated that CHLZF may influence pathways such as PI3K/AKT, MAPK, HIF-1, FoxO, and insulin signaling. In vivo , CHLZF markedly improved dyslipidemia, liver function, and reduced hepatocyte steatosis in MAFLD rats. Mechanistically, these effects were associated with decreased phosphorylation of the PI3K/AKT/mTOR pathway and coordinated alterations in autophagy markers (increased Beclin 1 and LC3-II, decreased p62). Conclusion CHLZF demonstrates therapeutic potential in a preclinical MAFLD model, with effects linked to modulation of autophagy and the PI3K/AKT/mTOR signaling pathway. The identified bioactive components and potential targets warrant further preclinical investigation and mechanistic exploration.
Zhang et al. (Fri,) studied this question.