Integrative Network Pharmacology and Experimental Validation Reveal Multi-target Hepatoprotective Mechanisms of Bioactive Compounds from Xiao-Yao-San

AuthorZi-Yi Zhengen
AuthorJing-Yi Chenen
AuthorRoy You Chen Quahen
AuthorGuo-Hua Dingen
AuthorJing Chenen
OrcidZi-Yi Zheng [0009-0008-0220-8653]en
OrcidJing-Yi Chen [0000-0002-1218-6389]en
OrcidRoy You Chen Quah [0000-0002-3014-8976]en
OrcidGuo-Hua Ding [0000-0002-2484-5855]en
OrcidJing Chen [0000-0001-8697-3265]en
Issued Date2026-12-31en
AbstractBackground: Xiao-Yao-San (XYS), a classical Traditional Chinese Medicine formula documented in the Chinese Pharmacopoeia, has long been used to treat hepatic disorders; however, its multi-target hepatoprotective mechanisms remain systematically uncharacterized. Objectives: This study aimed to elucidate the hepatoprotective mechanisms of XYS bioactive compounds against drug-induced liver injury (DILI) using an integrative network pharmacology and experimental approach. Methods: Network pharmacology was integrated with transcriptomic analysis and machine learning to identify hub targets. Molecular docking was used to predict interaction patterns and relative binding energies between compounds and target proteins. Hepatoprotective effects were validated in an acetaminophen (APAP)-induced hepatotoxicity model in HepG2 cells by measuring cytotoxicity markers and hub gene expression. Results: Network pharmacology identified 125 bioactive compounds with 121 targets overlapping DILI pathogenesis. Machine learning identified five hub genes: ALOX5, CYP1A2, CYP3A4, F3, and PDE5A. Molecular docking showed favorable binding affinities (-5.02 to -8.66 kcal/mol) for four compounds: Beta-sitosterol, kaempferol, luteolin, and quercetin. All four compounds attenuated APAP-induced hepatotoxicity and modulated hub gene expression, with beta-sitosterol showing the most comprehensive hepatoprotective profile. Conclusions: The bioactive compounds of XYS may contribute to hepatoprotection against DILI through multi-target regulation involving the ALOX5-LTB4 inflammatory axis, CYP-mediated metabolic regulation, F3-associated procoagulant signaling, and PDE5A/cGMP-related signaling.en
DOIhttps://doi.org/10.5812/ijpr-171015en
URIhttps://brieflands.com/journals/ijpr/articles/171015en
KeywordHepatoprotectionen
KeywordMachine Learningen
KeywordMulti-target Mechanismsen
KeywordNetwork Pharmacologyen
KeywordPhytochemicalsen
KeywordXiao-Yao-Sanen
PublisherBrieflandsen
TitleIntegrative Network Pharmacology and Experimental Validation Reveal Multi-target Hepatoprotective Mechanisms of Bioactive Compounds from Xiao-Yao-Sanen
TypeResearch Articleen

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