Investigating therapeutic effects and the underlying mechanisms of jia-wei-yin-chen-hao-tang in non-alcoholic fatty liver disease based on network pharmacology analysis and experimental validation.

ETHNOPHARMACOLOGICAL RELEVANCE: Yin-Chen-Hao-Tang (YCHT) was initially recorded in the Chinese medicine classic Shang Han Lun, which was written by Zhang Zhongjing, and it has long been used orally as a classic formula. Jia-Wei-Yin-Chen-Hao-Tang (JWYCHT) comprises YCHT added with the widely used Chinese Yi medicine Liangtoumao (Incarvillea arguta (Royle) Royle). JWYCHT has become a promising therapeutic option for several liver and gallbladder disorders, including nonalcoholic fatty liver disease (NAFLD), among the Chinese Yi population. However, its specific mechanism of action remains poorly understood.

AIM OF THE STUDY: This study aimed to explore the therapeutic effects and underlying mechanism of action of JWYCHT through network pharmacological analysis and experimental validation.

METHODS: NAFLD-related transcriptome data and target genes associated with JWYCHT were retrieved from public databases. Initially, candidate genes were obtained by overlapping differentially expressed genes (DEGs) from NAFLD and control samples in GSE63067 with the drug target genes. Subsequently, candidate biomarkers were further obtained using the algorithm in cytoHubba plug-in. Biomarkers were further identified via receiver operating characteristic (ROC) analysis and expression validation, and enrichment analysis was executed to explore the potential mechanisms of NAFLD biomarkers. HPLC and UPLC/QE-HFX were used to determine the chemical components of JWYCHT. Finally, the findings were validated through molecular biology experiments involving a mouse model of NAFLD induced by high-fat diet.

RESULTS: A total of 416 DEGs were acquired, with 51 candidate genes subjected to enrichment analysis. Seven candidate biomarkers, including C-X-C motif chemokine ligand 10 (CXCL10) and caspase-1 (CASP1), were identified from an algorithm in cytoHubba plug-in. ROC analysis confirmed CASP1 and CXCL10 as viable biomarkers, with the latter (odds ratio = 1.305, p < 0.05) emerging as a risk factor for NAFLD. 41 compounds were identified by UPLC/QE-HFX in JWYCHT. The HPLC results demonstrated distinct variations in chemical profiles between JWYCHT, YCHT, and their component medicinal herbs. A series of molecular experiments revealed that JWYCHT effectively treated NAFLD in the mouse model. It can reduce the expressions of CXCL10 and CASP1, improve liver function biochemical indicators and histopathological changes, and alleviate inflammation and lipid deposition.

CONCLUSION: Bioinformatics and experimental validation revealed that JWYCHT can alleviate hepatic inflammation and lipid deposition in NAFLD by reducing the expressions of CXCL10 and CASP1. Our research results offer an experimental foundation for the therapeutic efficacy and mechanism of JWYCHT and guidance for its clinical application.