OBJECTIVE Combining network pharmacology methods and experimental verification to explore the target and potential mechanism of Compound Fresh Dendrobium Granules in treating alcoholic liver disease (ALD).
METHODS With the help of TCMSP, ETCM, BATMAN-TCM databases and literatures, the chemical components of Compound Fresh Dendrobium Granules were searched. The optimal target compounds were screened by Swiss ADME database, and the active ingredient targets were predicted by Swiss Target Prediction database. The targets of ALD-related diseases were obtained from GeneCards, OMIM, DrugBank and DisGeNET databases. The protein-protein interaction (PPI) network analysis and "traditional Chinese medicine (TCM)-active ingredient-target-disease" network diagram were constructed using String database and Cytoscape software. The predicted potential targets were analyzed by GO/KEGG functional enrichment through DAVID database to predict the mechanism of action. A mouse model of alcoholic liver injury was constructed, and the results of network pharmacology enrichment analysis were verified by pathological staining, ELISA, qPCR and Western blot.
RESULTS A total of 73 key active ingredients were obtained by database screening and literatures, and 720 targets were intersected with ALD. 789 BP information, 93 CC information, 204 MF information, and 194 KEGG signaling pathways were obtained by GO annotation analysis based on P < 0.05 and FDR < 0.05, which were mainly involved in metabolic pathways, PI3K/Akt signaling pathway, MAPK signaling pathway, cAMP signaling pathway, calcium signaling pathway, etc. In vivo experiments showed that, compared with the control group, mice in the model group had disorganized hepatocyte cords, indistinct hepatic lobule boundaries, obvious inflammatory cell infiltration and fat vacuoles in hepatocytes, significantly higher serum ALT and AST activity, liver tissue TG, TCHO content, and the gene expression levels of IL-1β, CXCL10 and Srebp-1c, while significantly lower CPT1 gene expression levels. The expression levels of inflammatory pathway proteins p-AKT, p-mTOR and p-p65 proteins were increased. After treatment with Compound Fresh Dendrobium Granules, the inflammatory cell infiltration and fat vacuoles in mouse hepatocytes were reduced, the cellular structure of the liver was normalized, and serum ALT and AST activity, hepatic tissue TG, TCHO content, and the gene expression levels of IL-1β, CXCL10, and Srebp-1c were significantly reduced, while the gene expression levels of CPT1 were significantly elevated. Additionally, the p-AKT, p-mTOR and p-p65 protein expression levels were also significantly reduced.
CONCLUSION Compound Fresh Dendrobium Granules can effectively improve hepatic inflammation and lipid metabolism in mice with alcoholic liver injury, and may inhibit tissue inflammation by regulating the PI3K/Akt signaling pathway. This study preliminarily revealed the mechanisms of Compound Fresh Dendrobium Granules in preventing and treating alcoholic liver disease, which provides a reference for clinical medication and follow-up research.
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