Network pharmacology, molecular docking and experimental verification reveal the mechanism of Yiguanjian decoction in treating acute liver failure

Abstract

Introduction: The potential targets and action mechanism of Yiguanjian (YGJ) decoction in treating acute liver failure (ALF) were determined using network pharmacology, molecular docking and cell experiments.

Methods: The Pharmacology of Traditional Chinese Medicine Systems (TCMSP) and BATMAN-TCM databases were used to find YGJ decoction targets. The Genecard database was employed to predict targets associated with ALF. Cytascape software was utilised to visualise and select common targets along with establishing a protein–protein interaction (PPI) network. Core targets were screened and putative function was assessed via gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyzes. Molecular docking was performed using AutodockTools, PyMoL and Discovery Studio software to verify the correlation between the YGJ decoction and selected targets. MTT assay was conducted to determine the effect of the YGJ decoction on the viability of Huh-7 human hepatoma cells. The effects of the YGJ decoction on the expression of putative targets in Huh-7 cells were determined via western blot and quantitative real-time polymerase chain reaction analyzes.

Results: Overall, 9153 YGJ decoction and 576 ALF-related targets were obtained, and 469 YGJ decoction and ALF cross targets were obtained. Of these, 20 key targets, including AKT1, estrogen receptor 1 (ESR1), catalase (CAT), interleukin-1β (IL-1β) and discs large homolog 4 (DLG4), were selected from the PPI network. GO function enrichment analysis revealed that these targets were primarily associated with regulating system processes, cell body, oxidoreductase activity and other processes. An enrichment analysis using the KEGG pathway database revealed that the treatment of ALF using the YGJ decoction was primarily associated with the AGE-RAGE, cGMP-PKG and HIF-1 signalling pathways. Molecular docking indicated that quercetin, stigmasterol and β- sitosterol as well as AKT1, ESR1, CAT, IL-1β and DLG4 exhibited good binding affinity. In vitro experiments revealed that the YGJ decoction significantly reversed lipopolysaccharide (LPS)-induced apoptosis (P < 0.01), inhibited LPS-induced increase in IL-1β and ESR1 levels and upregulated LPS-induced decrease in AKT1, CAT and DLG4 levels (P < 0.05).

Conclusions: The YGJ decoction alleviates ALF by regulating multiple targets and its action mechanism may be associated with ameliorating oxidative stress and inflammation.