Study on hepatoprotective mechanisms of Veratrilla baillonii based on network pharmacology

Abstract

The purpose of this study was to predict the active components, targets and signaling pathways of Veratrilla baillonii for the prevention and treatment of non-alcoholic liver diseases, preliminarily verify the active components and related targets through cell experiments, and elucidate the mechanism of V. baillonii on liver protection. The candidate active components of V. baillonii were screened by searching Chinese medicine ingredients and Chinese medicine pharmacology database and analysis platform, combined with the pharmacokinetic parameters(oral availability and drug-like principle); the target of candidate active ingredients were predicted by protein database, and the target of disease related to non-alcoholic liver disease was predicted. Cytoscape software was used to construct the network of "active component-target-disease", and the protein interaction network was constructed through the STRING database to infer the core target. GO annotation analysis, KEGG pathway analysis and enrichment analysis were conducted through DAVID bioinformatics annotation database. Finally, the core target and pathway of V. baillonii were preliminarily verified by the experimental model of H_2O_2-induced liver cell damage intervened by V. baillonii water extract(WVBF). The cell viability was detected by MTT assay and real-time unlabeled assay, and the expression of related genes was analyzed by Real-time quantitative polymerase chain reaction(PCR). Firstly, 14 active components were obtained from V. baillonii through network pharmacology. There were 287 potential targets corresponding to the components, 587 targets related to non-alcoholic liver disease, and 13 core targets after the interaction between active ingredient targets and disease targets. Secondly, GO enrichment analysis showed that these genes mainly affected 26 biological processes such as nuclear receptor activity, transcription factor activity, steroid hormone receptor activity, ubi-quitin-like protein ligase binding, protein heterodimerization activity, and transcription cofactor binding. KEGG enrichment analysis showed that PI3 K-AKT signaling pathway, HIF-1 signaling pathway, MAPK signaling pathway, insulin signaling pathway, TNF signaling pathway and some cancer-related pathways were more enriched. Finally, TNF-α and MAPK8 were successfully verified as important targets by hepatocytes in vitro, which suggested that V. baillonii could significantly improve liver damage. TNF-α and MAPK8 were one of the targets. Based on the above results, we systematically predicted the material basis and biological mechanism of V. baillonii through multi-component, multi-target and multi-pathway regulation of nonalcoholic liver disease, and the core targets were successfully verified by cells, providing data basis and scientific basis for the in-depth development of V. baillonii.