Abstract:

The potential action mechanism of aurantii fructus immaturus and magnoliae officinalis cortex in the treatment of slow transit constipation (STC) was investigated via network pharmacology and metabolomics.The chemical ingredients and targets of aurantii fructus immaturus and magnoliae officinalis cortex were obtained using the traditional Chinese medicine systems pharmacology database and analysis platform. The disease prediction targets of STC were collected through the GeneCards, OMIM, and DisGeNET databases. The intersection targets of ingredients and diseases were obtained using Venn diagrams. The STRING database was used to construct the protein-protein interaction network. The Cytoscape 3.8.0 software was used to calculate and screen the key targets, and then the network diagram of TCM-ingredient targets was plotted. The gene ontology(GO) functional enrichment analysis and Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment analysis of the intersection targets were performed using the Metascape database. A loperamide-induced STC mouse model was used in the study. After intragastric administration of aurantii fructus immaturus and magnoliae officinalis cortex, GC/TOF-MS-based untargeted metabolomics of cecal contents was performed to analyze differential metabolites. A total of 24 active ingredients and 106 intersection targets were obtained. The key targets with higher degree values included AKT1, TNF, TP53, IL6, CASP3, and JUN. GO analysis revealed that the possible processes were cellular response to nitrogen compound, cellular response to lipid, positive regulation of protein phosphorylation, regulation of inflammatory response, regulation of ion transport, etc. KEGG analysis revealed the pathways involved in cancer, the PI3K-Akt signaling pathway, the calcium signaling pathway, serotonergic synapse, etc. In addition, 21 differential metabolites were found via untargeted metabolomics, including the Akt-associated metabolites nicotinic acid, fructose, and protocatechuic acid. The results suggested that aurantii fructus immaturus and magnoliae officinalis cortex exerted therapeutic effects on STC via multi-ingredient, multi-target and multi-pathway mechanisms, thereby providing ideas and a theoretical basis for future basic research. The active ingredients naringenin and lignan, as well as the key target Akt and its related metabolites, deserves special attention.

Key words: Aurantii Fructus Immaturus, Magnoliae Officinalis Cortex, slow transit constipation, network pharmacology, metabolomics