Identification of in vitro-in vivo components of Caoguo using accelerated solvent extraction combined with gas chromatography-mass spectrometry integrated with network pharmacology on indigestion

BACKGROUND: Caoguo (Tsaoko Fructus), a traditional Chinese medicine, is widely used as medicine and dietary spices. Volatile components are among its important bioactive constituents used to treatment of abdominal distension and pain, but the mechanism is not clear up to now. The purpose of this study was to develop a simple, sensitive, and accurate method to analyze and identify components of Caoguo in vitro and in vivo, and further investigate the therapeutic mechanism of Caoguo on indigestion using network pharmacology. METHODS: Caoguo were extracted by accelerated solvent extraction (ASE) and n-hexane:ethyl acetate (1:1, v/v) was selected as the extraction solvent. Gas chromatography-mass spectrometry (GC-MS) was adopted to analyze and identify the volatile components in vitro and in vivo. Network pharmacology including protein-protein network construction, Gene Ontology (GO) enrichment and pathway enrichment analysis and component-target-pathway network construction was applied. RESULTS: By comparing the retention times and mass spectrometry data, as well as retrieving the reference literature, a total of 169 components were tentatively identified in Caoguo extract and 43 components were identified in rats plasma samples for the first time. The results of network pharmacology analysis indicated that the potential mechanism was mainly associated with regulation of lipolysis in adipocytes and serotonergic synapse signaling pathway, which might be responsible for the effect of indigestion. CONCLUSIONS: Caoguo was first extracted by ASE and the volatile chemical components in vivo were first identified by GC-MS. Coupled with network pharmacology analysis, a network of component-target-pathway was constructed to reveal the possible mechanism of Caoguo in treatment of indigestion. This study provided a new reference method for the extraction and analysis of Caoguo, laid a chemical basis for in-depth studies on pharmacodynamics and pharmacology, and revealed an updated understanding of the therapeutic effects of Caoguo on indigestion.