A Combined Phytochemistry and Network Pharmacology Approach to Reveal the Effective Substances and Mechanisms of Wei-Fu-Chun Tablet in the Treatment of Precancerous Lesions of Gastric Cancer

Abstract

Wei-Fu-Chun (WFC) tablet is a commercial medicinal product approved by China Food and Drug Administration, which is made of Panax ginseng C.A.Mey., Citrus aurantium L., and Isodon amethystoides (Benth.). WFC has been popularly used for the treatment of precancerous lesions of gastric cancer (PLGC) in clinical practice. In this study, a UHPLC-ESI-Q-TOF/MS method in both positive and negative ion mode was employed to rapidly survey the major constituents of WFC. 178 compounds including diterpenoids, triterpenes, sesquiterpenes, flavonoids, saponins, phenylpropanoids, lignans, coumarins, organic acids, fatty acids, quinones, and sterols, were identified by comparing their retention times, accurate mass within 5 ppm error, and MS fragmentation ions. In addition, 77 absorbed parent molecules and nine metabolites in rat serum were rapidly characterized by UHPLC-ESI-Q-TOF/MS. The network pharmacology method was used to predict the active components, corresponding therapeutic targets, and related pathways of WFC in the treatment of PLGC. Based on the main compounds in WFC and their metabolites in rat plasma and existing databases, 13 active components, 48 therapeutic targets, and 61 pathways were found to treat PLGC. The results of PLGC experiment in rats showed that WFC could improve the weight of PLGC rats and the histopathological changes of gastric mucosa partly by inhibiting Mitogen-activated protein kinase (MAPK) signaling pathway to increase pepsin secretion. This study offers an applicable approach to identify chemical components, absorbed compounds, and metabolic compounds in WFC, and provides a method to explore bioactive ingredients and action mechanisms of WFC.

Keywords: UHPLC-ESI-Q-TOF/MS; Wei-Fu-Chun tablet; effective substances and mechanism; network pharmacology; precancerous lesions of gastric cancer.

FIGURE 1

Representative base peak intensity (BPI) chromatograms of WFC in positive mode (A) and in negative mode (B) and total ions chromatogram (TIC) in positive mode (C) and in negative mode (D) by UHPLC-ESI-Q-TOF/MS.

FIGURE 2

Representative base peak intensity (BPI) chromatograms of absorbed compounds after oral administration of WFC in negative mode (A) and in positive mode (B) by UHPLC-ESI-Q-TOF/MS; Representative base peak intensity (BPI) chromatograms of blank serum in negative mode (C) and in positive mode (D) by UHPLC-ESI-Q-TOF/MS.

FIGURE 3

Biological processes regulated by WFC to treat PLGC. The count of each biological process was shown on the right side of the bar. The p-value of each biological process was less than 0.05. FDR of each biological process was less than 0.05.

FIGURE 4

Pharmacology-network of the “origins-components-targets-pathways” regulated by WFC. The yellow rectangles represent Chinese herbal medicines, while the purple rhombuses represent active components of WFC. The blue ellipse represents metabolites of ginsenoside Rg1. The red rectangles represent target proteins, and the green ellipses represent pathways in Table 6.

FIGURE 5

Pharmacodynamic effect of WFC on PLGC models. (A) The effect of WFC on the weight change of rats with PLGC. (B) Pepsin activity changes in gastric tissue of rats in different groups. (C) Pathological changes of gastric tissue of in different groups of rats (H&E: 100×). *p < 0.05 compared with the normal group; ^#p < 0.05 compared with the model group.

FIGURE 6

Gene expression related to MAPK pathways in gastric tissues of different groups of rats. *p < 0.05 compared with the normal group; ^#p < 0.05 compared with the model group.