Exploration of the synergistic mechanism of Gegen Qinlian Decoction on Wnt signaling pathway based on an integrated strategy of network pharmacology and transcriptomics

Network pharmacology study indicated that Puerarin (PUE), wogonin (WOG), berberine (BER) and glycyrrhetinic acid (GLY) were the active components in GQD and performed synergistic effect on the targets of the Wnt signaling pathway. Pharmacologic research revealed that WOG, BER, GLY performed inhibited effect on SW480 cells and PUE only exhibited effective antitumor activity when combined with GLY. RNA-seq were used to discover synergistic mechanism of PUE-GLY and CTNNB1, CCND1, SMAD4 were identified as synergistic targets inhibited by PUE-GLY. Moreover, mechanism study explored PUE-GLY could affect the Wnt signaling pathway by up-regulated GSK3B and down-regulated CTNNB1 synergistically. Importantly, it showed that GLY could effectively increase the intracellular content of PUE based on HPLC/MS analysis, and this process was achieved by GLY through influencing the targets of membrane’s pathway, such as cell adhesion molecules, focal adhesion and tight junction. Our study revealed multi-target mechanism of GLY, down-regulated CTNNB1 as the material for efficacy and intervene membrane protein (CDH1, CADM1, ITGB2, ICAM1) as courier in the formulae. Moreover, the mechanism of synergistic antitumor action of Puerariae Lobatae Radix (Gegen-Monarch) and Glycyrrhiza Radix et Rhizoma (Gancao-Assistant) on Wnt signaling pathway was explored systematically. Examination of different expression of gene in SW480 cells.