Exploring the targets and molecular mechanism of glycyrrhetinic acid against diabetic nephropathy based on network pharmacology and molecular docking

Figure 1 Graphical abstract summarizing the network pharmacological predictions and experimental validations. GA: Glycyrrhetinic acid; PPI: Protein-protein interaction; GO: Gene Ontology; KEGG: Kyoto Encyclopedia of Genes and Genomes.

Figure 2 The structure of drug active ingredient. A: Beta-glycyrrhetinic acid; B: Alpha-glycyrrhetinic acid.

Figure 3 Targets and protein-protein interaction network enrichment analysis. A: Drug targets and disease genes; B: Intersection of drug and disease targets; C: Protein-protein interaction (PPI) network diagram of intersection genes between glycyrrhetinic acid (GA) and the disease; D: Utilization of Cytoscape to construct a PPI network comprising 186 common targets and 2 core targets for potential disease treatment; E: Analysis results of selected core targets and GA’s active components. Each node’s color denotes its degree.

Figure 4 Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analysis. A: Results of Gene Ontology enrichment analysis; B: Enrichment outcomes for Kyoto Encyclopedia of Genes and Genomes pathways; C: Expression results of enriched core genes within the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (AKT) signaling pathway; D: Correlation between the PI3K/AKT signaling pathway and the cell cycle.

Figure 5 Molecular docking. A: Demonstration of docking outcomes between glycyrrhetinic acid (GA) components and core target molecules; B: Presentation of docking results and binding energies between GA components and core target molecules; C: Heat map display of molecular docking binding energy results. GA: Glycyrrhetinic acid.

Figure 6 Determination of IC₅₀, cell cycle, and apoptosis. A: Impact of glycyrrhetinic acid (GA) on the IC₅₀ of HK-2 cells; B: Influence of GA on the activity of HK-2 cells; C: Effect of GA on the cell cycle of HK-2 cells; D: Effect of GA on apoptosis of HK-2 cells. The sequence from left to right represents the control group, the model group, and the GA group. ^aP < 0.05, ^bP < 0.01, ^cP < 0.001, ^dP < 0.0001. GA: Glycyrrhetinic acid; CON: Control; MOD: Model.

Figure 7 Impact of glycyrrhetinic acid on signal proteins in the phosphatidylinositol 3-kinase/protein kinase B signaling pathway. A: Expression levels of insulin receptor and glycogen synthase kinase-3 proteins; B: Protein levels of phosphatidylinositol 3-kinase (PI3K)/p-PI3K and protein kinase B (AKT)/p-AKT. The sequence from left to right represents the control group, the model group, and the glycyrrhetinic acid group. ^aP < 0.05, ^bP < 0.01, ^cP < 0.001. GA: Glycyrrhetinic acid; CON: Control; MOD: Model; GSK3: Glycogen synthase kinase-3; INSR: Insulin receptor; AKT: Protein kinase B; PI3K: Phosphatidylinositol 3-kinase.

Figure 8 Diagram of the effect of glycyrrhetinic acid on the PI3K/AKT signaling pathway. GSK3: Glycogen synthase kinase-3; INSR: Insulin receptor.