Identification of the Active Compound of Liu Wei Di Huang Wan for Treatment of Gestational Diabetes Mellitus via Network Pharmacology and Molecular Docking

Abstract

Liu Wei Di Huang Wan (LWDHW) is a well-known Chinese herbal compound, which has been prescribed for the treatment of gestational diabetes mellitus (GDM). We sought to clarify the potential therapeutic effects of LWDHW against GDM. Differentially expressed genes (DEGs) in GDM were firstly identified from the Gene Expression Omnibus (GEO) database. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses were performed to reveal the biological functions of the DEGs. Subsequently, the LWDHW-compound-target network was constructed based on public databases to identify the relationship between the active components in LWDHW and the corresponding targets. Furthermore, gene functional analysis and protein-protein interaction (PPI) network construction were applied to investigate the function of potential targets and to evaluate hub genes. Finally, molecular docking was used to verify the binding activities between active ingredients and hub targets. Thirteen active components and 39 corresponding therapeutic target genes were obtained via network pharmacology analysis. The enrichment analysis demonstrated that the anti-GDM effect of LWDHW included oxidoreductase activity, involvement in renal system process, and regulation of blood pressure, which may be achieved through regulation of serotonergic synapses, vascular smooth muscle contraction, and neuroactive ligand-receptor interaction pathways. Additionally, molecular docking revealed that the main active component, Mu Dan Pi, exhibited the best affinity for proteins encoded by hub genes. This study applied network pharmacology analysis and molecular docking to display the multicomponent and multitarget characteristics of LWDHW in the treatment of GDM. Our findings provide novel insights into the pathogenesis of GDM and the therapeutic mechanisms of LWDHW against GDM.

Figure 1

(a) Heatmap of differentially expressed genes (DEGs) from the microarray analysis. The orange group is the gestational diabetes mellitus (GDM) group, while the blue group is the control group. The upregulated genes are shown in red, and the downregulated genes are shown in blue. (b) DEGs obtained from the microarray. The red points represent upregulated genes, and blue points represent downregulated genes. Gray points represent genes without a significant difference in expression.

Figure 2

Enrichment analysis of the differentially expressed genes of gestational diabetes mellitus (GDM). (a) Bubble plot of the top 20 enriched biological process (BP). (b) Cellular component (CC) and (c) molecular function (MF) terms revealed by Gene Ontology (GO) functional enrichment analysis of differentially expressed genes (DEGs) in GDM. (d) Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis. (e) Bar plot of the top 20 enriched GO terms. The size of the bubble indicates the gene count, while colors indicate the significance of enrichment.

Figure 3

(a) LWDHW-compound network. (b) LWDHW-compound–target network. The purple triangle represents the LWDHW formulation. The brown diamonds represent the herbs, and the blue squares indicate the constituent compounds. The blue squares with the red frames indicate the main ingredients. Pink circles represent the targets.

Figure 4

Enrichment analysis of gestational diabetes mellitus- (GDM-) related target genes. (a) Bubble plot of the top 20 enriched biological process (BP), (b) cellular component (CC), and (c) molecular function (MF) categories. (d) Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis. (e) Venn diagram of the intersection of differentially expressed genes (DEGs) of GDM and LWDHW targets. (f) Bar plot of the top 20 enriched Gene Ontology (GO) terms. The size of the bubble indicates the gene count, while colors indicate enrichment significance.

Figure 5

(a) hsa04726: serotonergic synapse pathway. (b) hsa04270: vascular smooth muscle contraction pathway. (c) hsa04080: neuroactive ligand–receptor interaction pathway. Red represents the critical genes involved in the pathways.

Figure 6

Protein–protein interaction (PPI) network of gestational diabetes mellitus (GDM) target genes. (a) PPI network, (b) network analyzer network, (c) top 10 hub genes, and (d) top 20 hub genes. The size of the circle represents the significance. From blue to red represents the different log₂ fold change, the thickness of the connection represents the combined score.

Figure 7

LWDHW-main active compound–GDM-target-signaling pathway network. Brown diamonds represent herbs, blue squares represent ingredients, pink circles represent target genes, and red hexagons represent pathways. The pink circles with red frames represent the pathway-related targets.

Figure 8

Molecular docking models of LWDHW binding to the proteins encoded by the top three hub target genes. (a) ADORA2A, (b) CNR1, (c) AGT, (d) catechin, (e) aristolone, (f) mairin, (g) ADORA2A and aristolone, (h) ADORA2A and mairin, (i) AGT and aristolone, (j) CNR1 and catechin, (k) ADORA2A and aristolone, (l) ADORA2A and mairin, (m) AGT and aristolone, and (n) CNR1 and catechin. Hydrogen bonded atoms in the receptor or atoms in close contact with atoms in the ligand are shown as spheres, and fragments of the secondary structure are shown as sequences of three or more residues interacting with the ligand in the receptor.