Use of Network Pharmacology and Molecular Docking Technology to Analyze the Mechanism of Action of Velvet Antler in the Treatment of Postmenopausal Osteoporosis

Abstract

Deer velvet antlers are the young horns of male deer that are not ossified and densely overgrown. Velvet antler and its preparations have been widely used in the treatment of postmenopausal osteoporosis (PMOP) in recent years, although its mechanism of action in the human body remains unclear. To screen the effective ingredients and targets of velvet antler in the treatment of PMOP using network pharmacology and to explore the potential mechanisms of velvet antler action in such treatments, we screened the active ingredients and targets of velvet antler in the BATMAN-TCM database. We also screened the relevant targets of PMOP in the GeneCards and OMIM databases and then compared the targets at the intersection of both velvet antler and PMOP. We used Cytoscape 3.7.2 software to construct a network diagram of "disease-drug-components-targets" and a protein-protein interaction (PPI) network through the STRING database and screened out the core targets; the R language was then used to analyze the shared targets between antler and PMOP for GO-enrichment analysis and KEGG pathway-annotation analysis. Furthermore, we used the professional software Maestro 11.1 to verify the predictive analysis based on network pharmacology. Hematoxylin-eosin (H&E) staining and micro-CT were used to observe the changes in trabecular bone tissue, further confirming the results of network pharmacological analysis. The potentially effective components of velvet antler principally include 17β-E2, adenosine triphosphate, and oestrone. These components act on key target genes such as AKT1, IL6, MAPK3, TP53, EGFR, SRC, and TNF and regulate the PI3K/Akt-signaling and MAPK-signaling pathways. These molecules participate in a series of processes such as cellular differentiation, apoptosis, metabolism, and inflammation and can ultimately be used to treat PMOP; they reflect the overall regulation, network regulation, and protein interactions.

Figure 1

Venn diagram of velvet antler and postmenopausal osteoporosis (PMOP) targets. Light blue represents the PMOP targets, and red represents the velvet targets. The intersection of the two colors represents a total of 233 potential targets for velvet antler treatment of PMOP.

Figure 2

The “disease-drug-component-target” regulatory network of velvet antler in the treatment of postmenopausal osteoporosis. The triangle represents the target, the diamond represents the active ingredients of velvet antler, the regular octagon represents “velvet drugs,” and the hexagon represents postmenopausal osteoporotic disease. The straight lines connecting each node indicate the presence of regulatory relationship between the nodes.

Figure 3

PPI network. The color and size of the nodes reflect the degree value. The larger and darker the nodes in the graph, the greater the DC value.

Figure 4

Analysis diagram of GO functional enrichment. The ordinate represents GO function items, the abscissa represents the proportion of genes, the size of the dots represents the number of enriched genes, and the color of the dots represents the range of different P values.

Figure 5

KEGG analysis diagram of key targets. The ordinate represents the name of the KEGG signal pathway, the abscissa represents the proportion of genes, the size of the dots represents the number of enriched genes, and the color of the dots represents the range of different P values.

Figure 6

KEGG- and PI3K/Akt-signaling pathway: □ represents gene products; ○ represents compounds; red represents the key targets of deer antler; ⟶ represents intermolecular interactions or relationships.

Figure 7

The molecular docking model of the top three active ingredients in velvet antler and the eleven core targets. In the 3D diagram, the red dotted line indicates the hydrogen bonds and related residues connected to each other, and the text represents the name of the residue. (a) AKT1 and 17-beta-estradiol; (b) AKT1 and oestrone; (c) AKT1 and adenosine triphosphate; (d) IL6 and adenosine triphosphate; (e) EGFR and 17-beta-estradiol; (f) EGFR and oestrone; (g) ESR1 and 17-beta-estradiol; (h) ESR1 and oestrone; (i) SRC and 17-beta-estradiol; (j) SRC and oestrone; (k) SRC and adenosine triphosphate; (l) STAT3 and 17-beta-estradiol; (m) STAT3 and oestrone; (n) STAT3 and adenosine triphosphate; (o) TNF and adenosine triphosphate; (p) MAPK1 and 17-beta-estradiol; (q) MAPK1 and oestrone; (r) MAPK1 and adenosine triphosphate; (s) JUN and 17-beta-estradiol; (t) JUN and oestrone; (u) JUN and adenosine triphosphate.

Figure 8

Micro-CT images and morphometric parameters in the distal femur. ^∗p < 0.05; ^∗∗p < 0.01.

Figure 9

Hematoxylin and eosin staining of the distal femur, magnification ×200.

Figure 10

Static trabecular histomorphometry parameters after 3 weeks of antler treatment (^∗p < 0.05; ^∗∗p < 0.01).