Network Pharmacology and Molecular Docking: Exploring the Mechanism of Peppermint in Mastitis Prevention and Treatment in Dairy Cows

Abstract

In order to elucidate the active ingredients, potential targets, and mechanisms of action of peppermint in treating bovine mastitis, this study utilized network pharmacology analysis and molecular docking to conduct an exploratory, prospective investigation. Using the Traditional Chinese Medicine Systems Pharmacology (TCMSP) database, all compounds and targets of peppermint were retrieved. After removing duplicates, a total of 133 compounds and 272 targets were obtained. Targets were then standardized to gene names using the UniProt database to construct a drug-component-target network. A total of 183 disease targets related to bovine mastitis were retrieved from the GeneCards database. We obtained 28 cross targets of peppermint targets and bovine mastitis targets, and constructed a protein-protein interaction (PPI) network using the STRING database. A visual network was built using Cytoscape 3.10.0 software, and seven core targets were analyzed and obtained. GO and KEGG pathway enrichment analysis was performed using the Metascape database. Molecular docking was conducted using AutoDockTools-1.5.6 software on some small-molecule compounds and the seven targets to evaluate the stability of binding between peppermint and core targets. Apigenin, luteolin, and ursolic acid are the three main components in peppermint. Core targets (TNF, IL-6, STAT-3, IL-1β, FGF-2, IFNG, and ESR-1) were selected based on the PPI network. The enrichment analysis suggested that the major signaling pathways in network pharmacology may include AGEs-RAGE, IL-17, NF-κB, TLRs, HIF-1, TGF-β, PI3K-Akt, and MAPK. The molecular docking results showed that one of the main components of mint, ursolic acid, exhibited good binding activity with all core targets of bovine mastitis. Other constituents also produced favorable binding with some core targets. This study elucidates the mechanisms of mint in treating bovine mastitis, providing data to support the potential development of new therapies for bovine mastitis using mint and its constituents.

Keywords: action mechanism; bovine mastitis; mint; molecular docking; network pharmacology.

Figure 1

The figure shows the entire content and process of this study, including screening of peppermint components, screening of disease targets, determination of core targets, GO and KEGG enrichment analysis, and molecular docking.

Figure 2

Mint ingredient–target point network diagram. Trapezoids represent the components of mint, while rectangles denote the corresponding targets for each component. Mint components are indicated by MOL IDs, and each edge signifies an interaction, with color gradients from dark to light representing the degree values.

Figure 3

Potential target analysis of mint in treating mastitis in dairy cows. (A) Venn diagram of potential targets for the mint treatment of bovine mastitis. (B) In the visualized interaction network, orange diamonds represent the core targets, while the remaining circles represent other targets. The shade of color indicates the Degree value, with darker colors signifying a higher Degree. (C) The Betweenness unDir values of the core target are displayed, all of which are greater than the threshold (12.230769230769234). (D) The Closeness unDir values of the core target are displayed, all of which are greater than the threshold (0.027597822814210176). (E) The Degree values of the core target are displayed, all of which are greater than the threshold (12.76923076923077).

Figure 4

The relationship between 7 core objectives and mint components. Purple circles represent the components of mint, colored squares represent the core targets, and straight lines show the connections between them.

Figure 5

GO and KEGG analysis results. (A) The displayed terms are the top ranked biological processes, cellular components, and molecular functions in GO analysis, ranked according to “−log10 (p-value).” (B) Partial KEGG signaling pathway entries, and the X-axis represents the enrichment score.

Figure 6

Molecular docking results of peppermint components with core targets. The image in the upper left corner of each region is a macro 3D visualization, the lower left corner shows the details of the 3D visualization, and the 2D visualization is on the right side. (A) Docking ursolic acid and ESR–1; (B) Docking acacetin and FGF–2; (C) Docking apigenin and FGF–2; (D) Docking acacetin and IFNG; (E) Docking luteolin and FGF–2; (F) Docking naringenin and FGF–2; (G) Docking ursolic acid and IL–1β; (H) Docking ursolic acid and IL–6; (I) Docking ursolic acid and STAT–3.