Melatonin: Multi-Target Mechanism Against Diminished Ovarian Reserve Based on Network Pharmacology

Abstract

Background: Diminished ovarian reserve (DOR) significantly increases the risk of female infertility and contributes to reproductive technology failure. Recently, the role of melatonin in improving ovarian reserve (OR) has attracted widespread attention. However, details on the pharmacological targets and mechanisms of melatonin-improved OR remain unclear.

Objective: A systems pharmacology strategy was proposed to elucidate the potential therapeutic mechanism of melatonin on DOR at the molecular, pathway, and network levels.

Methods: The systems pharmacological approach consisted of target identification, data integration, network construction, bioinformatics analysis, and molecular docking.

Results: From the molecular perspective, 26 potential therapeutic targets were identified. They participate in biological processes related to DOR development, such as reproductive structure development, epithelial cell proliferation, extrinsic apoptotic signaling pathway, PI3K signaling, among others. Eight hub targets (MAPK1, AKT1, EGFR, HRAS, SRC, ESR1, AR, and ALB) were identified. From the pathway level, 17 significant pathways, including the PI3K-Akt signaling pathway and the estrogen signaling pathway, were identified. In addition, the 17 signaling pathways interacted with the 26 potential therapeutic targets to form 4 functional modules. From the network point of view, by regulating five target subnetworks (aging, cell growth and death, development and regeneration, endocrine and immune systems), melatonin could exhibit anti-aging, anti-apoptosis, endocrine, and immune system regulation effects. The molecular docking results showed that melatonin bound well to all hub targets.

Conclusion: This study systematically and intuitively illustrated the possible pharmacological mechanisms of OR improvement by melatonin through anti-aging, anti-apoptosis, endocrine, and immune system regulation effects.

Keywords: biological processes; diminished ovarian reserve (DOR); melatonin; network pharmacology; ovarian reserve (OR); potential therapeutic targets; signaling pathways.

Figure 1

Research workflow diagram.

Figure 2

Melatonin−putative target network. The blue-colored nodes represent the potential targets. The yellow-colored nodes represent the known targets. The red-colored nodes represent the intersection of the potential and known targets.

Figure 3

PPI network related to DOR. The color of the nodes is illustrated from red to cyan in descending order of degree values.

Figure 4

Venn diagram and PPI network of potential therapeutic targets. (A) Venn diagram of intersected targets of melatonin and DOR. (B) PPI network of potential therapeutic targets. The node sizes and colors are illustrated from large to small and orange to green in descending order of degree values.

Figure 5

GO enrichment analysis and the top 5 enriched biological processes. (A) GO enrichment analysis. The top 10 significantly enriched terms of each part. BP, biological process; CC, cell component; MF, molecular function. (B) The top 5 enriched biological processes.

Figure 6

The KEGG pathway analysis of the 26 potential therapeutic targets. (A) The 17 significant pathways. The bubbles’ sizes are indicated from large to small in descending order of the count of the potential targets enriched in the pathways. The bubbles’ colors are indicated from red to blue in descending order of -lg (p-value). (B) Melatonin-targets-pathways network. The width of the line is proportional to the number of connected points. (C) Module analysis of the target-pathway network. The diamond nodes represent the pathways, and the circular nodes represent the targets. The red nodes represent the hub genes obtained from the PPI network of potential therapeutic targets.

Figure 7

Distribution of the potential therapeutic targets on significantly enriched pathways. The red nodes represent key genes, the yellow nodes represent overlapping targets of Melatonin and DOR targets, and the green nodes represent the other targets in estrogen signaling pathway and PI3K-AKT signaling pathway.

Figure 8

The KEGG pathway class analysis and sub-networks in different pathway classes. (A) The pathway class distribution. (B–F) Melatonin’s target sub-networks in different pathway classes. (B) Aging; (C) Cell growth and death; (D) Development and regeneration; (E) Endocrine system; and (F) Immune system. The circular nodes indicate the primary proteins, and the diamond nodes indicate secondary proteins. The pink nodes indicate the common targets of melatonin and DOR; the red nodes indicate the PPI network’s hub genes of potential therapeutic targets; the cyan nodes indicate the melatonin targets, and the deep blue nodes indicate the DOR targets.

Figure 9

Molecular docking of the eight hub targets with Melatonin. (A) The binding poses of MAPK1 complexed with melatonin. (B) The binding poses of AKT1 complexed with melatonin. (C) The binding poses of EGFR complexed with melatonin. (D) The binding poses of HRAS complexed with melatonin. (E) The binding poses of SRC complexed with melatonin. (F) The binding poses of ESR1 complexed with melatonin. (G) The binding poses of AR complexed with melatonin. (H) The binding poses of ALB complexed with melatonin.