Integrated Network Pharmacology, Single-Cell Transcriptomics Unveil the Mechanistic Role of Morusin in Aortic Dissection

Abstract

Aortic dissection is a life-threatening cardiovascular emergency with limited pharmacological options. This study focuses on elucidating the multi-target and multi-pathway mechanisms through which morusin mitigates aortic dissection progression, integrating network pharmacology, single-cell transcriptomics and experimental validation. Multi-database analysis identified 281 morusin targets and 1741 ad-related genes, with 84 overlaps. Enrichment analyses highlighted IL-17, HIF-1 and MAPK signalling pathways as potential regulatory hubs. Protein-protein interaction network analysis identified seven key targets, all showing high binding affinity to morusin in molecular docking. Single-cell transcriptomics revealed cell-type-specific dysregulation, notably MAPK8 upregulation in fibroblasts and immune cells. In vitro, morusin dose-dependently inhibited AngII-induced vascular smooth muscle cell proliferation and modulated IL-17 pathway gene expression. In vivo, morusin attenuated aortic dilation and reduced morbidity and mortality in a BAPN-induced AD mouse model. These findings suggest that morusin mitigates AD progression by targeting key inflammatory and apoptotic pathways, supporting its potential as a multi-target therapeutic candidate.

Keywords: IL‐17 signalling pathway; Morusin; aortic dissection; molecular docking; network pharmacology; single‐cell transcriptomics.

FIGURE 1

Research workflow diagram of this study.

FIGURE 2

Identification of morusin‐related targets and AD‐related genes. (A) The 10 representative terms with the lowest p‐value of biological processes (BPs), cellular components (CCs) and molecular functions (MFs) of the GO enrichment analysis of morusin‐related targets (p < 0.05). (B) The 10 significantly enriched KEGG pathways of morusin‐related targets (p < 0.05). (C) The 10 representative terms with the lowest p‐value of BP, CC and MF of the GO enrichment analysis of AD‐related target genes (p < 0.05). (D) The 10 significantly enriched KEGG pathways of AD‐related target genes (p < 0.05).

FIGURE 3

Identification of morusin potential therapeutic pathways. (A) 84 common targets of morusin and AD‐associated targets were visualised by Venn diagram. (B) The 10 representative terms with the lowest p‐value of biological processes (BPs), cellular components (CCs) and molecular functions (MFs) of the GO enrichment analysis of the common targets (p < 0.05). (C) The 10 significantly enriched KEGG pathways of the common targets (p < 0.05). (D) cnet plot of the interactions between the common target genes and KEGG pathways. Red circular nodes represent key molecular targets, while yellow circular nodes denote KEGG pathways. Edges indicate associations between targets and pathways, the size of the nodes represents the quantity of genes. (E) The PPI network of morusin in AD, the screening conditions of hub genes are degree centrality (DC) > 19.976, betweenness centrality (BC) > 74.452 and closeness centrality (CLC) > 0.0065. (F) The position of the core target of morusin in improving AD within the IL‐17 pathway. The red rectangular nodes represent core genes.

FIGURE 4

Molecular docking results of morusin and hub targets. (A) Morusin‐HSP90AA1. (B) Morusin‐MAPK8. (C) Morusin‐MAPK2. (D) Morusin‐PTGS2. (E) Morusin‐MAPK14. (F) Morusin‐NFKB1. (G) Morusin‐CASP3.

FIGURE 5

The hub targets expression in different AD cell types. (A, B) UMAP plots show 22 clusters and 6 cell types in AD patients. (C–I) The hub targets expression in different cell types.

FIGURE 6

Effects of Morusin Intervention on IL‐17 Pathway‐Related mRNA Levels and Cell Viability in HAVSMCs. (A) Effect of morusin on the viability of HAVSMCs. (B) Changes in HSP90AA1 gene expression levels. *p < 0.05. (C) Changes in MAPK8 gene expression levels. (D) Changes in MAPK1 gene expression levels. (E) Changes in MAPK14 gene expression levels. (F) Changes in PTGS2 gene expression levels. ***p < 0.001. (G) Changes in NFKB1 gene expression levels. (H) Changes in CASP3 gene expression levels. **p < 0.01, ***p < 0.001 (n = 4).

FIGURE 7

Supplementation with Morusin attenuates aortic dissection in vivo. (A) Four‐week‐old male mice were administered with 0.25% BAPN (wt/vol) for 30days with or without morusin (40 mg/kg in saline): Vehicle + BAPN, n = 13; and morusin + BAPN, n = 13. (B) Maximal aortic diameter measurement of each group. **p < 10 vehicle + BAPN, n = 10; morusin + BAPN, n = 12. (C) Incidence of aortic dissection for each group. (D) Representative images of haematoxylin and eosin (20X: Scale bar = 50 μm; 40X: Scale bar = 20 μm) and Elastic Van Gieson (20X: Scale bar = 50 μm; 40X: Scale bar = 20 μm) staining in the thoracic aorta of mice.