Salidroside alleviates TNF-α-induced endothelial inflammatory injury by modulating NF-κB/NLRP3 inflammasome-related signaling: an integrated network pharmacology and experimental study.

BACKGROUND

Kawasaki disease (KD) is an acute febrile vasculitis in children, and vascular endothelial injury is a central event in the development of coronary artery lesions. Salidroside (SAL), a natural active compound extracted from Rhodiola rosea, has anti-inflammatory, anti-apoptotic, and immunomodulatory properties. However, its mechanism in KD-related vascular inflammatory injury remains unclear. This study aimed to investigate the protective effects and potential mechanisms of SAL in endothelial injury relevant to KD.

METHODS

Potential targets of SAL were predicted using reverse virtual screening and SwissTargetPrediction. KD-related targets were collected from the GeneCards and DisGeNET databases. Overlapping targets were subjected to Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), and protein-protein interaction analyses. Key targets were further evaluated by molecular docking. Human coronary artery endothelial cells (HCAECs) were stimulated with tumor necrosis factor-α (TNF-α) to establish an endothelial inflammatory injury model. RNA-sequencing (RNA-seq) was performed to identify differentially expressed genes after SAL treatment. The protective effects of SAL were then assessed by Cell Counting Kit-8 assay, wound-healing assay, Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) flow cytometry, reverse transcription quantitative polymerase chain reaction, and western blotting.

RESULTS

Network pharmacology identified 226 potential SAL targets and 2,686 KD-related targets, with 56 overlapping targets. KEGG enrichment and RNA-seq indicated that SAL may regulate a broader inflammatory network, particularly interleukin (IL)-17 and TNF signaling pathways. Molecular docking showed stable binding of SAL to matrix metalloproteinase-9 (MMP-9). In TNF-α-stimulated HCAECs, SAL significantly improved cell viability, inhibited excessive migration, and reduced inflammatory cell death. SAL also decreased the messenger RNA (mRNA) expression of IL-1β, IL-6, MMP-9, interleukin-17 receptor A (IL-17RA), intercellular adhesion molecule-1 (ICAM-1), and NOD-like receptor family pyrin domain containing 3 (NLRP3), and reduced the protein levels of IL-6, phosphorylated nuclear factor kappa B (NF-κB) p65, NLRP3, gasdermin D (GSDMD), and Caspase-1 (P<0.05).

CONCLUSIONS

SAL alleviates TNF-α-induced endothelial inflammatory injury in HCAECs. Its protective effects are associated with suppression of inflammatory signaling, inhibition of abnormal migration and cell death, and attenuation of NF-κB/NLRP3 inflammasome-related molecules. These findings provide preliminary experimental evidence for further investigation of SAL in KD-related vascular inflammation.