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 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.