Integrated network pharmacology, molecular simulations, biophysical validation, and experimental validation to reveal the pharmacological effects and targets of Senkyunolide A against inflammation and oxidative stress

Inflammatory response and oxidative stress interact with each other and are involved in the pathogenesis of various chronic diseases. Senkyunolide A (SenA) is a phthalide compound isolated from the traditional Chinese medicine Chuanxiong Rhizoma (Ligusticum chuanxiong Hort.). At present, the anti‑inflammatory and anti‑oxidative stress effects of SenA remain unclear. In this study, we adopted an integrated strategy combining network pharmacology, bioinformatics analysis, molecular docking, molecular dynamics simulation, bio‑layer interferometry (BLI), and in vitro experiments to explore the anti‑inflammatory and anti‑oxidative stress effects and potential targets of SenA. Through network pharmacology and bioinformatics analysis, we identified four core target genes (Il1b, Ptgs2, Nos2, and Hmox1) of SenA against LPS‑induced inflammation in RAW264.7 cells. Direct binding of SenA to IL‑1β and PTGS2 was confirmed by molecular docking, molecular dynamics simulation, and BLI assays. In vitro experiments showed that SenA pretreatment effectively inhibited LPS‑induced inflammatory response and oxidative stress in RAW264.7 cells, as evidenced by reduced expression of pro‑inflammatory cytokines (TNF‑α, IL‑6, and IL‑1β), decreased levels of NO, ROS, and MDA, increased GSH levels, and alleviated cell swelling and mitochondrial damage. In addition, SenA pretreatment downregulated the mRNA expression levels of the core target genes Il1b, Ptgs2, Nos2, and Hmox1. In conclusion, our findings demonstrate that SenA exerts significant anti‑inflammatory and anti‑oxidative stress effects and may serve as a candidate compound for the treatment of inflammation‑related diseases.