α-Cyperone Alleviates LPS-Induced Pyroptosis in Rat Aortic Endothelial Cells via the PI3K/AKT Signaling Pathway

Objective: To investigate the effect and underlying mechanism of alpha-cyperone in inhibiting pyroptosis in rat aortic endothelial cells (RAECs). Methods: Molecular docking technology was used to predict the potential binding affinity of alpha-cyperone to pyroptosis-related proteins. A pyroptosis model was established in RAECs using rat serum containing 10% LPS, with alpha-cyperone administered as a preventive treatment for 9 h. Cell viability and membrane integrity were assessed using propidium iodide (PI) staining and the CCK-8 assay. The release of IL-1 beta and IL-18 was quantified by ELISA. Western blot and RT-qPCR were performed to evaluate the expression levels of NLRP3, ASC, caspase-1 p20, and N-GSDMD. Additionally, RNA sequencing analysis was conducted to identify differentially expressed genes related to pyroptosis in LPS-induced RAECs following alpha-cyperone treatment, and key differential genes were validated by Western blot. Results: Molecular docking analysis reveals that alpha-cyperone exhibits a strong binding affinity to pyroptosis-related targets. alpha-Cyperone significantly improves LPS-induced cell viability (p < 0.001), reduces IL-1 beta and IL-18 release (p < 0.001), and downregulates the mRNA and protein expression of NLRP3, ASC, caspase-1, and GSDMD (p < 0.001). RNA sequencing indicates that alpha-cyperone primarily modulates pyroptosis-related gene expression in RAECs through the PI3K/AKT signaling pathway. Western blot validation further confirmed that alpha-cyperone effectively inhibited the protein expression of phosphorylated and total PI3K and AKT in RAECs (p < 0.001). Conclusions: alpha-Cyperone significantly alleviates morphological damage in the RAEC pyroptosis model, suppresses the release of proinflammatory cytokines IL-1 beta and IL-18, and potentially inhibits NLRP3/caspase-1/GSDMD activation through the PI3K/AKT signaling pathway, thereby attenuating LPS-induced pyroptosis in RAECs.