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

<b>Objective:</b> To investigate the effect and underlying mechanism of α-cyperone in inhibiting pyroptosis in rat aortic endothelial cells (RAECs). <b>Methods:</b> Molecular docking technology was used to predict the potential binding affinity of α-cyperone to pyroptosis-related proteins. A pyroptosis model was established in RAECs using rat serum containing 10% LPS, with α-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-1β 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 α-cyperone treatment, and key differential genes were validated by Western blot. <b>Results:</b> Molecular docking analysis reveals that α-cyperone exhibits a strong binding affinity to pyroptosis-related targets. α-Cyperone significantly improves LPS-induced cell viability (<i>p</i> < 0.001), reduces IL-1β and IL-18 release (<i>p</i> < 0.001), and downregulates the mRNA and protein expression of NLRP3, ASC, caspase-1, and GSDMD (<i>p</i> < 0.001). RNA sequencing indicates that α-cyperone primarily modulates pyroptosis-related gene expression in RAECs through the PI3K/AKT signaling pathway. Western blot validation further confirmed that α-cyperone effectively inhibited the protein expression of phosphorylated and total PI3K and AKT in RAECs (<i>p</i> < 0.001). <b>Conclusions:</b> α-Cyperone significantly alleviates morphological damage in the RAEC pyroptosis model, suppresses the release of proinflammatory cytokines IL-1β and IL-18, and potentially inhibits NLRP3/caspase-1/GSDMD activation through the PI3K/AKT signaling pathway, thereby attenuating LPS-induced pyroptosis in RAECs.