Exploring the Effect of Esketamine on IL-1β-Induced Chondrocyte Apoptosis Based on PI3K/AKT/mTOR Pathway
XIN Xuedong¹, BAI Yuguang¹, NIU Yaqing², XIE Yaying¹*
This study aimed to explore the effect of esketamine on IL-1β (interleukin-1β)-induced chondro cyte apoptosis by regulating the PI3K (phosphatidylinositol 3-kinase)/AKT (protein kinase B)/mTOR (mammalian target of rapamycin) signaling pathway. Human chondrocytes were divided into four groups: the Control group, the IL-1β group (10 μg/L IL-1β), the IL-1β+esketamine group (10 μg/L IL-1β combined with 100 μmol/L esketamine), and the IL-1β+esketamine+LY294002 group (10 μg/L IL-1β, 100 μmol/L esketamine, and 25 μmol/L LY294002). Cell proliferation was detected by the CCK-8 assay and EdU staining; cell apoptosis was measured by flow cy tometry and TUNEL staining; intracellular ROS (reactive oxygen species) levels were determined by the DCFH DA method; the contents of TNF-α (tumor necrosis factor-α), IL-6 (interleukin-6), and PGE2 (prostaglandin E2) in the cells were detected by ELISA (enzyme-linked immunosorbent assay); NO (nitric oxide) content was measured by the Griess reaction; the expression levels of apoptosis-related proteins and PI3K/AKT/mTOR pathway-related proteins were determined by Western blot assay. Compared with the Control group, the IL-1β group showed signifi cant decreases in cell viability, EdU-positive cell rate, Bcl-2 protein expression, and the p-PI3K (phosphorylation levels of PI3K), p-AKT (phosphorylation levels of AKT) and p-mTOR (phosphorylation levels of mTOR), while cell apoptosis rate, TUNEL-positive rate, the levels of ROS, TNF-α, IL-6, NO and PGE2, as well as the expression levels of Bax, Cleaved caspase-3 and Cleaved caspase-9 proteins were significantly increased (all P<0.05). Com pared with the IL-1β group, the IL-1β+esketamine group exhibited notable increases in cell viability, EdU-positive cell rate, Bcl-2 protein expression, and the phosphorylation levels of PI3K, AKT and mTOR, whereas cell apop tosis rate, TUNEL-positive rate, the levels of ROS, TNF-α, IL-6, NO and PGE2, and the expression levels of Bax, Cleaved caspase-3 and Cleaved caspase-9 proteins were markedly decreased (all P<0.05). Treatment with the PI3K pathway inhibitor reversed the ameliorative effect of esketamine on IL-1β-induced chondrocyte injury (P<0.05). In summary, esketamine may inhibit IL-1β-induced chondrocyte apoptosis by activating the PI3K/AKT/mTOR signal ing pathway.
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