Objective  To investigate the effect of asiatic acid (AA) on the apoptosis of HT-22 cells induced by sevoflurane (SEVO) by regulating phosphatidylinositol 3-kinase/protein kinase B (PI3K/AKT) signaling pathway.

Methods  Different concentrations of AA (0, 5, 10, 15, 20, 30 μmol/L) were used to treat HT-22 cells induced by sevoflurane for 24  hours, and CCK-8 was used to detect HT-22 cell viability; HT-22 cells were divided into control group, sevoflurane (SEVO) group, AA low concentration (AA- L, 10 μmol/L) group, AA medium concentration (AA-M, 15 μmol/L) group, AA high concentration (AA-H, 20 μmol/ L) group, and AA high concentration+PI13K pathway inhibitor LY294002 (AA-H+LY294002, 20  μmol/ L AA+5 μmol/ L LY294002) group. Inverted microscopy was applied to observe changes of cell morphology, ELISA was applied to detect the levels of inflammatory factors TNF-α and IL-6, oxidative stress indicators SOD, GSH-Px, and MDA, ROS detection kit was applied to detect ROS levels, TUNEL kit was applied to detect HT- 22 apoptosis, JC-1 method was applied to detect mitochondrial membrane potential, ATP content detection kit was applied to detect ATP content, and Western blot was applied to detect the expressions of Bcl-2, Bax, Caspase-3, p-PI3K, PI3K, p-AKT, and AKT proteins.

Results  Compared with 0 μmol/L, the activity of HT-22 cells treated with 5-30 μmol/ L AA increased in a concentration-dependent manner (P<0.05), concentrations of 10 μmol/ L, 15  μmol/L, and 20 μmol/L of AA were selected for subsequent experiments. Compared with the SEVO group, the levels of TNF-α, IL-6, MDA, ROS, cell apoptosis rate, and expressions of Bax and Caspase-3 proteins in the AA-L, AA-M, and AA-H groups were reduced, the levels of SOD and GSH-Px, red/green JC-1 fluorescence ratio, content of ATP, the expression of Bcl-2 protein, the phosphorylation levels of PI3K and AKT were increased (P<0.05), and were concentration dependent. LY294002 was able to reverse the protective effect of AA on HT-22 cell damage induced by sevoflurane (P<0.05).

Conclusion  AA protects HT-22 cells from damage induced by sevoflurane by activating the PI3K/AKT signaling pathway, which provides a theoretical reference for the development of novel drugs to reduce sevofluran-induced neurotoxicity.

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