Effect of asiatic acid on sevoflurane-induced apoptosis in hippocampal neurons HT-22 cells by regulating the PI3K/AKT signaling pathway
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.
Asiatic acidPhosphatidylinositol 3-kinase/protein kinase B signaling pathwayHippocampal neuronsCell viabilityApoptosis
NETL
NSTL
万方数据
