Effect of Baicalein on PI3K/Akt Signaling Pathway in HT22 Cell Dementia Model
Objective:To observe effect of baicalein on the expression of PI3K/Akt signaling pathway related factors in HT22 cells induced by Aβ25-35.Methods:The Aβ25-35 at 40 µmol·L-1 was used to intervened in HT22 cells for 24 hours to establish a dementia cell model.Four different concentrations(5,10,20,40 µmol·L-1)of baicalein were used to intervene in dementia cell models for 24 hours.The cell viability was detected by CCK-8 method,and the baicalein concentration with the highest cell viability was selected for subsequent trials.The trial models were divided into the control group,the Aβ25-35 group,and the baicalein group.Flow cytometry and Hoechest 33342 staining were used to detect cell apoptosis;JC-10 method was used to detect mitochondrial membrane potential;DCFH-DA method was used to detect levels of reactive oxygen species;Immunofluorescence was used to detect microtubule associated protein light chain 3(LC3)protein expression;Western blot was used to detect the expression of selective autophagy adaptor protein(p62),autophagy key molecule yeast Atg6 homolog 1(Beclin-1),phosphatidylinositol 3-kinase(PI3K),phosphorylated phosphatidylinositol 3-kinase(p-PI3K),serine/threonine protein kinase(Akt),phosphorylated serine/threonine protein kinase(p-Akt),and mammalian target of rapamycin(mTOR),and compared with the PI3K specific inhibitor LY294002.Results:Finally,a concentration of 10 µmol·L-1 of baicalein was chosen for the subsequent trial.Compared with those in the control group,the apoptosis rate,ROS level,p62 protein expression,and mTOR protein expression were increased in the Aβ25-35 group(P＜0.05),while MMP level,LC3 expression,Beclin-1 protein expression,p-PI3K/PI3K ratio,and p-Akt/Akt ratio were decreased(P＜0.05);Compared with those in the Aβ25-35 group,the cell apoptosis rate,ROS level,p62 protein expression,and mTOR protein expression in the baicalein group were decreased(P＜0.05),while MMP levels,LC3 expression,Beclin-1 protein expression,p-PI3K/PI3K ratio,and p-Akt/Akt ratio were raised(P＜0.05).Compared with those in the baicalein group,the addition of PI3K inhibitor LY294002 resulted in a decrease in the ratio of p-PI3K/PI3K and p-Akt/Akt,and an increase in mTOR protein expression(P＜0.05).Conclusion:Baicalein can significantly improve the mitochondrial function of HT22 cells induced by Aβ25-35,and reduce cell apoptosis and oxidative stress.Its anti-dementia effect is related to the activation of the PI3K/AKT signaling pathway and the increase of cellular autophagy levels.
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