The Effect Of Autophagy On Ox-LDL Induced HT-22 Cell Damage

?Objective?The present study was performed to explore whether ox-LDL induced HT-22 neural cell damage was via autophagy impairment and apoptosis enhancement.?Methods?HT-22 cells were treated with different concentration of ox-LDL and then detected the cell viability using Cell Counting Kit-8(CCK-8).Then,Chloroquine(CQ,an inhibitor of autophagy flux)and rapamycin(Rap,an inducer of autophagy)were utilized before ox-LDL to HT-22 cells.CCK-8 was used to detect the cell viability.Flow cytometry and transmission electron microscopy(TEM)were used to evaluate changes in cell apoptosis and autophagy,respectively.The protein expression of LC3-II,p62,Bcl-2 and Bax in HT-22 cells were measured by Western bolt analysis.?Results? The CCK-8 detection found that HT-22 cell activity was significantly decreased by ox-LDL in a dose-dependent manner.The high dose concentration of ox-LDL(100 ?g/ml)could increase the apoptosis rate,upregulated the expression level of Bax and downregulated the expression level of Bcl-2 and induced cell shrinkage,mitochondria swelling,and chromatin condensation.Our results also revealed that co-treatment with CQ slightly increased LC3-II and p62 levels in ox-LDL-treated HT-22 cells,while co-treatment with RAP could elevate LC3-II and lower p62 levels in ox-LDL-treated HT-22 cells.These results indicating that ox-LDL treatment caused a defective autophagy flux in HT-22 cells.RAP treatment could reverse the adverse influences on HT-22 cells induced by ox-LDL,including the decline of cell viability,an increase in the cell apoptosis rate,upregulation of Bax,and downregulation of Bcl-2 expression level.In contrast,the ox-LDL induced the damage effects,as mentioned above,were aggravated by co-treatment with CQ.These results implied that activation of autophagy could protect HT-22 cells from cell apoptosis induced by ox-LDL.