【摘要】 目的观察依达拉奉后处理(EPC)对氧糖剥夺损伤SK-N-SH细胞的抗凋亡作用及对Bcl-2与Bax表达的影响,探讨其可能的机制。方法细胞分为正常对照组、氧糖剥夺组(OGD组)、依达拉奉后处理组(EPC组)。正常对照组DMEM高糖培养液中正常培养;OGD组的SK-N-SH细胞换用无糖DMEM培养液并置入自制缺氧罐处理4 h,再换成DMEM高糖培养液继续培养20 h;EPC组的SK-N-SH细胞先氧糖剥夺4 h,然后用含终浓度1μmol/L依达拉奉的DMEM高糖培养液作为后处理4 h后,再换成DMEM高糖培养基继续培养16 h。四甲基偶氮唑蓝(MTT)法检测细胞存活率,Hoechst33258染色检测细胞凋亡,Western blot检测Bcl-2与Bax蛋白的表达。结果 OGD组存活率比正常组明显下降(P<0.01),EPC组细胞存活率比OGD组显著增加(P<0.05);OGD组细胞凋亡率较正常组显著增高(P<0.01),而EPC组细胞凋亡率显著低于OGD组(P<0.05);与OGD组比较,EPC组Bcl-2表达显著增高(P<0.05),而Bax的表达明显降低(P<0.05)。结论依达拉奉后处理对氧糖剥夺SK-N-SH细胞有抗凋亡作用,可能与其使Bcl-2过表达和抑制Bax表达有关。更多还原

【Abstract】 Objective In the present study,we investigated the effect of edravone postconditioning( EPC) on SK-NSH cells anti-apoptosis in oxygen-glucose deprivation( OGD) model and regulation of Bcl-2 / Bax expression. Methods 4hrs OGD was induced by OGD experiments. To initiate OGD,culture medium was replaced with deoxygenated,glucose-free DMEM medium. After 4 hrs’ challenge,the SK-N-SH cells were then allowed to recover for 20 hrs in the glucose-high DMEM medium. For the EPC group,the cells were incubated by culture medium containing final concentrations of 1 μmol / L edaravone for 4hrs after 4 hrs’ OGD experiments. Cells were then allowed to recover for 16 hrs in the normal condition. Viability and apoptotic population of SK-N-SH cells were quantified,respectively,by MTT( 2-( 4,5-dimethylthia-zol-2-yl)-2,5-diphenyltetrazolium bromide) detecting and Hoechst 33258 staining. In addition,Bcl-2 and Bax expression were examined by western blot. Results Compared to Normol,cell viability of OGD group decreased by approximately 43%( P < 0. 01),but that of EPC group was significantly increased( P = 0. 023),compared to OGD group. The rate of apoptosis of EPC group was notably lower than that of OGD group( P = 0. 011). Bcl-2 expression of EPC group was predominately increased( P = 0. 012),whereas Bax expression of EPC group observably decreased( P = 0. 016),contrasted with OGD group. Conclusion The effect of EPC on SK-N-SH cells after OGD may be achieved through the inhibition of apoptosis,which possible mechanism may be related to promote Bcl-2 expression and inhibit Bax expression.更多还原