二氮嗪对氧糖剥夺后PC12细胞凋亡及对Bcl-2蛋白的影响

目的探讨二氮嗪对氧糖剥夺(OGD)PC12细胞凋亡的保护作用及其机制。方法体外培养PC12细胞株,以OGD、二氮嗪、5-羟葵酸(5-HD)处理,分为A组(正常对照组),B组(氧糖剥夺组),C组(氧糖剥夺+二氮嗪组),D组(氧糖剥夺+二氮嗪组+5-HD组),采用Annexin V-FITC/PI双染流式细胞分析仪检测凋亡率,应用免疫荧光染色和Western blot检测Bcl-2蛋白表达水平,观察二氮嗪对氧糖剥夺PC12细胞凋亡的保护作用。结果氧糖剥夺后B,C,D组PC12细胞凋亡细胞数增加,C组与B、D组比较差异均有显著性(P<0.01)。B,C,D组Bcl-2蛋白表达增加,C组达到高峰。C组与A、B、D组比较差异均有显著性(P<0.01)。结论二氮嗪能抑制氧糖剥夺PC12细胞凋亡,这一作用机制可能是增加Bcl-2表达来实现其保护作用的。

Effects of diazoxide on expression of Bcl-2 following oxygen-glucose deprivation in PC12 cell

Aim To study the effect of diazoxide on the apoptosis of PC12 cells induced by oxygen-glucose deprivation(OGD)and expression of Bcl-2 protein.Methods Cultured PC12 cells,treated with OGD,diazoxide and 5-HD,were divided into A(control group),B(OGD group),C(OGD+diazoxide group)and D(OGD+diazoxide+5-HD group).Neuronal apoptosis was detected by Annexin V-FITC/PI double-dyed flow cytometry,and the expression of Bcl-2 protein was detected by immunofluorescence and Western blot.Results The number of apoptotic PC12 cells increased after OGD in B,C,D group.C group and B,D groups were significantly different(P<0.01).The expression of Bcl-2 protein in B,C and D group was higher than that of A group,and that in C group was significant higher than that of A,B and D group(P<0.01).Conclusion Diazoxide has a protective effect on the apoptosis of PC12 cells induced by OGD,which might be correlated with the up-regulated expression of Bcl-2 protein.