吡那地尔对缺血缺氧PC12细胞凋亡及Bcl-2蛋白表达的影响

目的探讨吡那地尔对缺血缺氧PC12细胞凋亡及对Bcl-2蛋白表达的影响。方法取传代后3d PC12细胞，分为A（对照组），B（缺血缺氧组），C（KATP通道开放剂），D（KATP通道开放剂＋阻断剂组）。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。结果缺血缺氧后B，C，D组细胞凋亡率随时间点增加而增加，24h达高峰。B，C，D组与A组比较均有显著性差异（P〈0．01），C组和B，D组比较有统计学意义（P〈0．01），B，C，D组细胞Bcl-2蛋白表达随时间点增加而增加，12h达高峰。B，C，D组均显著高于对照组（P〈0．01或P〈0．05）。C组表达显著高于B和D组（P〈0．01）。B与D组各时间点细胞凋亡及Bcl—2蛋白表达均无显著性差异（P〉0．05）。结论KATP通道开放剂能抑制缺血缺氧PC12细胞凋亡，这一作用机制可能与增加Bcl—2蛋白表达有关。

EFFECTS OF PINACIDIL ON THE APOPTOSIS OF PC12 CELLS INDUCED BY ISCHEMIA AND HYPOXIA AND THE EXPRESSION OF BCL-2 PROTEINS

ObjectiveTo study the effects ofpinacidil on the apoptosis of PC12 cells induced by ischemia and hypoxia and on the expression of Bcl-2 proteins. Methods Cultured PC12 cells were divided into 4 groups： A（control）, B（ischemia and hypoxia）, C（K^ATP opener） and D（K^ATP opener and blocker treatment）. Neuronal apoptosis was detected by Annexin-v FITC/PI double-dyed flow cytometry, and the expression of Bcl- 2 proteins was detected by Immunofluorescent staining and Western blotting.Results The apoptotic ratio in groups B,C and D increased peaking at 24 hours. There were significant differences between groups B, C, D and A （P〈0.01） . and between groups C and B, D.（P〈0.01）. The expression of Bcl-2 protein in groups B,C and D is also increased with time, peaking at 12 hours It was significanly higher in group B, C and D than in group A （P〈0.05, or P〈0.01）, and significantly higher in group C than in groups B and D （P〈0.05, or P〈0.01） . There were no differences between groups B and D at any time points （P〉0.05） .Conclusion K＋ATP opener has a protective effect on the apoptosis of PC 12 ceils induced by ischemia and hypoxia ,which may be correlated with the up-regulated expression of Bcl- 2 protein.