摘要
Background： Edaravone had been validated to effectively protect against ischemic injuries. In this study, we investigated the protective effect of edaravone by observing the effects on anti-apoptosis, regulation of Bcl-2/Bax protein expression and recovering from damage to mitochondria after OGD （oxygen-glucose deprivation）-reperfusion. Methods： Viability of PC 12 cells which were injured at different time of OGD injury, was quantified by measuring MTT （2-（4,5-dimethylthia-zol-2-yl）-2,5-diphenyltetrazolium bromide） staining. In addition, PC 12 cells＇ viability was also quantified after their preincubation in different concentration of edaravone for 30 min followed by （OGD）. Furthermore, apoptotic population of PC 12 cells that reinsulted from OGD-reperfusion with or without preincubation with edaravone was determined by flow cytometer analysis, electron microscope and Hoechst/Pl staining. Finally, change of Bcl-2/Bax protein expression was detected by Western blot. Results： （1） The viability of PC12 cells decreased with time （1-12 h） after OGD. We regarded the model of OGD 2 h, then replacing DMEM （Dulbecco＇s Modified Eagle＇s Medium） for another 24 h as an OGD-reperfusion in this research. Furthermore, most PC 12 cells were in the state of apoptosis after OGD-reperfusion. （2） The viability of PC 12 cells preincubated with edaravone at high concentrations （1, 0.1, 0.01 μmol/L） increased significantly with edaravone protecting PC 12 cells from apoptosis after OGD-reperfusion injury. （3） Furthermore, edaravone attenuates the damage of OGD-reperfusion on mitochondria and regulated Bcl-2/Bax protein imbalance expression after OGD-reperfusion. Conclusion： Neuroprotective effects of edaravone on ischemic or other brain injuries may be partly mediated through inhibition of Bcl-2/Bax apoptotic pathways by recovering from the damage of mitochondria.
背景：Edaravone 被验证了有效地免于是化学家损害。在这研究，我们由在 anti-apoptosis 上观察效果调查了 edaravone 的保护的效果， Bcl-2/Bax 的规定蛋白质表示；在 OGD (氧葡萄糖剥夺) 以后从损坏恢复到线粒体 -reperfusion。方法：在 OGD 损害的不同时间被伤害的 PC12 房间的生存能力，被测量染色的 MTT (2-(4,5-dimethylthiazol-2-yl )-2,5-diphenyltetrazolium 溴化物) 确定。另外，在他们在为 30 min 的 edaravone 的不同集中的 preincubation 列在后面由以后， PC12 房间的生存能力也被确定(OGD ) 。而且，与 edaravone 从 OGD-reperfusion 重新侮辱与或没有 preincubation 的 PC12 房间的 apoptotic 人口被流动 cytometer 分析决定，电子显微镜；染色的 Hoechst/PI。最后， Bcl-2/Bax 蛋白质表示的变化被西方的污点检测。结果：(1 ) PC12 房间的生存能力与时间减少了(1 ～ 1 2 h ) 在 OGD 以后。我们考虑了 OGD 2 h 的模型，然后在这研究作为 OGD-reperfusion 为另一 24 h 代替 DMEM (Dulbecco 的修改的鹰的媒介) 。而且，大多数 PC12 房间处于在 OGD-reperfusion 以后的 apoptosis 的状态。(2 ) 有在高集中的 edaravone 的 PC12 房间 preincubated 的生存能力(1， 0.1， 0.01 μ m ol/L ) 与在 OGD-reperfusion 损害以后保护 PC12 房间免受 apoptosis 的伤害的 edaravone 显著地增加了。(3 ) 而且， edaravone 在线粒体上稀释 OGD-reperfusion 的损坏；在 OGD-reperfusion 以后的调整 Bcl-2/Bax 蛋白质不平衡表示。结论：edaravone 在上的 Neuroprotective 效果是化学家或另外的大脑损害可以被从线粒体的损坏恢复部分通过 Bcl-2/Bax apoptotic 小径的抑制调停。
基金
Project (No. 2005C30059) supported by the Science and TechnologyProgram of Zhejiang Province, China
