依达拉奉通过c—jun氨基端激酶通路和线粒体信号途径对H2O2诱导神经元凋亡的保护作用

Protective effect of edaravone on H2O2,induced neuron apoptosis via c-Jun N-terminal kinase patheway and mitochondrial signaling patheway

目的观察依达拉奉通过c-jun氨基端激酶（JNK）通路和线粒体信号途径对神经元凋亡的保护作用。方法采用新生1dSD乳大鼠体外培养的大脑皮层神经元被随机分为空白对照组、氧化应激组（加入终质量浓度为500tzmol／LH2O3在37℃条件下进行）和依达拉奉组（提前30min加入终质量浓度为300μmol／L的依达拉奉后加入终质量浓度为500tunol／LH2O2，在37℃条件下进行）。免疫荧光显微镜观察神经元形态、Westernblot法检测JNK、P—JNK相关蛋白表达、检测细胞色素c（Cyt—C）和线粒体形态的变化。结果依达拉奉组中0．5h和2．0h两个样本的P-JNK／JNK比值分别为0．057和0．172，相对于氧化应激组明显降低，差异有统计学意义（P〈0．05）。在Cyt—C的检测中，依达拉奉组中0．5h和2．0h两个样本的cyt—C分布较同时间点的氧化应激组集中，且电镜中也可见依达拉奉组（0．5h．2．0h）中线粒体损伤比同时间点的氧化应激组轻。结论H2O2诱导的氧化应激反应可能通过JNK通路和线粒体信号途径诱发神经元细胞的凋亡，在凋亡早期使用依达拉奉可能通过JNK通路和线粒体信号途径抑制神经元凋亡。

Objective To investigate the protective effects of edaravone on neuronal apoptosis through the c-Jun N-terminal kinase （JNK） pathway and the mitochondrial signaling pathway. Methods Newborn 1 day SD neonatal rat cultured cortical neurons were randomly assigned to 3 groups： blank control group, the oxidative stress group （final concentration of 500 μmol/L H202 at 37 ℃） and edaravone group （ added to a final concentration of 300 μmol/L of edaravone 30 min earlier, then adding a final concentration of 500μmol/L H2O2 at 37 ℃ ）. The neuronal morphology was observed under an immunofluorescence microscope. The protein expression of JNK and P-JNK was etected by using Western blotting. Results The P-JNK/JNK ratio of two samples （0. 5 h and 2. 0 h） in edaravone group was 0. 057 and 0. 172 respectively, wich was significantly reduced as compared with the oxidative stress group （ P 〈 0. 05 ）. The cyto-chrome （Cyt-C） distribution of two samples （0. 5 h and 2.0 h） in edaravone group was more concentrated than in the oxidative stress group. The mitochondrial oxidative damage of two samples （0. 5 h and 2. 0 h） in oxidative stress group was more serious than in edaravone group at the same time. Conclusion H2O2-induced oxidative stress may induce neuronal apoptosis through the JNK pathway and mitochondrial signa-ling pathways. The use of edaravone in the early stage of apoptosis may inhibit neuronal mitochondrial ap-optosis through the JNK pathway and mitochondrial signaling pathway.