摘要
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.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2012年第5期937-939,共3页
Chinese Journal of Experimental Surgery
基金
国家自然科学基金资助项目(30973030)
江苏省卫生厅开放课题
关键词
神经元
脱噬作用
线粒体
活性氧
依达拉奉
Neuron
Apoptosis
Mitochondria
Reactive oxygen species
Edaravone