CAJ | 학술논문

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500 μmol/L H2O2在37℃条件下进行)和依达拉奉组(提前30 min加入终质量浓度为300μmol/L的依达拉奉后加入终质量浓度为500 μmol/L H-2O2,在37℃条件下进行).免疫荧光显微镜观察神经元形态、Western blot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 H2 O2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.
목적 관찰의체랍봉통과c-jun안기단격매(JNK)통로화선립체신호도경대신경원조망적보호작용.방법 채용신생1dSD유대서체외배양적대뇌피층신경원피수궤분위공백대조조、양화응격조(가입종질량농도위500 μmol/L H2O2재37℃조건하진행)화의체랍봉조(제전30 min가입종질량농도위300μmol/L적의체랍봉후가입종질량농도위500 μmol/L H-2O2,재37℃조건하진행).면역형광현미경관찰신경원형태、Western blot법검측JNK、P-JNK상관단백표체、검측세포색소C(Cyt-C)화선립체형태적변화.결과 의체랍봉조중0.5h화2.0h량개양본적P-JNK/JNK비치분별위0.057화0.172,상대우양화응격조명현강저,차이유통계학의의(P＜0.05).재Cyt-C적검측중,의체랍봉조중0.5h화2.0h량개양본적Cyt-C분포교동시간점적양화응격조집중,차전경중야가견의체랍봉조(0.5h、2.0h)중선립체손상비동시간점적양화응격조경.결론 H2 O2유도적양화응격반응가능통과JNK통로화선립체신호도경유발신경원세포적조망,재조망조기사용의체랍봉가능통과JNK통로화선립체신호도경억제신경원조망.
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 of500 μmol/L H2O2 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 500 μ mol/L H2 O2 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 cytochrome (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 signaling pathways.The use of edaravone in the early stage of apoptosis may inhibit neuronal mitochondrial apoptosis through the JNK pathway and mitochondrial signaling pathway.