阿魏酸钠通过JNK信号传导通路对抗Aβ1-42引起的海马神经元凋亡

摘要

目的 研究阿魏酸钠(SF)对Aβ1-42所致培养海马神经元凋亡的抑制作用及机制.方法 原代培养海马神经元,SF(50、100、200 μmol·L-1)预处理6 h后,加入50 nmol·L-1的Aβ1-42作用72 h,JNK阻断剂SP600125(5 μmol·L-1)在加入SF前30 min加入,Hoechst33258核染色观察细胞凋亡变化,ELISA法检测细胞色素C释放量,Western蛋白印迹法检测神经元Bcl-2,Bax,Caspase-3及JNK 蛋白表达.结果 与对照组相比,Aβ1-42组海马神经元细胞的凋亡率及释放的细胞色素C含量明显增加(P<0.01),Bax与bcl-2蛋白表达比值,磷酸化的Caspase-3及磷酸化JNK蛋白表达明显增加 (P<0.01).应用SF(50、100、200 μmol·L-1)预处理6 h可明显对抗Aβ1-42引起的海马神经元细胞凋亡、细胞色素C释放量,SF(100 μmol·L-1)能抑制蛋白表达的改变(P<0.01),JNK阻断剂也能抑制Aβ1-42引起的这些改变.结论 SF通过抑制JNK信号传导通路对抗Aβ1-42引起的海马神经元损伤.%Aim To investigate the protective effect of sodium ferulate ( SF ) on apoptosis of cultured hippocampal neurons induced by Aβ1-42. Method The primary cultured hippocampal neurons were exposed to Aβ1-42 50 nmol · L-1 for 72 h after pretreatment with various concentrations of SF( 50, 100, 200 μmol · L-1 )for 6 h. SP600125 was added respectively to the cells 30 min prior to SF treatment. Then neuronal apoptosis was quantified by scoring the percentage of cells with apoptotic nuclear morphology after Hoechst 33258 staining. Cytosolic cytochrome C was measured by ELISA technology. Western blot was peformed to observe the level of of Bcl-2, Bax, active Caspase-3 and phospho-JNK in cultured hippocampal neurons. Results cultured hippocampal neurons incubated with Aβ1-42 significantly increased apoptotic rates and mito-chondrial release of cytochrome C, and also caused an increase in Bax, active Caspase-3 and phospho-JNK levels, and a decrease in Bcl-2 levels. These Aβ1-42-induced changes could be reversed by pretreatment with SF or SP600125. Conclusion SF can protect cultured hippocampal neurons from Aβ1-42-induced apoptosis by inhibiting JNK signaling cascade.