阿魏酸钠通过JNK信号传导通路对抗Aβ_(1-42)引起的海马神经元凋亡

Sodium ferulate inhibits Aβ_(1-42)-induced hippocampal neuronal apoptosis through inhibition of c-Jun N-terminal kinases signaling pathways

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、200μmol.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(5μmol.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、200μmol.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(100μmol.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 hipp- ocampal 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 ap- optosis 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 ob- serve the level of of Bcl-2, Bax, active Caspase-3 and phospho-JNK in cultured hippocampal neurons. Re- suits 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 ap- optosis by inhibiting JNK signaling cascade.