阿魏酸钠对SNP诱导的凋亡海马神经元bcl-2、bax基因表达的影响

Effect of sodium ferulate on bcl-2 and bax gene expression of apoptotic hippocampal neurons induced by sodium ferulate

目的:探讨阿魏酸钠(SF)对一氧化氮(NO)供体硝普钠(SNP)引起的大鼠海马神经元凋亡及bcl-2、bax基因表达的影响。方法:采用SD大鼠海马神经元原代培养,经终浓度分别为10、20、40、80、120、160μmol/LSF预处理后,用50μmol/L SNP处理24h,采用MTT法检测细胞存活率,Hoechst33258荧光染色及DNA琼脂糖凝胶电泳分析等方法检测凋亡,Western blotting及RT-PCR检测bcl-2、bax基因表达。结果:不同剂量SF(10-160μmol/L)预处理6h可显著提高神经元的存活率,减少SNP引起的核固缩、凝聚和碎裂现象;DNA凝胶电泳图谱未见典型的"梯状"改变;增加bcl-2mRNA及蛋白的表达,降低baxmRNA及蛋白的表达。结论:SF抑制NO供体SNP诱导的海马神经元凋亡,其机制可能与其增加Bcl-2蛋白表达,降低Bax蛋白表达,增高Bcl-2/Bax的比值有关。

AIM： To investigate the protective effects of sodium ferulate （SF） on apoptosis in cultured hippocampal neurons induced by sodium nitropmsside （SNP） , and the effect of SF on expression of bcl- 2 and bax. METHODS ： The primary cultured hippocampal neurons were exposed to 50μmol SNP, a nitric oxide - donor, for 24 h after pretreatment with different concentrations of SF （ 10 - 160μmol/mL） for 6 h. Then neuronal viability was tested by MTT assay. Fluorescent staining with Hoechst 33258 and agarose gel electrophoresis was used to analyze apoptosis. The expressions of bcl -2, bax mRNA and protein were tested by RT- PCR and Western blotting. RESULTS： Pretreatment with SF （ 10 - 160 μmol/L） for 6 h increased the survival rate of neurons. SF prevented the neuronal nuclei from shrinkage, condensation and cleavage and blocked neuronal nuclear DNA fragmentation induced by SNP. SF also increased the expressions of bcl - 2 mRNA and Bcl - 2 protein and decreased the expressions of bax mRNA and Bax protein. CONCLUSION ： SF prevents the cultured hippocampal neurons against SNP neurotoxicity. The mechanism of protection is related to the increase in Bc1-2 level and the decrease in Bax level. As a result, the ratio of Bcl -2/Bax is changed.