阿魏酸钠对SNP诱导的凋亡海马神经元NF-κB、par-4基因表达的影响

Effect of sodium ferulate on NF-κB P65 and Par-4 expression of apoptotic hippocampal neurons induced by SNP

目的探讨阿魏酸钠(sodium ferulate,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NF-κBP65、par-4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/L的SF预处理后,用50μmol/L的SNP处理24h,采用MTT法检测细胞存活率,Hochest33258荧光染色检测凋亡,Westernblot及RT-PCR检测NF-κBP65、par-4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;降低NF-κBP65、par-4mRNA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,其机制可能与降低促凋亡基因NF-κBP65、par-4表达有关。

Objective To investigate the protective effects of sodium ferulate （SF） on apoptosis of cultured hippocampal neurons in- duced by nitric oxide-donor, sodium nitroprusside （SNP） and the effect of SF on expression of NF-KB P65 and par-4. Methods The primary cultured hippocampal neurons were exposed to 50 μmol SNP 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 Hochest 33258 was used to analyze apoptosis. The expressions of NF-KB P65, par-4 mRNA and protein were tested by RT-PCR and Western blot. Results Pretreatment with SF（ 10-160 μmol/ml） for 6 h increased the survival rate of neurons. SF prevented the neuronal nuclei from shrinkage, condensation and cleavage induced by SNP. SF also decreased the expressions of mRNA and protein of NF-KB P65 and par-4. Conclusions SF can prevent the cultured hippocampal neurons against SNP neurotoxicity. The mechanism of protection is likely related to decreasing the level of NF-KB P65 and par-4.