Effect of sodium ferulate on bcl-2 and bax gene expression of apoptotic hippocampal neurons induced by sodium ferulate
Daoming Li
Abstract
Daoming Li
Abstract
AIM:To investigate the protective effects of sodium ferulate(SF)on apoptosis in cultured hippocampal neurons induced by sodium nitroprusside(SNP),and the effect of SF on expression of bcl-2 and bax.METHODS:The primary cultured hippocampal neurons were exposed to 50 μmol SNP,a nitric oxide-donor,for 24 h after pretreatment with different concentrations of SF(10-160 μmol/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 Bcl-2 level and the decrease in Bax level.As a result,the ratio of Bcl-2/Bax is changed.
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AIM:To investigate the protective effects of sodium ferulate(SF)on apoptosis in cultured hippocampal neurons induced by sodium nitroprusside(SNP),and the effect of SF on expression of bcl-2 and bax.METHODS:The primary cultured hippocampal neurons were exposed to 50 μmol SNP,a nitric oxide-donor,for 24 h after pretreatment with different concentrations of SF(10-160 μmol/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 Bcl-2 level and the decrease in Bax level.As a result,the ratio of Bcl-2/Bax is changed.
Key concepts: Apoptosis, Sodium nitroprusside, Molecular biology, Chemistry, Agarose gel electrophoresis, DNA fragmentation, Neurotoxicity, Hippocampal formation