硝普钠对海马神经元cpp32基因表达的影响

Effects of sodium nitroprusside on the expression of hippocampal neuron gene cpp 32

为观察NO供体硝普钠对体外培养的海马神经元cpp32基因表达的影响 ,采用终浓度分别为 0、2 5、5 0、10 0、2 0 0、4 0 0 μmol L的硝普钠处理海马神经元 2 4h ,用RT PCR检测mRNA表达变化 ,Westernblot检测蛋白表达的变化 ;再用终浓度分别为 0、2 5、5 0、10 0、2 0 0 μmol L的硝普钠处理海马神经元 12h ,用CPP32活性检测试剂盒检测CPP32酶活性。结果表明 ,随着硝普钠剂量的增加 ,cpp32mRNA表达无改变 ;但CPP32酶原被裂解活化 ,从 5 0 μmol L硝普钠起 ,酶活性显著增加 ,为对照组的 3 0 2倍 ,10 0 μmol L达最大值 ,为对照组的 3 4 7倍 ,这说明硝普钠不增加cpp32mRNA的表达 ,但可诱导CPP32酶原的裂解 ,使CPP32活化。

To determine the effects of nitric oxide donor, sodium nitroprusside(SNP) on the expression of gene cpp 32 in the cultured Hippocampl neurons in vitro, the cultured Hippocampl neuron were treated with different concentrations of SNP(0～400μmol/L) for 24h , and the expression of mRNA and protein of these neurons were analyzed with RT PCR and Western blot respectively. The cultured Hippcampl neuron was treated with different concentrations of SNP(0～200μmol/L) for 12h and the activity of cpp 32 was tested. The results showed that the cpp 32 mRNA level was unchanged following the increasing dose of SNP,but the pro Cpp32 was activated and the activity of cpp 32 increased significantly by 50μmol/L SNP which was 3.02 folds of that in control group and reached to the max value at 100μmol/L which was 3.47 folds of that in control group. The results indicated that SNP failed to increase Cpp32 mRNA expression but increased degradation of pro Cpp32 and activated pro Cpp32.