SNP对体外培养的海马神经元bcl-2和bax基因表达的影响

SNP INDUCES THE APOPTOSIS OF CULTURED RAT HIPPOCAMPAL NEURONS BY CHANGING THE EXPRESSION OF BCL-2 AND BAX GENE

目的 :探讨NO供体硝普钠 (SodiumNitroprusside,SNP)诱导体外培养的海马神经元凋亡与Bcl- 2、Bax表达变化的关系。方法 :终浓度分别为 0、2 5、5 0、10 0、2 0 0、4 0 0 μmol/L的SNP处理海马神经元 2 4h ,或用终浓度为 5 0 μmol/L的SNP分别处理海马神经元 0、6、12、18、2 4h ,用RT -PCR检测Bcl- 2、baxmRNA表达变化 ,Westernblot检测Bcl- 2、Bax蛋白表达的变化。结果 :随着SNP剂量的增加 ,bcl- 2mRNA、Bcl- 2蛋白表达逐渐减少 ,而baxmRNA、Bax蛋白表达逐渐增加 ;随着时间的处长 ,bcl- 2mRNA、Bcl- 2蛋白表达逐渐减少 ,而baxmRNA、Bax蛋白表达去逐渐增加。结论 :SNP可时间、剂量依赖性地降低bcl- 2mRNA、Bcl- 2蛋白的表达 ,时间、剂量依赖性地增加baxmRNA、Bax蛋白的表达。SNP诱导海马神经元凋亡的机制 ,可能是与降低Bcl- 2 /Bax的比值有关。

Objective:This study was designed to discuss the relationship between apoptosis of cultured rat hippocampal neurons induced by nitric oxide-donor, sodium nitroprusside (SNP) and the changes in Bcl-2 and Bax expression. Methods:The cultured hippocampal neurons were treated with different concentrations of SNP (0～400μmol/L) for 24h, or with SNP 50μmol/L for the indicated time,the expression changes of bcl-2 and bax mRNA were examined by PT-PCR and the expression changes of Bcl-2 and Bax protein were analyzed by Western blot. Results:As the increasing of SNP concentration , the expressions of bcl-2 mRNA and Bcl-2 protein decreased. However, the expressions of bax mRNA and Bax protein increased. As the prolonging of the time, the expressions of bcl-2 mRNA and Bcl-2 protein decreased. Whereas the expressions of bax mRNA and Bax protein increased. Conclusions:SNP decreased the expression of bcl-2 mRNA and Bcl-2 protein in a time-and dose-independent manner. Whereas it increased the expressions of bax mRNA and Bax protein in a time- and dose- independent manner. The mechanism of cultured rat hippocampal neuronal apoptosis induced by SNP is likely related to the decreasing of the ratio of Bcl-2/Bax.