酸性肽对硝普钠诱导的大鼠海马神经细胞凋亡中Bcl-2和Bax表达的影响

Effects of acidic peptide on the expression of Bcl-2 and Bax in apoptotic hippocampal neurons induced by SNP

目的观察不同剂量酸性肽(AP)对NO供体硝普钠(SNP)诱导的大鼠海马神经细胞凋亡中bcl-2和bax基因表达的影响,探讨酸性肽抑制神经细胞凋亡的机制。方法分离培养新生大鼠海马神经细胞,用SNP诱导海马神经细胞凋亡,采用吖啶橙荧光染色、DNA琼脂糖凝胶电泳、Westernblot研究加入SNP培养的及SNP和AP共培养的海马神经细胞的形态和分子生物学改变及Bcl-2和Bax蛋白表达的变化。结果AP能对抗SNP诱导的凋亡,改善凋亡细胞的形态学改变;减少SNP诱导的海马神经细胞核固缩、核碎裂现象,抑制DNA凋亡梯带的出现,上调Bcl-2的表达而下调Bax的表达。结论AP能抑制SNP诱导的海马神经细胞凋亡,其机制可能通过调控Bcl-2和Bax的表达来实现。

[Objective] To discuss the mechanism of AP protective function to apoptotic neurons, this study was designed to observe the effect of AP on the expression of Bcl-2 and Bax in apoptotic hippocampal neurons triggered by sodium nitroprusside （SNP）. [Methods] The new born hippocampal neurons were cultured in serum free medium with 50 μmol/LSNP for establishing neuronal apoptosis model. Adding different concentrations of AP to cultured cell for 24h. Morphological and biochemical change were observed by fluorescent staining and DNA agarose gel electrophoresis. Western blot was used to analyze the expression changes of Bcl-2 and Bax. [Results] AP improved the morphologic changes and prevented the neuronal neclei from shrinkage, condensation and cleavage triggered by SNP. It also blocked neuronal nuclear DNA fragmentation elicited by SNP. AP up-regulated the expression of Bcl-2 and down-regulated the expression of Bax in a concentration dependent fashion. [Conclusion] The mechanism of AP inhibiting neuronal apoptosis induced by SNP is likely involved in regulating the expression of Bcl-2 and Bax.