藻酸双酯钠对蜂毒肽诱导的皮质神经元凋亡的保护作用

Polysaccharide sulphate blocks apoptosis induced by mastoparan in cortical neurons

目的 研究藻酸双酯钠（ＰＳＳ） 对蜂毒肽（ｍａｓｔｏｐａｒａｎ）诱导的皮质神经元凋亡是否具有保护作用。方法 体外培养大鼠皮质神经元，定性定量检测细胞凋亡：①用ＦＤＡ（ｆｌｕｏｒｅｓｃｅｉｎ ｄｉａｃｅｔａｔｅ） 染色检测细胞存活率，用Ｈｏｅｃｈｓｔ ３３２５８ 染色，分析细胞核的形态变化；②琼脂糖凝胶电泳分析ＤＮＡ 断裂；③流式细胞仪对细胞凋亡峰进行定量分析。结果 ＰＳＳ使皮质神经元存活率增加，使ｍａｓｔｏｐａｒａｎ 引起的细胞核固缩、凝聚和断裂现象消失；ｍａｓｔｏｐａｒａｎ 使神经元的ＤＮＡ电泳图谱出现明显的“梯子状”，而ＰＳＳ使此现象消失；ｍａｓｔｏｐａｒａｎ使神经元的流式细胞仪分析图上出现明显的凋亡峰，ＰＳＳ可使此凋亡峰消失。

AIM To study whether polysaccharide sulphate (PSS) can block cortical neuronal apoptosis induced by mastoparan. METHODS Three methods were used to investigate the protective effects of PSS on cultured rat cortical neurons damaged by mastonaran:①The neurons were stained with fluorescein diacetate to observe their survival rate or stained with Hoechst 33258 to analyze their nuclear morphology. ②Internucleosomal DNA fragmentation of cortical neurons was analyzed with agarose gel electrophoresis.③Flow cytometry was used to analyze neuronal apoptotic peak quantitatively. RESULTS PSS increased the survival rate of cortical neurons damaged by mastoparan in a dose dependent manner and prevented the nuclei from shrinkage, condensation and cleavage induced by mastoparan. PSS prevented neuronal nuclear DNA fragmentation induced by mastoparan. PSS blocked the neuronal apoptotic peak induced by mastoparan. CONCLUSION PSS can block cortical neuronal apoptosis induced by mastoparan.