原花青素对Aβ_(25-35)诱导PC12细胞周期紊乱的保护作用

Study on protective effects of Proanthocyanidins on abnormal cell cycle in PC12 cells induced by β-amyloid peptide 25-35

目的观察原花青素(PAC)对β-淀粉样肽(25-35)(Aβ25-35)诱导体外血清饥饿培养的PC12细胞周期紊乱与凋亡的保护作用,并探讨其可能的作用机制。方法采用血清饥饿培养使PC12细胞同步于G0期,PAC预处理后加入Aβ25-35,通过流式细胞仪分析细胞周期与凋亡,RT-PCR和Western印迹从mRNA及蛋白水平检测p21、Cyclin D1、pRb和E2F1基因表达的变化。结果与Aβ25-35诱导组比较,PAC保护组S期细胞百分率明显减少,凋亡率明显降低(P<0.05);p21 mRNA和蛋白表达增加(P<0.05),Cyclin D1、E2F1 mRNA和Cyclin D1、pRb蛋白表达降低(P<0.05)。结论PAC对Aβ25-35诱导的PC12细胞周期紊乱与凋亡起保护作用,其机制可能与上调p21表达,下调Cyclin D1、pRb、E2F1表达有关。

Objective To observe the protective effects of Proanthocyanidins （PAC） on abnormal cell cycle in PC12 cells induced by β-amyloid peptide 25-35 （Aβ25-35）and study its possible mechanisms. Methods PC12 cells were synchronized by 24 h serum deprivation, pretreated with PAC for 1 h, and then Aβ25 -35 for 0 -24 h. The changes of cell cycle distribution and apoptosis were analyzed by flow eytometry. Protein and mRNA expressions of p21, Cyelin D1 ,pRb and E2F1 were detected by RT-PCR and Western-blot respectively. Results Compared with those in Aβ25-35 group, the percent of S phase cells and apoptotie rate were decreased remarkably （ P 〈 0. 05 ） in PAC group, the mRNA and protein expressions of p21 gene were increased ,and the expressions of Cyelin D1 ,pRb and E2F1 gene were decreased （ P 〈 0. 05 ）. Conclusions PAC has protective effects of redistribution of cell cycle and apoptosis in serum-deprived PC12 cells induced by A132s_35, which is related with up-regulating p21 expression, and down-regulating Cyelin D1 ,pRb and E2F1 gene expressions.