目的探讨原花青素(PC)对β-淀粉样肽(Aβ_25-35)诱导凋亡PC12细胞par-4和bcl-2基因mRNA和蛋白表达的影响。方法采用MTT比色法分析细胞存活率, Hoechst33258-PI荧光染色法检测细胞凋亡, RT-PCR方法检测PC12细胞par-4和bcl-2基因mRNA的表达, Westernblotting检测PC12细胞Par-4和Bcl-2蛋白表达。结果不同剂量(5、10、20、30mg/L)PC预处理PC12细胞1h可剂量依赖性对抗Aβ_25-35引起的细胞凋亡, 提高PC12细胞的存活率, 减少Aβ_25-35引起的PC12细胞核固缩、凝聚和碎裂, 降低par-4基因mRNA表达及蛋白表达, 增加bcl-2基因mRNA表达及蛋白表达。结论PC可剂量依赖性对抗Aβ_25-35对PC12细胞的毒性作用, 其机制可能与下调凋亡基因par-4和上调抗凋亡基因bcl-2表达有关。

Objective To study the effect of procyanidins(PC)on mRNA and protein expression of par-4 and bcl-2 genes in PCI2 cells induced by Aβ_25-35.Methods Cell survivaI rate was evaluated by MTT assay and apoptosis was analyzed by Hoechst 33258-PI fluorescence staining.The expressions of mRNA and protein for par-4 and bcl-2 were tested by RT-PCR and Western blotting.Results Pretreatment with different concentrations of PC(5、10、20,and 30 mg/L)for l h increased the survivaI rate of PCI2 celI in a dose-dependent manner.PC prevented the PCI2 cells nuclei from shrinkage,condensation,and cleavage induced by Aβ_25-35.PC decreased the expression of par-4 mRNA and protein, and increased the expression of bcl-2 mRNA and protein.Conclusion PC can protect PCI2 cells from apoptosis induced by Aβ_25-35 in a dose dependent manner.The mechanism of protection is likely related to decreasing the par-4 gene expression and increasing the bcl-2 gene expression.

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