原花青素对PC12细胞氧化应激损伤保护作用的细胞周期调控机制

Cell cycle regulation involved in protective effect of proanthocyanidins on PC12 cells damage from hydrogen peroxide

目的观察原花青素对过氧化氢诱导体外培养大鼠嗜铬细胞瘤PC12细胞氧化损伤及细胞周期阻滞的保护作用,并探讨细胞周期调控基因c-Abl和plk1在其中的作用。方法采用MTT法观察不同浓度原花青素对100μmol/L过氧化氢诱导6 h的PC12细胞增殖率的影响。采用流式细胞仪检测原花青素对诱导的PC12细胞周期分布的影响,Western blot检测不同浓度原花青素对诱导的PC12细胞c-Abl和plk1基因蛋白表达水平的影响。结果与对照组比较,过氧化氢显著降低PC12细胞增殖率(P<0.01)、显著增高PC12细胞S期分布比例(P<0.01)、显著诱导c-Abl和plk1基因蛋白水平表达增强(P<0.01)。与过氧化氢诱导组比较,原花青素剂量依赖性提高PC12细胞的增殖率(P<0.01)、缓解过氧化氢诱导的细胞周期S期阻滞(P<0.01)、降低c-Abl和plk1基因蛋白水平的表达(P<0.01)。结论原花青素对过氧化氢诱导的PC12细胞氧化损伤具有保护作用,其机制可能是通过缓解过氧化氢诱导的细胞周期S期阻滞、降低细胞周期调控相关基因c-Abl和plk1的蛋白表达水平来实现的。

Objective To determine the protective effect of proanthocyanidins on oxidative damage and cell cycle arresting in PC12 cells induced by hydrogen peroxide, and to investigate the role of cell cycle regulatory genes cellular-Abelson gene （c-Abl） and polo-like kinase-1 （plk1） in the protection. Methods PC12 cells were incubated with different concentrations of proanthocyanidins and induced by 100 μmol/L hydrogen peroxide for 6 h, and then the cell viability was measured by MTT assay, and the distribution of cell cycle was measured by flow cytometry, and the protein expression of c-Abl and plkl was detected by Western blotting. Results Compared with the PC12 cells in control group, hydrogen peroxide significantly decreased cell viability and increased S phase distribution in PC12 cells （P 〈0.01 ）, and the protein expression level of c-Abl and： plkl gene in PC12 cells was also increased significantly by hydrogen peroxide （P 〈 0. 01 ）; Compared with the group of PC12 cells induced by hydrogen peroxide, proanthocyanidins significantly increased cell viability of PC12 cells in the range of experimental doses （ P 〈 0.01 ）. Proanthocyanidins relieved the S phase arrest induced by hydrogen peroxide （P 〈0.01 ）, and the protein levels of c-Abl and plkl in PC12 cells was also decreased by proanthocyanidins in a dosedependent manner （P 〈 0. 01 ）. Conclusion Proanthocyanidins can protect PC12 cells from the cytotoxic stresses induced by hydrogen peroxide, and the protective mechanism may be mediated by relieving the S phase arrest and decreasing the protein expressional level of c-Abl and plkl gene.