芡实正丁醇部位分离组分对H2O2诱导的PC12细胞的神经保护作用及其机制研究

Neuro-Protective Effect and the Mechanism of Semen Euryales Active Fraction Against H2O2-Induced PC12 Cells

  • 摘要: 目的 研究芡实正丁醇部位的聚酰胺柱分离组分对H2O2损伤的PC12细胞的神经保护作用,探讨其对阿尔茨海默病潜在的预防治疗作用及其可能的机理。方法 体外培养PC12细胞株,加入浓度为200 μmol/L的H2O2损伤细胞8 h,建立氧化损伤模型,将芡实不同浓度的乙醇提取物分别作用于细胞后,采用MTT法检测各组分对细胞存活率的影响,用试剂盒测定细胞液中SOD活力、MDA含量,RT-PCR检测细胞内APP、BACE-1、GSK-3β的mRNA表达水平。结果 与模型组相比,芡实正丁醇部位50%、70%、95%乙醇的洗脱物均能显著改善H2O2损伤的PC12细胞的存活率(P<0.05),70%、95%乙醇的洗脱物均能显著增加细胞液中SOD酶的活力,减少MDA的释放(P<0.05),下调GSK-3β mRNA表达量(P<0.01)。结论 芡实正丁醇聚酰胺柱70%、95%的乙醇洗脱物对氧化应激损伤的PC12细胞具有显著的保护作用,其作用机制可能与增加SOD酶活力、减少MDA释放、抑制Aβ累积相关的GSK-3β基因表达有关。

     

    Abstract: OBJECTIVE To investigate the effect of n-butanol extract from Euryale ferox Salisb. against H2O2-induced oxidative injury of PC12 cells and explore the protective mechanism of this active components against Alzheimer's disease cell model. METHODS The n-butanol extracts of semen euryales were separated by the column chromatography of polyamide. PC12 cells were cultivated in vitro and treated with 200 μmol/L H2O2 for 8 h to establish the oxidative damage model. Cell viabilities were determined by MTT assay, and the activities of superoxide dismutase (SOD) and the content of malondialdehyde (MDA) were analyzed in cell-culture medium by the corresponding kits. RT-qPCR was used to detect the mRNA expression of APP, BACE-1 and GSK-3β . RESULTS Compared with the model group, the 50%, 70%, 95% concentration of alcohol in n-butanol components of semen euryales improved the cell viability of PC12(P<0.05). The 70% alcohol components significantly increased SOD activity and decreased the content of MDA in culture supernatants (P<0.05) and significantly down-regulated the expression of GSK-3β (P<0.01). CONCLUSION The 70% and 95% alcohol components of n-butanol extracts of semen euryales shows significant protective effects on oxidative stress-induced cell injury. The protection mechanism may be related to the increased SOD activity, the reduced the emission of MDA, and the inhibition the key genes associated with the accumulation of Aβ.

     

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