芡实正丁醇部位分离组分对H_2O_2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 H_2O_2-Induced PC12 Cells

目的研究芡实正丁醇部位的聚酰胺柱分离组分对H_2O_2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200μmol/L的H_2O_2损伤细胞8h,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-3βmRNA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。

OBJECTIVE To investigate the effect of n-butanol extract fromEuryale feroxSalisb.against H_2O_2-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 200μmol/L H_2O_2 for 8h 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-1and 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.