柴胡皂苷D对铝暴露条件下人神经元细胞磷酸化Tau蛋白水平的影响

Saikosaponin D Inhibiting Over-phosphorylation of Tau Protein in Human Neuron Cells in Aluminum Exposure Condition

目的:通过观察柴胡皂苷D(SSD)对铝诱导人神经元细胞磷酸化Tau蛋白表达的影响,探讨柴胡皂苷D防治AD的可能机制。方法:选取人神经母细胞瘤细胞系SH-SY5Y细胞,培养基中加入神经生长因子(NGF)诱导SHSY5Y细胞分化成神经元细胞后,将细胞分为5组:对照组,常规培养基培养;Al Cl3处理组,用含有100μmol/L Al Cl3的常规培养基培养;SSD处理组,在含有100μmol/L Al Cl3的常规培养基中分别加入终浓度为1μg/m L、2μg/m L和4μg/m L SSD。各组细胞培养48 h后,进行实验分析。四甲基偶氮唑蓝(MTT)法检测细胞活力;Western Blot方法检测各组细胞磷酸化Tau蛋白表达情况。结果:与对照组相比较,100μmol/L Al Cl3对细胞的活力无明显影响(P>0.05);不同浓度的SSD对细胞活力无显著影响(P>0.05);与对照组相比,铝暴露组细胞中磷酸化Tau蛋白明显增高(P<0.05),在加入不同浓度的SSD后,与铝暴露组细胞相比较,磷酸化Tau蛋白水平明显降低(P<0.05),并呈剂量依赖关系。结论:SSD可以抑制铝暴露诱导的神经元细胞Tau蛋白过度磷酸化。

Objective： By observing the effects of saikosaponin D（SSD） on expressions of Tau protein in neuronal cells in aluminum exposure condition,to explore the possible mechanism by which saikosaponin D may function as a positive role in the prevention of Alzheimer＇s disease. Methods： The human neuroblastoma cell line SH-SY5 Y cells were used to differentiate into neuronal cells by adding nerve growth factor（NGF） in the culture medium. The cells were divided into 5 groups： control group in which the cells were cultured in conventional medium culture; Al Cl3 group in which the cells were cultured in conventional medium culture containing 100 μmol / L Al Cl3; SSD groups in which SSD was added in a final concentration of 1 μg/m L,2 μg/m L and4 μg / m L in the medium containing 100 μmol / L Al Cl3. After 48 h,experimental analysis was carried out. Tetrazolium blue（MTT） assay was used to determine the cell viability. Western blot method was used to detect the intracellular phosphorylation of Tau protein. Results： Compared with the control group,100 μmol / L Al Cl3 had no significant effect（P〉0. 05） on the viability of the cells. The different concentrations of SSD also had no significant effect（P〉0. 05） on cell viability. Compared with the control group,the phosphorylation of Tau protein was significantly increased in Al-exposed group（P〈0. 05）. After the interference with different concentrations of SSD,compared with aluminum group,levels of phosphorylation of Tau protein were significantly lower（P〈0. 05） in a dose-dependent manner. Conclusion： SSD can inhibit hyperphosphorylation of Tau protein in neuronal cells induced by aluminum exposure.