丹酚酸B通过SIRT1/PGC-1α通路对Aβ_(1-42)干预N2A细胞保护作用研究

Salvianolic acid B protects against Aβ-induced neuronal injury via repressing oxidative stress and SIRT1/PGC-1αsignaling

目的观察沉默信息调节因子2相关酶1(SIRT1)/过氧化物酶体增殖物激活受体γ共激活因子1α(PGC-1α)的表达及检测活性氧(ROS)、丙二醛(MDA)、超氧化物歧化酶(SOD)含量和线粒体膜电势,探讨丹酚酸B(SalB)减轻β淀粉样多肽1-42(Aβ1-42)干预小鼠来源神经瘤母细胞(N2A)后氧化应激损伤的作用及机制。方法使用10μM Aβ1-42寡聚体干预N2A细胞构建阿尔茨海默病(AD)细胞模型,使用40μM SalB干预细胞为对照组,模型组和SalB干预组。使用MTT法检测不同实验组细胞活力;DCFH-DA染色测定实验组细胞内ROS水平;ELISA法检测SOD,MDA水平;Western blot法和RTPCR法分别检测不同实验组SIRT1、PGC-1α蛋白和mRNA水平。结果与Aβ干预N2A细胞构建的模型组相比,SalB组处理后的模型组细胞活力显著升高(P<0.001),SalB组细胞中ROS水平显著下降(P<0.01),SOD水平显著上升(P<0.001),MDA生成显著减少(P<0.05),有效恢复线粒体膜电势(P<0.05)。另外,SalB处理后模型组细胞的SIRT1、PGC-1α蛋白和mRNA水平均升高。结论SalB可以显著降低Aβ干预N2A细胞后诱导的氧化应激反应,减少ROS产生及下调MDA水平,上调SOD水平,该神经保护作用可能与上调SIRT1/PGC-1α通路相关。

Objective To observe the expression of sirtuin 1(SIRT1)/peroxisome peroxisome proliferatoractivated receptor-gamma coactivator 1 alpha(PGC-1α),and to investigate the effect and mechanism of salvianolic acid B(SalB)on reducing oxidative stress injury after Aβ intervention in neuroblastoma(N2A)cells.Methods N2A cells were treated with 10μM Aβ oligomers to construct Alzheimer's disease(AD)cell models.After 40μM SalB intervention,N2A cells were divided into control group,Aβ treated group and SalB+Aβ treated group.MTT assay was used to detect cell viability and the intracellular reactive oxygen species(ROS)levels were determined by DCFH-DA staining.The SOD and MDA levels were detected by ELISA kits.The protein and mRNA levels of SIRT1,PGC-1α in different experimental groups were detected by Western blot and RT-PCR.Results Compared with the Aβ treated group,the cell vitality of N2A cells in SalB+Aβ treated group was increased(P<0.001),the ROS level in SalB group was decreased significantly(P<0.01),SOD level was increased significantly(P<0.01),MDA production was decreased significantly(P<0.05).In addition,the protein and mRNA levels of SIRT1 and PGC-1α was upregulated significantly after SalB treatment.Conclusions SalB can significantly ameliorate Aβ induced oxidative stress by reducing ROS production and down-regulating the levels of SOD and MDA,which may be associated with its regulation of SIRT1/PGC-1α expression.