新型双香豆素类化合物通过Nrf 2/ARE通路诱导OGD神经元保护研究

A new biscoumarin compound protects against neuronal oxygen and glucose deprivation via Nrf 2/ARE pathway

目的:探讨新型双香豆素类化合物3-CF3-4-CL对氧糖剥夺损伤(OGD)神经元诱发氧化应激的影响和潜在作用机制。方法:原代培养小鼠皮层神经元,建立OGD模型模拟脑缺血再灌注损伤;以Cell Counting Kit-8(CCK-8)法检测3-CF3-4-CL对细胞生存率的影响;q-PCR检测四种抗氧化因子的mRNA表达;以丙二醛(MDA)、超氧化物歧化酶(SOD)和谷胱甘肽过氧化物酶(GSH-Px)为细胞氧化水平指标;Western blot检测Nrf 2/ARE信号通路蛋白水平变化。结果:与对照组相比,3-CF3-4-CL处理组细胞生存率提高,抗氧化因子mRNA表达量升高,SOD和GSH-Px表达水平升高,MDA含量降低;3-CF3-4-CL处理组细胞核因子E2相关因子2(Nrf 2)总蛋白活化水平升高,细胞核Nrf 2蛋白水平表达升高;同时,其下游抗氧化蛋白血红素氧合酶-1(HO-1)表达水平升高。结论:新型双香豆素类化合物3-CF3-4-CL通过激活Nrf 2/ARE信号通路,提高OGD损伤神经元的抗氧化应激水平,发挥神经保护功能。

Objective:To investigate the effect and potential mechanism of a new type biscoumarin compound named 3-CF3-4-CL,during protection against oxidative stress caused by Oxygen and glucose deprivation(OGD)in neurons.Methods:The in vitro model to assess the mechanism of oxidative stress in I/R was established by OGD of neurons.The cell viability was measured by CCK-8 assay.The mRNA levels of four antioxidant elements was measured by q-PCR.Oxidative stress indicators was detected by malondialdehyde(MDA),superoxide dismutase(SOD),glutathione peroxidase(GSH-Px).The effects of 3-CF3-4-CL on nuclear translocation of Nrf 2 and protein expression of hemeoxygenase-1(HO-1)were measured by western blot.Results:Compared with the control group,in 3-CF3-4-CL treated group the cell viability of neurons increased obviously,the mRNA of four antioxidant elements increased,3-CF3-4-CL improved SOD and GSH-Px activation,reduced MDA content.Moreover,3-CF3-4-CL could induce activation and nuclear translocation of nuclear factor E2 related factor 2(Nrf 2)and significantly increased protein expression of HO-1 when compared with OGD group.Conclusion:3-CF3-4-CL may protect against damage of oxidative stress in OGD model of neurons via activation of Nrf 2/ARE pathway.