静宁颗粒对H2O2诱导的原代皮层神经元氧化应激的保护作用研究

目的:探讨静宁颗粒对小鼠原代皮层神经元氧化损伤的保护作用。方法:实验分为正常组、模型组和各用药组。采用过氧化氢(H₂O₂)刺激造成原代皮层神经元氧化损伤模型,各用药组采用0.2、0.4、0.8 mg/ml的静宁颗粒进行干预。采用MTT法检测活性抑制率,采用试剂盒检测活性氧(ROS)、丙二醛(MDA)、谷胱甘肽(GSH)、超氧化物歧化酶(SOD)活力; Western blot检测kelch样ECH关联蛋白1(Keap1)、红系衍生的核转录相关因子2(Nrf2)、促凋亡蛋白(Bax)、B淋巴细胞瘤-2基因(Bcl-2)及含半胱氨酸的天冬氨酸蛋白水解酶3(Caspase 3)等蛋白的表达。结果:静宁颗粒干预后较模型组相比ROS、MDA降低,GSH、SOD升高; 与模型组比较,Keap1、p53、Bax及Caspase 3蛋白表达下调,Nrf2、NQO1、HO-1及Bcl-2蛋白表达升高。结论:静宁颗粒可通过Keap1/Nrf2氧化应激通路和Caspase依赖的线粒体通路对H₂O₂所致小鼠原代皮层神经元氧化应激损伤发挥保护作用。

Protective effect of Jingning granule on H2 O2 induced oxidative stress in primary cortical neurons

Objective:To investigate the protective effect of Jingning granule on oxidative damage of primary cortical neurons extracted from mice.Methods:The experiment was divided into normal group,model group and administration group.The oxidative damage model of primary cortical neurons caused by hydrogen peroxide(H₂O₂)stimulation was used.The administration group was intervened with Jingning granules of 0.2,0.4,0.8 mg/ml.MTT assay was used to detect the inhibition rate.The kits were used to detect the activity of ROS,MDA,GSH and SOD;the protein expression of Kelch-like ECH associated protein1(Keap1),Nuclear factor(erythroid-derived2)like 2 protein(Nrf2),Bax,B-cell lymphoma-2(Bcl-2)and cysteinyl aspartate specific proteinase(Caspase 3)is detected by Western blot.Results:Compared with the model group,Jingning granule decreased ROS,MDA,and increased GSH,SOD.The expression of Keap1,p53,Bax and Caspase 3 protein was down-regulated,and Nrf2,NQO1,HO-1 and Bcl-2 was up-regulated compared with the model group.Conclusion:Jingning granule can protect primary cortical neurons from oxidative stress damage induced by H₂O₂ through Keap1/Nrf2 oxidative stress pathway and Caspase dependent mitochondrial pathway.