摘要
目的探讨毛蕊异黄酮对H2O2刺激的SY5Y细胞氧化应激损伤的调控作用及其机制。方法MTT法检测毛蕊异黄酮对SY5Y细胞活性的影响以筛选合适的药物干预浓度;实验分PBS对照组,H2O2(250μmol/L)模型组和H2O2(250μmol/L)+毛蕊异黄酮(0.035μmol/mL)干预组;试剂盒检测MDA的含量和SOD的活性;免疫荧光染色检测Nrf2的核内转移情况及HO-1的表达,免疫印迹法检测总Nrf2、核内Nrf2、NQO1和HO-1蛋白水平。结果与PBS对照组比较,H2O2组SY5Y细胞活性降低,毛蕊异黄酮可以改善H2O2诱导的细胞损伤,降低MDA的含量、增加SOD的活性。同时,毛蕊异黄酮能够激活Nrf2并促进其向核内转位,明显上调H2O2损伤后细胞总Nrf2、核内Nrf2、NQO1和HO-1的水平。结论毛蕊异黄酮可能通过激活Nrf2/HO-1信号通路抑制H2O2诱导的SY5Y细胞氧化应激损伤。
Objective To explore the regulatory effect and mechanism of calycosin on oxidative stress injury of SY5Y cells induced by H2O2.Methods MTT assays were used to assess the effect of calycosin on the activity of SY5Y cells to screen for the appropriate drug concentration;The experiment included a PBS control group,H2O2(250μmol/L)model group,and H2O2(250μmol/L)+calycosin(0.035μmol/mL)intervention group.Kits were used to detect MDA content and SOD activity.Immunofluorescence staining was used to detect nuclear transfer of Nrf2 and expression of HO-1.Western blotting detected the levels of total Nrf2,nuclear Nrf2,NQO1,and HO-1 proteins.Results Compared with the PBS control group,the activity of SY5Y cells in the H2O2 group was reduced and calycosin improved H2O2-induced cell damage,reduced the level of MDA,and increased the activity of SOD.Calycosin also activated Nrf2 and promote its translocation into the nucleus,which significantly upregulated the levels of total Nrf2,nuclear Nrf2,NQO1,and HO-1 after H2O2 injury.Conclusions Calycosin inhibited H2O2-induced oxidative stress injury in SY5Y cells by activating Nrf2/HO-1 signaling.
作者
白振军
张慧宇
郭敏芳
BAI Zhenjun;ZHANG Huiyu;GUO Minfang(College of Traditional Chinese Medicine Health Service,Shanxi Datong University,Datong 037009,China;Medical College,Shanxi Datong University,Datong 037009)
出处
《中国比较医学杂志》
CAS
北大核心
2020年第10期37-43,共7页
Chinese Journal of Comparative Medicine
基金
山西省平台基地专项(201805D111009)
山西省高等学校科技创新项目(2020L0484)
2019年神经炎症及变性疾病基础与应用研究山西省重点实验室开放课题(KF2019007)。
