摘要
目的探讨原花青素B2(proanthocyanidins B2,PCB2)对过氧化氢(H_(2)O_(2))诱导的人少突胶质细胞(MO3.13)氧化损伤和凋亡的保护作用及其机制。方法筛选H_(2)O_(2)和PCB2的最佳作用浓度。分为正常组、PCB2组(100 mg·L^(-1) PCB2处理24 h)、H_(2)O_(2)模型组(500μmol·L^(-1) H_(2)O_(2)处理24 h)、H_(2)O_(2)+PCB2组(500μmol·L^(-1) H_(2)O_(2)与100 mg·L^(-1) PCB2共同处理24 h)。FRAP法检测PCB2的抗氧化能力;CCK-8法检测各组细胞存活率,LDH法进行细胞毒性检测;微量酶标法和ELISA法检测各组细胞中LDH、NO、H_(2)O_(2)含量以及CAT、SOD活力;免疫荧光和Western blot分别检测各组细胞中NRF2、xCT、HO-1、Ferritin、GPX4的蛋白表达水平。亚铁离子荧光探针(FerroOrange)检测细胞内亚铁离子(Fe^(2+))含量。结果H_(2)O_(2)能诱导MO3.13氧化损伤并导致细胞铁死亡,PCB2能够减轻MO3.13氧化损伤和铁死亡;与H_(2)O_(2)模型组相比,PCB2干预能够明显升高MO3.13内LDH含量,降低NO、H_(2)O_(2)含量,提高SOD、CAT活力;上调NRF2、xCT、HO-1、Ferritin、GPX4的蛋白表达水平。结论PCB2能够通过NRF2/HO-1/xCT/GPX4轴增强细胞抗氧化能力,减轻H_(2)O_(2)诱导的MO3.13氧化损伤。
Aim To explore the protective effect of anthocyanin B2(PCB2)on hydrogen peroxide(H_(2)O_(2))induced oxidative damage and apoptosis in human oligodendrocytes(MO3.13)and the underlying mechanism.Methods The optimal concentration of H_(2)O_(2) and PCB2 for action was screened,and divided into normal group,PCB2 group(100 mg·L^(-1) PCB2 treatment for 24 hours),H_(2)O_(2) model group(500μmol·L^(-1) H_(2)O_(2) treatment for 24 hours),and H_(2)O_(2)+PCB2 group(500μmol·L^(-1) H_(2)O_(2) and 100 mg·L^(-1) PCB2 co-treated for 24 hours).FRAP method was used to detect the antioxidant capacity of PCB2;CCK-8 method was used to detect the survival rate of cells in each group,while LDH method was used to assess cytotoxicity.Microenzyme-linked immunosorbent assay and ELISA were used to examine the levels of LDH,NO,H_(2)O_(2),as well as the activities of CAT and SOD in each group of cells.Immunofluorescence and Western blot were used to detect the protein expression levels of NRF2,xCT,HO-1,ferritin,and GPX4 in each group of cells.FerroOrange fluorescent probe was used to detect the intracellular content of ferrous ions(Fe^(2+)).Results H_(2)O_(2) could induce MO3.13 oxidative damage and lead to cell ferroptosis,while PCB2 could alleviate MO3.13 oxidative damage and ferroptosis.Compared with the H_(2)O_(2) model group,PCB2 intervention could significantly increase LDH content in MO3.13,reduce NO and H_(2)O_(2) content,and improve SOD and CAT activity,and up-regulate the protein expression levels of NRF2,xCT,HO-1,ferritin,and GPX4.Conclusion PCB2 can enhance cellular antioxidant capacity and alleviate H_(2)O_(2) induced MO3.13 oxidative damage through the NRF2/HO-1/xCT/GPX4 axis.
作者
刘健
陈莹
梁亚杰
蒲萌
张紫薇
郑璐璐
柴智
肖莹
马存根
王青
LIU Jian;CHEN Ying;LIANG Ya-jie;PU Meng;ZHANG Zi-wei;ZHENG Lu-lu;CHAI Zhi;XIAO Ying;MA Cun-gen;WANG Qing(The Key Research Laboratory of Benefiting Qi for Acting Blood Circulation Method to Treat Multiple Sclerosis of State Administration of Traditional Chinese Medicine,Research Center of Neurobiology,Shanxi University of Chinese Medicine,Jinzhong Shanxi 030619,China;Wuhan Caidian District People's Hospital,Wuhan 430100,China)
出处
《中国药理学通报》
CAS
CSCD
北大核心
2024年第9期1735-1743,共9页
Chinese Pharmacological Bulletin
基金
国家青年科学基金资助项目(No 81903596)
山西省回国留学人员科研资助项目(No 2022-165)
国家中医药管理局科研课题(No 2023ZYYDA2038)
山西省卫健委医学科技领军团队(No 2020TD05)
山西省中医药管理局科研课题(No 2023ZYYB040)
山西省卫健委2022年度中医药科研课题立项计划(No 2022ZYYC090)
山西中医药大学学科建设经费(No 2024XKJS-02)
山西省教育厅科技创新项目(No 2022L336)
山西中医药大学2022年度科技创新团队(No 2022TD2006)
山西中医药大学科技创新能力培育计划(No 2022-TY-PH-11,2022-TY-PH-37)。
