基于Nrf2/HO-1通路的黄芪-灯盏细辛成分配伍对缺氧缺糖PC12细胞氧化损伤研究  

Effect of Combined Components of Huangqi(黄芪)and Dengzhanxixin(灯盏细辛)on the Oxidative Damage in PC12 Cells Induced by Oxygen-Glucose Deprivation through Nrf2/HO-1 Signaling Pathway

作　　者：鄢宇梅 米佳佳 付英豪 程芳 张洁 曾瑾 盛艳梅[1] Yan Yumei;Mi Jiajia;Fu Yinghao;Cheng Fang;Zhang Jie;Zeng Jin;Sheng Yanmei(School of Pharmacology,Chengdu Medical College,Chengdu 610500;Sichuan Academy of Chinese Medicine Sciences,Chengdu 610000)

机构地区：[1]成都医学院药学院,成都610500 [2]四川省中医科学院,成都610000

出　　处：《中药药理与临床》2022年第2期159-164,共6页Pharmacology and Clinics of Chinese Materia Medica

基　　金：四川省科技厅重点研发项目(编号:2020YFS0325);四川省杰出青年科技人才项目(编号:2020JDJQ0063)

摘　　要：目的:探索黄芪与灯盏细辛中有效成分黄芪甲苷、绿原酸、灯盏乙素最佳配比组合及其对缺糖缺氧PC12细胞氧化损伤的保护作用。方法:采用PC12细胞株、无糖DMEM培养基和氯化钴构建慢性脑缺血细胞模型。采用均匀设计法筛选黄芪甲苷、绿原酸、灯盏乙素最优配比组合,并采用流式细胞术检测活性氧(ROS)含量,罗丹明123检测线粒体膜电位,WB技术检测Nrf2、HO-1蛋白表达,评价最优组合的作用机制。结果:0.4 mmol/L的氯化钴为PC12缺糖缺氧造模条件。3个单体最优组合为黄芪甲苷∶绿原酸∶灯盏乙素=10∶75∶40时显著提高PC12细胞存活率(P<0.01);与模型对照组比,最优组合的活性氧含量显著降低(P<0.01),线粒体膜电位显著升高(P<0.01),Nrf2和HO-1蛋白表达明显上调(P<0.05)。结论:黄芪甲苷∶绿原酸∶灯盏乙素=10∶75∶40是最优组合,该组合通过降低细胞内活性氧含量并稳定线粒体膜电位促进缺血缺氧PC12细胞存活,上调Nrf2、HO-1蛋白表达,对慢性脑缺血性氧化损伤产生保护作用。Objective:To explore the optimal combination of astragaloside,chlorogenic acid,and scutellarin,the active components in Huangqi(黄芪)and Dengzhanxixin(灯盏细辛),against oxidative damage in PC12 cells with oxygen-glucose deprivation(OGD).Methods:Chronic cerebral ischemia was induced in PC12 cells with glucose-free DMEM and cobalt chloride.The uniform design method was adopt to screen the optimal combination of astragaloside,chlorogenic acid,and scutellarin,flow cytometry to detect the content of reactive oxygen species(ROS),rhodamine 123 to determine mitochondrial membrane potential(MMP),and Western blotting(WB)to detect the expression of Nrf2 and HO-1.Thereby,mechanism of the optimal combination was clarified.Results:The 0.4 mmol/L cobalt chloride was finally used to induce OGD in PC12.Astragaloside,chlorogenic acid,and scutellarin at the optimal ratio of 10:75:40 significantly improved PC12 cell survival(P<0.01).Compared with the model group,the optimal combination decreased the content of ROS(P<0.01),increased MMP(P<0.01),and raised the expression of Nrf2 and HO-1(P<0.05)of the cells.Conclusion:Astragaloside,chlorogenic acid,and scutellarin at the ratio of 10:75:40 demonstrate the best effect,which promote the survival of OGD PC12 cells by reducing ROS and stabilizing MMP and protect the cells against oxidative damage by up-regulating the expression of Nrf2 and HO-1.

关 键 词：黄芪-灯盏细辛成分配伍 慢性脑缺血 氧化损伤 PC12细胞 均匀设计法 

分 类 号：R285[医药卫生—中药学]