黄芪甲苷对H_2O_2致PC12细胞氧化应激损伤的保护作用  

A study on protection of astragaioside Ⅳ about oxidative stress on PC12 cells induced by H_2O_2

作　　者：王世博[1] 邱景富[2] 白群华[2] 李佳佳[1] 和晋渝[2] 高艳军[3] 于超[1] 

机构地区：[1]重庆医科大学生命科学研究院,重庆400016 [2]重庆医科大学公共卫生学院,重庆400016 [3]重庆医科大学检验医学院,重庆400016

出　　处：《中国药理学通报》2011年第11期1603-1609,共7页Chinese Pharmacological Bulletin

基　　金：重庆市自然科学基金资助项目(NoCSTC;2009BA5083)

摘　　要：目的探讨黄芪甲苷对H2O2引起PC12细胞氧化应激损伤的保护作用与分子机制。方法用H2O2作用PC12细胞建立氧化应激损伤模型,通过MTT法检测细胞活力;Hoechst 33258染色观察细胞内核酸形态变化;流式细胞术检测细胞凋亡、胞内活性氧的产生及细胞周期变化情况;Western blot检测细胞周期蛋白Cyclin D1、Cyclin A、磷酸化p38及T-p38的表达。结果成功建立了H2O2致PC12细胞氧化应激损伤模型;黄芪甲苷能够提高H2O2损伤的PC12细胞的活力,提升因H2O2导致的细胞凋亡率的下降,降低H2O2所致的胞内ROS升高的状况。机制研究表明:黄芪甲苷通过恢复H2O2诱导细胞周期蛋白Cyclin D1表达下调的情况,调控各细胞周期的百分比,恢复细胞的正常增殖;进一步研究发现黄芪甲苷是通过抑制H2O2对p38的激活发挥作用的。结论黄芪甲苷对H2O2致PC12细胞氧化应激损伤具有保护作用,其作用机制可能是通过调控细胞周期蛋白Cyclin D1的表达来实现的,调控过程与p38/MAPK通路密切相关。该发现为临床上抗氧化治疗策略提供有益的实验依据。Aim To investigate the effect of astragaioside Ⅳ on PC12 cell injury induced by H2O2 and its underlying mechanism.Methods The viability of PC12 cells induced by H2O2 was detected by MTT assay.The status of nucleic acid in cell was observed by Hoechst 33258 staining.Apoptosis of PCI2 cells,reactive oxygen species（ROS） and cell cycle were measured by flow cytometry.The expression of Cyclin D1,Cyclin A,Phospho-p38 and T-p38 was examined by Western blot.Results The model on oxidative stress of PC12 cells induced by H2O2 was constructed successfully.The cell viability of PC12 injured by H2O2 was rescued by astragaioside Ⅳ,which could inhibit apoptosis of PCI2 cells and reduce ROS and increase percentage of G1-phase in cell cycle.Furthermore,astragaioside Ⅳ could rescue the descending expression of Cyclin D1 and the increasing expression of Phospho-p38 induced by H2O2.Conclusions Astragaioside Ⅳ protects PC12 cells from oxidative stress induced by H2O2 via stimulating the expression of Cyclin D1 and inhibiting active Phospho-p38.The present data suggest that Cyclin D1 and Phospho-p38 might be a target of clinical therapy via antioxidation.It is an insight into prevention of neurodegenerative diseases.

关 键 词：黄芪甲苷 过氧化氢 氧化应激 CYCLIN D1 CYCLIN A PC12细胞 

分 类 号：R284.1[医药卫生—中药学] R329.24[医药卫生—中医学]