丁苯酞对缺氧致PC12细胞损伤保护作用的比较蛋白质组研究  

Comparative proteomics study of dl-3-n-butylphthalide protecting PC12 cells from hypoxiainduced damage

作　　者：杨国锋[1] 高雅[1] 王伟 纪建国[3] 李小芳[1] 张锐利[1] 许蕾[1] 

机构地区：[1]河北医科大学第二医院神经科,河北石家庄050000 [2]总后勤部管理局北京金沟河干休所卫生所,北京100853 [3]北京大学生命科学学院蛋白质工程国家重点实验室,北京100871

出　　处：《中华神经医学杂志》2013年第3期251-255,共5页Chinese Journal of Neuromedicine

摘　　要：目的从蛋白质水平研究丁苯酞对连二亚硫酸钠（Na2S2O4）致PCI2细胞缺氧的保护作用及其作用机制。方法利用含Na2S2O4的无血清培养基处理PCI2细胞，建立细胞缺氧模型，并观察丁苯酞对细胞缺氧的保护作用。噻唑盐比色法（MTT）测定细胞活性。蛋白质组学技术鉴定差异表达蛋白。结果随着丁苯酞浓度的增大，细胞平均吸光度似）值逐渐增加。当丁苯酞浓度大于等于5mol／L时，细胞平均A值与未添加丁苯酞的模型组细胞A值差异有统计学意义似0．05）。蛋白质组学技术鉴定出PCI2细胞缺氧模型组与丁苯酞干预组共17个差异表达蛋白点，多为细胞骨架蛋白、凋亡相关蛋白及氧化应激相关蛋白等。结论丁苯酞可减少缺氧导致的神经元凋亡。细胞骨架蛋白、凋亡相关蛋白及氧化应激相关蛋白可能在丁苯酞的神经保护机制中发挥作用。Objective To explore the protective effect and molecular mechanism of dl-3-n-butylphthalide （NBP） against oxygen-glucose deprivation induced by sodium hydrosulfite in PC12 cells at proteomic level. Methods PC12 cells impaired by serum-free culture media with sodium hydrosulfite were used as the cell models ofhypoxia, and the protective effects of NBP on hypoxic cells were observed. Methyl thiazolyl tetrazolium （MTT） assay was used to measure the viabilities of PC12 cells. Proteomic technique was employed to identify the differential expression proteins. Results Following the increment of NBP concentrations, the cell absorbance （A） value increased gradually （the PC12 cell apoptosis reduced gradually）; when the NBP concentration reached 5 mol/L, their cell A value was significantly different as compared with that of cells without adding NBP （P〈0.05）. By using proteomics methods, 17 differentially-expressed protein spots in the cells without adding NBP and cells with NBP were identified; most of proteins were cytoskeleton proteins, apoptosis related proteins and oxidative stress related proteins. Conclusion NBP can reduce neuronal apoptosis induced by hypoxia, and cytoskeletal proteins, apoptosis-related proteins and oxidative stress related proteins may role in protection of NBP on ischemic neurons.

关 键 词：丁苯酞 PC12细胞 缺氧 连二亚硫酸钠 蛋白质组学 

分 类 号：R285.5[医药卫生—中药学]