松果菊苷对帕金森病模型小鼠黑质纹状体蛋白表达影响的双向电泳分析  

Effects of echinacoside on protein expression from substantia nigra and striatal tissue in mouse MPTP model of Parkinsons disease by using 2-dimensional electrophoresis analysis

作　　者：赵欣[1] 蒲小平[1] 耿兴超[1] 

机构地区：[1]北京大学药学院分子与细胞药理学系

出　　处：《中国药理学通报》2008年第1期28-32,共5页Chinese Pharmacological Bulletin

基　　金：国家重点基础研究发展计划(973计划)资助项目(No2004CB518902);国家自然科学基金资助项目(No30472164)

摘　　要：目的观察松果菊苷对MPTP致小鼠帕金森病(PD)模型中对动物行为学和对黑质纹状体组织蛋白表达的影响,在蛋白质水平探讨松果菊苷的多巴胺能神经元保护作用机制。方法以MPTP损伤建立C57BL/6小鼠PD模型,通过自主活动实验和滚筒实验观察动物行为学表现,用双向电泳、图像分析、质谱鉴定等蛋白质组学技术研究松果菊苷对MPTP致小鼠PD模型行为学的影响和黑质纹状体组织蛋白表达的变化。结果①经MPTP诱导,小鼠的自主活动次数、滚筒运动潜伏期均低于对照组(P<0.01);经松果菊苷(20mg.kg-1)预处理后,MPTP诱导小鼠的行为学表现明显改善(P<0.01)。②经过双向电泳及PDQuest软件分析,硝酸银染色,对照组、模型组和松果菊苷给药组分别分离出282个、269个和236个的蛋白质斑点。对一个稳定差异表达的斑点进行MALDI-TOF质谱鉴定,数据库检索结果为胆绿素还原酶B。结论松果菊苷对MPTP致小鼠PD模型的行为学障碍具有改善作用,其神经保护作用机制可能与胆绿素还原酶B水平降低有关。Ahn To study the effect of echinacoside on behavior and proteins expression from substantia nigra and striatal tissue in MPTP mouse model of Parkinsons disease（PD） and discover the mechanism of its potential dopaminergic neuroprotective effect in the protein level. Methods The mouse model of PD was induced by 1-Methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine （MPTP）and the behavioral analysis of C57BL/6 mice was performed by using spontaneous movement and rotarod test. A proteomic approach based on 2-dimen- sional electrophoresis （ 2-DE ）, mass spectrometry （MS） and figure analysis was used to evaluate the effect of echinacoside on the behavior and the protein expression in substantia nigra and striatal tissue in C57BL/6 mice after MPTP administration. Results ① Compared with control, MPTP lesion significantly reduced the number of spontaneous movement and latent period of mice on the rotating rod （ both P 〈 0.01 ）. Compared with MPTP model group, the number ofspontaneous movement and latent period of mice on the rotating rod were significantly increased（ both P 〈0. 01 ） in echinacoside-pretreated groups. ②Proteins extracted from striatal and substantia nigra tissue of the control, model and echinacoside treated mice were resolved in parallel on a 2-D gel and visualized by silver staining. The images were analyzed by using PDQuest software. Every specimen contained a total of more than 200 spots. Among the protein spots with significantly changed staining intensity, a spot was selected for MS analysis. It was identified as biliverdin reductase B. Conclusions Echinacoside might be able to protect C57BL/6 mice against MPTP-induced behavioral default. The neuroprotective effect might be associated, at least partially, with decreasing the biliverdin reductase B level

关 键 词：松果菊苷 MPTP 帕金森病 双向凝胶电泳 胆绿素还原酶B 

分 类 号：R-332[医药卫生] R284.1