Aβ_(25-35)诱导PC12细胞损伤的蛋白质组学研究  

Proteomic studies of-amyloid_((25-35))-treated PC12 cells

作　　者：李红杰[1] 胡林森[2] 常明[2] 程永杰[1] 张磊[3] 侯澍[4] 孙肖爽[1] 

机构地区：[1]长春市中心医院神经内科,吉林长春130051 [2]吉林大学第一医院神经内科,吉林长春130021 [3]吉林大学第二临床学院儿科,吉林长春130041 [4]北京世纪坛医院神经内科,北京100038

出　　处：《中风与神经疾病杂志》2009年第5期527-531,共5页Journal of Apoplexy and Nervous Diseases

基　　金：吉林省卫生厅科研立项课题(No.2008Z075)

摘　　要：目的识别AD细胞模型中蛋白质组改变,在蛋白质水平上揭示Aβ的作用机制。方法利用双向差异凝胶电泳技术(2D-DIGE)和质谱(MS)技术,探索20μmol/L浓度Aβ25-35肽段作用48h后PC12细胞蛋白质组的改变。结果2D-DIGE图像上出现约2000个蛋白点。与对照组比较,Aβ25-35作用下共有29个蛋白的表达有显著差异,其中25个蛋白表达量上调及4个蛋白表达量下调超过30%。质谱鉴定出7个蛋白点,分为3类:(1)具有分子伴侣活性的蛋白:葡萄糖调节蛋白75(glucose-regulated protein75,GRP75)、热休克同源蛋白71(heat shock cognate71kDa protein,HSC71)、calreticulin表达量均上调。(2)细胞骨架蛋白:β-微管蛋白(tubulin beta chain15,TBETA-15)和低分子量神经细丝蛋白(neurofilament light polypeptide,NF-L)表达量亦上调。(3)与能量代谢有关的酶:肌酸激酶B(creatine kinase-B,CKB)和醛缩酶A(aldolaseA)蛋白表达量均下调。结论本实验首次将DIGE和MS方法应用于Aβ25-35神经毒性作用机制研究,在蛋白质组水平上揭示了Aβ25-35毒性作用的早期机制。Objective Explore the toxic mechanisms of Aβ at proteomic level. Methods 2D-DIGE and MALDI- TOF MS technology were used to detect proteomic changes of PC12 cells treated with Aβ（25-35）. Results About 2000 protein spots were seen in 2D-DIGE images. The expressions of 29 proteins were significantly changed, of which 25 were up-regulated and 4 were down-regulated more than 30% in Aβ-treated PC12 ceils compared with control ones. 7 protein spots were identified by mass spectrometry. They were primarily divided into 3 categories： （ 1 ） proteins with chaperone activity ： glucose-regulated protein75, heat shock cognate 71 kDa protein, and calreticulin, which were all up-regulated. （2） Cytoskeletal proteins, tubulin and neurofilament light polypeptide were up-regulated. （3）Energy-related enzyme, creatine kinase B and aldolase A were down-regulated. Conclusion For the first time,the toxic mechanisms of Aβ were studied by DIGE and MS technology at proteomic level. Our proteomic results provide new clues for future studies of AD.

关 键 词：阿尔茨海默病 AΒ25-35 蛋白质组学 PC12细胞 双向差异凝胶电泳 质谱 

分 类 号：R749.16[医药卫生—神经病学与精神病学] Q51[医药卫生—临床医学]