人体失神经萎缩骨骼肌蛋白质组学的初步研究  

The proteomics study of denervated human skeletal muscles

作　　者：徐雷[1] 顾玉东[1] 徐建光[1] 贾林芝[2] 张志鸿[2] 

机构地区：[1]复旦大学附属华山医院手外科,上海200040 [2]复旦大学生命科学学院

出　　处：《中华手外科杂志》2005年第4期236-240,共5页Chinese Journal of Hand Surgery

基　　金：中国博士后基金资助项目(2003034295);复旦大学Med-X项目;教育部重大课题项目(0103);上海市科委重大项目(04DZ19901)

摘　　要：目的采用蛋白质组学分析方法研究人类失神经萎缩骨骼肌肌细胞的蛋白质组表达变化及其临床意义.方法以臂丛神经根性撕脱伤患者为对象,对失神经萎缩的肱二头肌、小鱼际肌及其健侧胸锁乳突肌肌纤维组织中的蛋白质进行二维电泳和PD-Quest分析,对差异蛋白质用MALDI-TOF质谱仪进行Trypsin酶解指纹图测定.结果(1)初步建立了人体骨骼肌蛋白质组学研究的技术平台,建立了失神经萎缩肱二头肌、小鱼际肌以及健侧胸锁乳突肌的凝胶图像的Matchset;(2)在失神经萎缩肌肉中800个蛋白质共有32个发生了变化,表达变化蛋白质主要集中在pI 4～8,Mr 相对分子质量为30000～50000范围内;(3)发现了肌肉萎缩部分的可能相关蛋白,在萎缩组共有6个点的表达量增加,24个点的表达量降低,两者差异有统计学意义(P<0.05);其中7个蛋白质在肱二头肌和小鱼际肌中的表达量是不同的.被鉴定的7个蛋白质可归类为代谢蛋白、分子伴侣蛋白、收缩性蛋白.结论该结果证实使用蛋白质组学分析骨骼肌萎缩相关蛋白质是可行和有效的,并首次提供了人类健康骨骼肌组织蛋白质2-D图谱.研究了失神经萎缩骨骼肌蛋白质组学的结果表明,细胞内蛋白质合成、防止自由基团的攻击、蛋白质错误折叠的修复以及细胞中蛋白质分选均与人类臂丛损伤后失神经骨骼肌萎缩的病程相关.Objective To investigate the changes in protein expression in human skeletal muscles undergoing denervation atrophy by using proteomics analysis and evaluate its clinical significance. Methods 9 patients of brachial plexus avulsion injuries were involved in the study. Atrophied hypothenar muscle and biceps muscle of the affected side and normal sternocleidomastoid muscle from the contralateral side were take. Two-dimensional electrophoresis and PD-Quest were applied for all muscle samples in order to comprehend protein expression regularity of muscles undergoing denervation atrophy. The peptide mass fingerprinting of difference protein was analyzed by matrixassisted laser desorption/ionization-time-of-flight （MALDI-TOF） mass spectrometer. The results of peptide mass fingerprinting were identified using the programs MS-Fit. Results （ 1 ） Technique platform for proteomics analysis for human skeletal muscles was preliminarily set up. Two-dimensional electrophoresis images for human normal muscles and denervation atrophy muscles were obtained. Manifest difference between normal muscles and denervation atrophy muscles was easily detected. （2）An average of 800 protein spots per gel were detected in denervation atrophy muscles,among which a total 32 spots were significantly altered between normal and atrophy muscles. Of this group, a total of 6 spots displayed increase in expression levels in the atrophy group as compared to normal （ P 〈 0.05）, while 24 proteinsdis played decrease in expression levels in atrophy over normal muscles （ P 〈 0.05 ）. （3） Proteins were identified by pepfide mass fingerprinting using MALDI-TOF. So far 7 proteins have been identified. Data on the identified spots were altered proteins in atrophy muscles. The identified proteins could be generally grouped together as metabolic proteins,chaperone proteins and contractile apparatus proteins. Conclusion The present work demonstrated the feasibility and validity of using proteomics analysis to search for skeletal muscle a

关 键 词：失神经萎缩 骨骼肌 蛋白质组学 肌细胞 自由基团 

分 类 号：R746[医药卫生—神经病学与精神病学]